Sample records for monoclonal antibody ki-67

  1. Ki-67 immunoreactivity in meningiomas--determination of the proliferative potential of meningiomas using the monoclonal antibody Ki-67

    DEFF Research Database (Denmark)

    Madsen, C; Schrøder, H D


    The proliferative potential of 66 human intracranial meningiomas (15 benign, 15 atypical, 15 recurrent, 13 bone-invasive, and 8 brain-invasive) was investigated by means of immunohisto-chemistry using the monoclonal antibody Ki-67. This antibody recognizes a nuclear antigen present in human cells...... during all active phases of the cell cycle, but absent in the resting phase. The purpose of this retrospective study was to estimate the Ki-67-labelling index (L.I.) and see if this index could help discriminate between the different groups of meningiomas. Our results demonstrated that L.I. could...

  2. Measurement of Ki-67 antigen in 159 pituitary adenomas using the MIB-1 monoclonal antibody

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    C.B. Pizarro


    Full Text Available Pituitary adenomas sometimes show rapid growth and recurrence, and about one third invade the structures surrounding the sella turcica. In an attempt to determine aggressive behavior at an early stage, we used the MIB-1 antibody to identify the Ki-67 antigen. The present study was designed to evaluate pituitary adenomatous tissue in terms of secretion and proliferation and to correlate the Ki-67 index with hormone phenotype and invasive behavior. Material from 159 patients submitted to one or more resections of pituitary adenomas was evaluated. Forty-two non-secretory adenomas and 43 adenomas immunoreactive for growth hormone, 19 for prolactin, 18 for growth hormone and prolactin, 16 for adrenocorticotropic hormone (ACTH, and 21 cases of plurihormonal/gonadotropin adenomas were detected by immunohistochemistry. The MIB-1 antibody was positive in 139 samples and the Ki-67 index ranged from 0.16 to 15.48% (mean = 1.22 ± 2.09%, with no significant difference between genders, age groups, or secretory and non-secretory status. The Ki-67 index was higher in ACTH-secreting adenomas. Invasive pituitary adenomas had a significantly higher Ki-67 index (2.01 ± 3.15% than macroadenomas with or without supra-sellar extension (1.12 ± 1.87%; P = 0.02. The index was not significantly different in the subgroup of adenomas with invasion of the cavernous sinus compared to groups with other types of invasion. We conclude that tumoral proliferative activity evaluated by the detection of the Ki-67 antigen is significantly higher in invasive than noninvasive adenomas, information which can be useful in therapeutic postoperative management since index cut-off values associated with aggressive behavior can be established.

  3. Hubungan Ekspresi Ki-67 dengan Grading Histopatologi Liposarkoma

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    Full Text Available Liposarcoma, a malignant tumor of mesenchymal origin, was one of the most common sarcomas of adults and occurs primarily in the fifth through seventh decades of life. The occurrence in children was rare. Liposarcoma were occur most frequently in the lower extremities (popliteal fossa and medial thigh, retroperitoneal, perirenal, mesenteric region and shoulder area. Retrospective study was conducted by reviewing the data from Pathology Anatomy Department Dr. Hasan Sadikin Hospital Bandung. There were 30 cases of liposarcoma, whereas lowgrade of liposarcoma 20 cases and high-grade of liposarcoma 10 cases. Research was done during February–May 2011. Immunohistochemical staining examination used monoclonal antibody Ki-67 as marker cell proliferation with the aim for looking Ki-67 expression and related with histopatholgy grade of liposarcoma. The tumor cells which showed Ki-67 expression were divided into very low (80% positive, whereas the intensity of Ki-67 staining was into weak, moderate and strong. There were significant differences between the Ki-67 expression and intensity level according to the histopathological types of liposarcoma (p<0.0001. In conclusions, Ki-67 expression increases with strong intensity of Ki-67 only in high-grade liposarcoma, so relates with progressivity of tumor.

  4. Utility of Ki-67 and p53 in distinguishing cervical intraepithelial neoplasia 3 from squamous cell carcinoma of the cervix. (United States)

    Tan, Geok Chin; Sharifah, Noor Akmal; Shiran, Mohd Sidik; Salwati, Shuib; Hatta, Ahmad Zailani; Paul-Ng, Hock Oon


    The differentiation between cervical intraepithelial neoplasia 3 (CIN 3) and early squamous cell carcinoma (SCC) of the cervix may be difficult in certain situations. Identification of invasion beyond the basement membrane is the gold standard for the diagnosis of the latter. The objective of this study was to determine whether the use of Ki-67 and p53 could help in solving the above dilemma. This was a retrospective study on 61 cases of cervical neoplasms comprising of 25 cases of CIN 3 and 36 SCC. All cases were evaluated by immunohistochemistry using Ki-67 and p53 monoclonal antibodies. Results showed that the differences of Ki-67 and p53 expression between CIN 3 and SCC were statistically significant. In conclusion, Ki-67 and p53 may serve as helpful adjuncts to routinely-stained histological sections in differentiating between CIN 3 and SCC.

  5. Increased Ki-67 immunoreactivity in the white matter in hemimegalencephaly. (United States)

    Munakata, Mitsutoshi; Watanabe, Mika; Otsuki, Taisuke; Itoh, Masayuki; Uematsu, Mitsugu; Saito, Yuko; Honda, Ryoko; Kure, Shigeo


    Hemimegalencephaly (HMG) is a developmental brain disorder characterized by an enlarged unilateral hemisphere with cortical malformation comprising abnormal hypertrophic cells. To address the proliferative status of HMG, Ki-67 immunoreactivity was investigated in HMG specimens obtained during epilepsy surgery. Nine HMG tissues were stained with a Ki-67 antibody and Ki-67 labeling index in the malformed cortex, and the underlying white matter was measured separately and compared with tissues from focal cortical dysplasias and normal brains from autopsy. In HMG tissues, Ki-67-positive cells were scattered in both the gray and white matter, with a significantly higher Ki-67 labeling index in the white matter compared with gray matter. No dysmorphic neuron or balloon cell was stained for Ki-67. As Ki-67 immunoreactivity overlapped with that of ionized calcium-binding adaptor protein-1, Ki-67-positive cells were identified as microglia. In HMG, microglia were activated and entered into a proliferative status with higher distribution in the white matter, implying an ongoing neuroinflammatory process involving the white matter. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  6. Prognostic Value of Proliferation Markers: Immunohistochemical Ki-67 Expression and Cytometric S-Phase Fraction of Women with Breast Cancer in Libya

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    Eramah Ermiah, Abdelbaset Buhmeida, Fathi Abdalla, Ben Romdhane Khaled, Nada Salem, Seppo Pyrhönen, Yrjö Collan


    Full Text Available Background: We evaluated the association of the immunohistochemical Ki-67 expression, and S-phase fraction with clinicopathological variables and patient outcome.Patients and methods: Histological samples from 100 primary Libyan breast carcinoma patients were retrospectively studied with monoclonal antibody to Ki-67. S-phase fraction was determined by DNA image cytometry.Results: The median Ki-67 percentage for all tumors was 27.5%, ranging from 1 to 80% and the median S-phase fraction (SPF was 11%, ranging from 0 to 62 %. Tumors with high Ki-67 expression were found in 76% of patients and with high SPF values in 56%. Ki-67 expression was more frequent in tumors with high SPF than low SPF. High Ki-67 and high SPF were associated with advanced stages, poor differentiation of tumors, positive lymph nodes, and distant metastasis. The Ki-67 was associated with hormone receptor negative tumors. The SPF was higher in young patients (<50 years than in older patients. In the overall population (median follow-up 49 months, patients with high Ki-67 and high SPF had shorter survival time and predicted recurrence than patients with low Ki-67 and low SPF. In a Cox multivariate analysis, high SPF (p= 0.007, hormonal status (p= 0.001 and clinical stage (p=0.005 were independent predictors of disease-specific survival. The Ki-67 (p=0.065 in borderline significance proved to be independent predictor of disease-free survival. The SPF showed more statistically significance with a high grade of malignancy and survival time than Ki-67.Conclusions: The SPF value is useful cell proliferation marker to assess tumor prognosis. These markers may reflect the aggressive behavior of Libyan breast cancer and predict of the recurrence. It is therefore important to take these markers into consideration to select a high risk subgroup of the patients for intensive treatment.

  7. A comparative study of Ki-67 antigen expression between luminal A and triple-negative subtypes of breast cancer. (United States)

    Borges, Umbelina Soares; Costa-Silva, Danylo Rafhael; da Silva-Sampaio, João Paulo; Escórcio-Dourado, Carla Solange; Conde, Airton Mendes; Campelo, Viriato; Gebrim, Luiz Henrique; da Silva, Benedito Borges; Lopes-Costa, Pedro Vitor


    Tumor biomarkers such as hormone receptors, HER-2 and Ki-67 are used routinely in clinical practice for classification of molecular subtypes of breast cancer. Cell proliferation evaluated by Ki-67 antigen expression is important to determine tumor aggressiveness. However, there is a paucity of studies comparing Ki-67 expression in an expressive number of cells among molecular subtypes of breast cancer, particularly among less and more aggressive tumors, such as luminal A and triple-negative, which have led us to the present study. The current study included invasive ductal carcinoma samples of 59 patients, which were divided into two groups: luminal A (n = 29) and triple-negative (n = 30). For immunohistochemical reaction, the samples were incubated with monoclonal anti-Ki-67 antibody (clone MIB1) and cells expressing Ki-67 protein were identified by dark brown staining of the nuclei, counting at least 600 cells per slide. The mean percentages of stained nuclei were analyzed by Student's t test (p < 0.05). The mean percentage of nuclei stained with anti-ki-67 was 10.14 and 77.22 in luminal A and triple-negative breast cancers, respectively (p < 0.0001). Our study showed a high cell proliferation of triple-negative breast cancer in comparison with luminal A, justifying its aggressiveness and poor clinical outcome.

  8. 探讨抗体标记物MC联合应用CK19,TPO,Galectin-3,KI-67在甲状腺乳头状病变中应用%Discuss application of antibody marker MC combining with CK19, TPO, Galectin-3, KI-67 in thyroid papillary lesions

    Institute of Scientific and Technical Information of China (English)

    朱春良; 帅剑波; 邵庆庆; 胡广明


    ABSTRACT:Objective through application of antibody marker MC combining with CK19, TPO, Galectin-3, KI-67 in expression status of thyroid papillary lesions, to find out the best combination of thyroid papillary carcinoma antibodies. Method Review and analyze expression of MC CK19, TPO, KI-67, Galectin-3 and MC of 96 cases papillary thyroid lesion with immunohistochemistry during Feb, 2012 and Feb, 2015. Result expression of the combination antibodies in 96 cases papillary thyroid lesion is as follows:64 case papillary thyroid carcinoma, and positive expression is respectively 10%(7/64), 96.9%(62/64 cases), 7.8%(5 cases weak positive), 100%(64/64), 62.5%(40/64). Thyroid papillary hyperplasia expression is as follows:9.4%(3/32 cases weak positive), 6.5%(2 cases positive), 12.5%(4/32) 22.5%(7 cases positive), 31.2%(10/32). Expression of KI-67, CK19, TPO, Galectin-3 and MC in two groups shows statistical significance (P<0.01). MC is a new marker for thyroid papillary carcinoma in recent years, and its expression in papillary thyroid carcinoma does not diffuse, and has little and weak expression in papillary hyperplasia. Conclusion MC combined with CK19, TPO, Galectin-3 and KI-67 in diagnosis of benign and malignant thyroid papillary lesion further provides reliable evidence, which is one of the better referential antibodies for diagnosis of thyroid papillary carcinoma.%目的:通过联合应用MC、CK19、TPO、Galectin-3、KI-67在甲状腺乳头状病变中表达情况,寻求找到甲状腺乳头状癌的抗体的最佳组合。方法回顾分析我院2012年2月至2015年2月期间96例甲状腺乳头状病变行免疫组化对MC、CK19、TPO、Galectin-3、KI-67表达。结果该组联合抗体在96例甲状腺乳头状病变中表达如下:甲状腺乳头状癌64例,阳性表达分别为:10%(7/64),96.9%(62/64例)、7.8%(5例弱阳性),100%(64/64)、62.5%(40/64)。甲状腺乳头状增生表达如下:9.4%(3/32例弱阳性),6.5%

  9. Immunohistochemicai study of Ki- 67 expression in unicystic Ameloblastoma and Dentigerous cyst

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    Eslami M.


    Full Text Available Statement of Problem: Differentiation of dentigerous cyst from unicystic ameloblastoma, discovering any initial ameloblastic changes in lining epithelium of dentigerous cyst at early stage, and differentiation between hyperplastic odontogenic epithelium in fibrous capsule of dentigerous cyst from ameloblastic proliferation, need to an accurate and reliable technique."nPurpose: The aim of this study was to determine and compare Ki-67 immunoreactivity in various locations of the epithelium of Dentigerous cyst and Unicystic Ameloblastoma."nMaterials and Methods: In this historical Cohort study, 15 cases of dentigerous cyst and 9 cases of unicystic ameloblastoma were selected. Immunohistochemistry staining was performed by M1B-1 (murine monoclonal antibody against Ki-67. The stained nucleous were counted in basal and suprabasal layer of lining epithelium of both lesions in 3000 epithelial cells. Finally, the percentage of positive cells (presented as labeling index was calculated, t- student test was used to analyze the related data."nResults: Ki-67 (LI in basal layer of Dentigerous cyst (2.59±1.66 and Unicystic Ameloblastoma (3.76±79 had no significant differences, but Ki-67 (LI in suprabasal layer of unicystic ameloblastoma (2.15±0.69 was significantly higher than dentigerous cyst (0.77±0.55 P=0.003."nThe difference between the average numbers of positive cells for Ki-67 (LI in these two lesions was statistically significant (P<0.05 and it was higher in Unicystic Ameloblastoma than Dentigerous cyst."nConclusion: Based on the findings of this study, it is suggested that Ki-67 (LI in suprabasal layer or throughout the epithelium can be considered as a useful marker for differential diagnosis between dentigerous cyst and unicystic ameloblastoma.

  10. Hubungan antara Imunoekspresi Ki-67 dan Risiko Agresivitas Tumor pada Gastrointestinal Stromal Tumor

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    Herry Yulianti


    Full Text Available Gastrointestinal stromal tumor (GIST is the most common mesenchymal tumor of the gastrointestinal tract, and arises from intestinal cells of Cajal localized in the muscular layer of the digestive tract, which functions as pacemaker cells in regulating intestinal motility. The incidence of GIST is about 3−5% of all soft tissue sarcomas. Gastrointestinal stromal tumor can occur along the gastrointestinal tract and predominantly in middle-aged and older persons, with a median age between 50 and 60 years. Histologically, there are three categories of GIST morphology such as spindle cells, epitheloid, and mixed type. A spesific marker of GIST is cluster of differentiation (CD117, which has good sensitivity and immunoreactive in 95% of GIST. The expression of Ki-67 correlates with proliferative activities and can be detected in G1, S, G2, and M phases of cell cycle but not in G0 phase. The aim of this study was to assessthe correlation between the risk of aggressive behaviors and proliferative activities as measured by Ki-67 in tumors confirmed as GIST by CD117. The method of this study was cross-sectional, performed on 29 cases of GIST from the Department of Pathology Anatomy Dr. Hasan Sadikin General Hospital/Faculty of Medicine Universitas Padjadjaran, Santo Borromeus Hospital, Immanuel Hospital, and Santosa Hospital between 2007−2012. A section from paraffin embedded tissue of 55 cases of GIST was stained with hematoxylin eosin for histological and immunohistochemical evaluations using monoclonal antibody CD117 to confirm the diagnosis of GIST. There were 29 positive cases for CD117. Further staining was performed using monoclonal antibody Ki-67. The categorized positive cells of immunoexpression of CD117 showed brown particles inside cytoplasma and the immunoexpression of Ki-67 was assessed by identification of nuclear brown staining of neoplastic cells. The result showed that there were significant correlations between the risk of tumor

  11. Study of Ki67 and CD10 expression as predictive factors of recurrence of ameloblastoma. (United States)

    Ahlem, B; Wided, A; Amani, L; Nadia, Z; Amira, A; Faten, F


    Ameloblastoma is a rare, benign, purely epithelial odontogenic tumour, characterized by a high potential for local invasion and recurrence. To study the epidemiological and histological characteristics of ameloblastoma. To study Ki67 and CD10 immunostaining in ameloblastoma and to investigate a possible correlation between these two markers and recurrence of this tumour. An immunohistochemical study using Ki67 and CD10 monoclonal antibodies was performed on 37 paraffin blocks obtained from the Charles-Nicolle hospital pathology department in Tunis over a 9-year period (2004-2012). Statistical analysis was performed with Statistical Package for Social Sciences (SPSS) software version 15.1. This series of 37 cases comprised 21 males and 16 females (sex ratio: 1.3) with a mean age of 39 years (range: 7 to 70 years), corresponding to 36 cases of intraosseous ameloblastoma and one case of gingival ameloblastoma. Thirty-two cases were polycystic and 5 cases were unicystic. Eighteen cases of local recurrence were observed. No correlation was demonstrated between recurrence and the various clinical and histological parameters and treatment modalities. However, a significant correlation was demonstrated between recurrence and Ki67 and CD10 expression (P=0.000 and 0.002, respectively). The Ki67 proliferation index and stromal CD10 expression can be considered to be predictive factors of ameloblastoma recurrence. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  12. EGFR, CD10 and proliferation marker Ki67 expression in ameloblastoma: possible role in local recurrence

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    Abdel-Aziz Azza


    Full Text Available Abstract Background Ameloblastoma is an odontogenic neoplasm characterized by local invasiveness and tendency towards recurrence. Aims Studying the role played by EGFR, CD10 and Ki67 in the recurrence of ameloblastoma. Methods This study was carried out on 22 retrospective cases of mandibular ameloblastoma from the period from Jan 2002 to Jan 2008 with follow up period until Jan 2011 (3 to 8 years follow up peroid. Archival materials were obtained from pathology department, Mansoura university. Paraffin sections of tumor tissue from all cases were submitted for routine H&E stains and immunohistochemistry using EGFR, CD10 and Ki67 monoclonal antibodies. Statistical analysis using of clinical data for all patients, tumor type, EGFR, CD10 and Ki67 expression in relation to recurrence were evaluated. Results Among the 22 cases, 10 cases were males and 12 were females with sex ratio 1:1.2. Age ranged from 34 to 59 years old with a mean age 44.18 year. Five cases showed local recurrence within studied period and proved by biopsy. No statistically significant relation was found between local recurrence and patient age, tumor size, tumor type, EGFR expression. There was a significant relation between CD10 expression as well as Ki67 labelling index and recurrence (P value = 0.003, 0.000 respectively. Conclusion Evaluation of CD10 and Ki67 status together with conventional histological evaluation can help in providing more information about the biologic behavior of the tumor, while EGFR could be a target of an expanding class of anticancer therapies. Since ameloblastomas are EGFR-positive tumors, anti-EGFR agents could be considered to reduce the size of large tumors and to treat unresectable tumors that are in close proximity to vital structures. Virtual Slides The virtual slide(s for this article can be found here:

  13. Cervical intraepithelial neoplasia in pregnancy: Interference of pregnancy status with p16 and Ki-67 protein expression (United States)

    Ciavattini, Andrea; Sopracordevole, Francesco; Di Giuseppe, Jacopo; Moriconi, Lorenzo; Lucarini, Guendalina; Mancioli, Francesca; Zizzi, Antonio; Goteri, Gaia


    To date, there are evidence-based guidelines available for cervical dysplasia diagnosed in pregnancy. Certain functional biomarkers have proven useful in the prediction of regressing and non-regressing cervical intraepithelial neoplasia (CIN) lesions in non-pregnant women. In the present study, Ki-67 and p16 immunostaining were evaluated in different grades of CIN lesions diagnosed in pregnant or non-pregnant women with the aim to identify any differences in order to better understand the behavior of CIN in pregnancy. The current retrospective case-control study included 17 pregnant patients that conceived naturally with first-time onset of CIN occurring at no later than 16 gestational weeks. The control group included 17 non-pregnant patients matched for age, parity and number of previous sexual partners. Exclusion criteria included previous cervical treatment, immunocompromised status, chronic hepatitis B and/or C and cigarette smoking. p16 and Ki-67 protein expression were respectively detected using the CINtec Histology kit and monoclonal antibodies against Ki-67. p16 and Ki-67 staining were analyzed using a classification system based on the distribution of positivity on a semi-quantitative three point-scale. p16 and Ki-67 immune reactivity correlated positively with the grade of epithelial dysplasia in the total cohort of pregnant and non-pregnant patients; expression increased linearly from CIN1 to CIN3. Furthermore, the association between p16 immunostaining and CIN grade was significant in non-pregnant patients but not in pregnant patients. In pregnant patients, positivity for Ki-67 was less intense than in non-pregnant patients. These results appear to suggest that pregnancy status interferes with the expression of cellular proteins involved in cell-cycle regulation and the carcinogenic process induced by high-risk human papilloma virus, exhibiting increased variability in their staining. PMID:28123559

  14. Therapeutic Recombinant Monoclonal Antibodies (United States)

    Bakhtiar, Ray


    During the last two decades, the rapid growth of biotechnology-derived techniques has led to a myriad of therapeutic recombinant monoclonal antibodies with significant clinical benefits. Recombinant monoclonal antibodies can be obtained from a number of natural sources such as animal cell cultures using recombinant DNA engineering. In contrast to…

  15. Therapeutic Recombinant Monoclonal Antibodies (United States)

    Bakhtiar, Ray


    During the last two decades, the rapid growth of biotechnology-derived techniques has led to a myriad of therapeutic recombinant monoclonal antibodies with significant clinical benefits. Recombinant monoclonal antibodies can be obtained from a number of natural sources such as animal cell cultures using recombinant DNA engineering. In contrast to…

  16. Effects of low-dose tamoxifen on breast cancer biomarkers Ki-67, estrogen and progesterone receptors. (United States)

    de Sousa, Juarez Antônio; Facina, Gil; da Silva, Benedito Borges; Gebrim, Luiz Henrique


    Breast carcinoma is the most common malignancy among women and it has a major impact on mortality. Studies of primary chemoprevention with tamoxifen have generated high expectations and considerable success rates. The efficacy of lower doses of tamoxifen is similar to that seen with a standard dose of the drug, and there has been a reduction in healthcare costs and side effects. The immune reaction to monoclonal antibody Ki-67 (MIB-1) and the expression of estrogen receptors (1D5) and progesterone receptors (PgR 636) in breast carcinoma were studied in patients treated with 10 mg of tamoxifen for a period of 14 days. A prospective randomized clinical trial was conducted with 38 patients divided into two groups: Group A: N = 20 (control group--without medication) and Group B: N = 18 (tamoxifen/10 mg/day for 14 days). All patients signed an informed consent term previously approved by both institutions. Patients underwent incisional biopsy before treatment and 14 days later a tumor tissue sample was obtained during surgical treatment. Positivity was quantitatively assessed, counting at least 1.000 cells per slide. For statistical data analysis, a Wilcoxon non-parametric test was used, and alpha was set at 5%. Both groups (A and B) were considered homogeneous regarding control variables. In Group A (control), there was no statistically significant reduction in Ki-67 (MIB-1) (p = 0.627), estrogen receptor (1D5) (p = 0.296) and progesterone receptor positivity (PgR 636) (p = 0.381). In Group B (tamoxifen 10 mg/day), the mean percentage of nuclei stained by Ki-67 (MIB-1) was 24.69% before and 10.43% after tamoxifen treatment. Mean percentage of nuclei stained by estrogen receptor (1D5) was 59.53% before and 25.99% after tamoxifen treatment. Mean percentage of nuclei stained by progesterone receptor (PgR 636), was 59.34 before and 29.59% after tamoxifen treatment. A statistically significant reduction was found with the three markers (p Tamoxifen significantly reduced

  17. Monoclonal antibody "gold rush". (United States)

    Maggon, Krishan


    The market, sales and regulatory approval of new human medicines, during the past few years, indicates increasing number and share of new biologics and emergence of new multibillion dollar molecules. The global sale of monoclonal antibodies in 2006 were $20.6 billion. Remicade had annual sales gain of $1 billion during the past 3 years and five brands had similar increase in 2006. Rituxan with 2006 sales of $4.7 billion was the best selling monoclonal antibody and biological product and the 6th among the top selling medicinal brand. It may be the first biologic and monoclonal antibody to reach $10 billion annual sales in the near future. The strong demand from cancer and arthritis patients has surpassed almost all commercial market research reports and sales forecast. Seven monoclonal antibody brands in 2006 had sales exceeding $1 billion. Humanized or fully human monoclonal antibodies with low immunogenicity, enhanced antigen binding and reduced cellular toxicity provide better clinical efficacy. The higher technical and clinical success rate, overcoming of technical hurdles in large scale manufacturing, low cost of market entry and IND filing, use of fully human and humanized monoclonal antibodies has attracted funds and resources towards R&D. Review of industry research pipeline and sales data during the past 3 years indicate a real paradigm shift in industrial R&D from pharmaceutical to biologics and monoclonal antibodies. The antibody bandwagon has been joined by 200 companies with hundreds of new projects and targets and has attracted billions of dollars in R&D investment, acquisitions and licensing deals leading to the current Monoclonal Antibody Gold Rush.

  18. The region-of-interest size impacts on Ki67 quantification by computer-assisted image analysis in breast cancer. (United States)

    Christgen, Matthias; von Ahsen, Sabrina; Christgen, Henriette; Länger, Florian; Kreipe, Hans


    Therapeutic decision-making in breast cancer depends on histopathologic biomarkers and is influenced by the Ki67 proliferation index. Computer-assisted image analysis (CAIA) promises to improve Ki67 quantification. Several commercial applications have been developed for semiautomated CAIA-based Ki67 quantification, many of which rely on measurements in user-defined regions of interest (ROIs). Because of intratumoral proliferative heterogeneity, definition of the ROI is an important step in the analytical procedure. This study explores the ROI size impacts on Ki67 quantification. Whole-slide sections of 100 breast cancers were immunostained with the anti-Ki67 antibody 30-9 and were analyzed on the iScan Coreo digital pathology platform using a Food and Drug Administration-cleared Ki67 quantification software version v5.3 (Virtuoso; Ventana, Tucson, TX). For each case, the Ki67 labeling index (LI) was determined in multiple ROIs of gradually increasing size centered around a high-proliferation area. The spatial Ki67 decline was modeled with nonlinear regression. Depending on the ROI size, the median Ki67 LI varied between 55% and 15%. The proportion of tumors classified as Ki67 low according to the St Gallen 2013/2015 cutoff increased from 2% to 56%, as the ROI size increased from 50 to 10,000 cells captured. The interrater reliability of conventional Ki67 assessment versus CAIA-based Ki67 quantification was also dependent on the ROI size and varied between slight and almost perfect agreement (Cohen κ = 0.06-0.85). In conclusion, the ROI size is a critically important parameter for semiautomated Ki67 quantification by CAIA. Ki67 LIs determined on platforms like iScan Coreo/Virtuoso require an ROI size adjustment, for which we offer a downloadable data transformation tool.

  19. Heterogeneity of monoclonal antibodies. (United States)

    Liu, Hongcheng; Gaza-Bulseco, Georgeen; Faldu, Dinesh; Chumsae, Chris; Sun, Joanne


    Heterogeneity of monoclonal antibodies is common due to the various modifications introduced over the lifespan of the molecules from the point of synthesis to the point of complete clearance from the subjects. The vast number of modifications presents great challenge to the thorough characterization of the molecules. This article reviews the current knowledge of enzymatic and nonenzymatic modifications of monoclonal antibodies including the common ones such as incomplete disulfide bond formation, glycosylation, N-terminal pyroglutamine cyclization, C-terminal lysine processing, deamidation, isomerization, and oxidation, and less common ones such as modification of the N-terminal amino acids by maleuric acid and amidation of the C-terminal amino acid. In addition, noncovalent associations with other molecules, conformational diversity and aggregation of monoclonal antibodies are also discussed. Through a complete understanding of the heterogeneity of monoclonal antibodies, strategies can be employed to better identify the potential modifications and thoroughly characterize the molecules.

  20. Prognostic significance of Ki-67 in salivary gland carcinomas

    DEFF Research Database (Denmark)

    Larsen, Stine Rosenkilde; Bjørndal, Kristine; Godballe, Christian;


    J Oral Pathol Med (2012) Background:  Salivary gland carcinomas are a heterogeneous group of tumors with varying malignant potential. In this study, we evaluated the proliferative marker Ki-67 in salivary gland carcinomas and related the Ki-67 index to clinical data. Methods:  A total of 176...... salivary gland carcinomas of 13 different subtypes were stained immunohistochemically for Ki-67. The number of Ki-67 positive cells was counted and the Ki-67 index was calculated as the percentage of positive tumor cells. Results:  The Ki-67 median value was 26 (range 1-99). The median follow-up time was 6...

  1. Comparative immunoexpression of ICAM-1, TGF-?1 and ki-67 in periapical and residual cysts


    Martins, Rafaela; Armada, Luciana; dos-Santos, Teresa-Cristina-Ribeiro-Bartholomeu; Pires, Fábio-Ramôa


    Background This study compared the immunohistochemical expression of ki-67, transforming growth factor beta 1 (TGF-β1) and intercellular adhesion molecule-1 (ICAM-1) in inflammatory periapical cysts and residual cysts. Material and Methods The study sample was composed by 25 periapical cysts and 25 residual cysts and immunohistochemical reactions were carried out using antibodies directed against ICAM-1, TGF-β1 and ki-67. Clinical, radiological, gross, histological and immunohistochemical dat...

  2. Expression of p53, Ki-67, and CD31 proteins in endometrial polyps of postmenopausal women treated with tamoxifen. (United States)

    Miranda, Sergimar P; Traiman, Paulo; Cândido, Eduardo B; Lages, Elisa L; Freitas, Gustavo F; Lamaita, Rívia Mara; Vidigal, Paula V T; da Silva Filho, Agnaldo Lopes


    This study was undertaken to investigate the expression of p53, Ki-67, and CD31 proteins in endometrial polyps of postmenopausal women treated with tamoxifen (TAM). Postmenopausal women with endometrial polyps treated with TAM (n = 20), postmenopausal women with endometrial polyps without hormone use (n = 20), postmenopausal women with atrophic endometrium (n = 20), and postmenopausal women with endometrial adenocarcinoma (n = 20) were prospectively investigated. Tissue samples were immunohistochemically evaluated by monoclonal antibodies for p53, Ki-67, and CD31. The data were analyzed using the Student t test, analysis of variance, and χ2 to evaluate significant differences between the groups. The level of significance was set at P < 0.05. There was no difference in the expression of p53 between the groups (P = 0.067). The expression of Ki-67 was higher in the polyp samples from TAM-treated women compared with those from the women using no hormone (P = 0.0047) and those from the women with atrophic endometrium (P = 0.008). Samples from the women with endometrial cancer was associated with higher Ki-67 expression compared with the polyp samples from TAM-treated women (P = 0.004). The expression of CD31 was higher in the polyp samples of TAM-treated women compared with that of the samples from the women with atrophic endometrium (P < 0.001) and similar to the polyp samples from the women using no hormone (P = 0.319) and to the samples from the women with endometrial cancer (P = 0.418). The use of TAM in postmenopausal women might be associated with increased cellular proliferation in endometrial polyps without interfering angiogenesis or inactivation of tumor suppressor proteins.


    Institute of Scientific and Technical Information of China (English)

    黄高升; FellerAC; LennertK


    Proliferative activity in 106 cases of non-Hodgkin's lymphoma was estimated using the monoclonal antibody Ki67 and by countin the number of mitotic figures.The percentage of Ki67-positive cells was compared with the median mumber of mitotic figures per square millimeter.Both Ki67 positivity and the number of mitotic figures were found to be greater in high grade than in low grade lymphomas,although there was an overlap between the two grades of malignancy.A close correlation was found between the number of mitotic figured and the percentage of Ki67-positive cells not only in all lymphoma types taken together (rs=0.834,P<0.001),but also in B-cell lymphoma(rs=0.818,P<0.001)and T-cell lymphoma(rs=0.764,P<0.001)taken seprately.Thus,both methods are useful for the estimation of proliferative activity,but each method has is advantages and disadvan-tages.

  4. Monoclonal antibodies in myeloma

    DEFF Research Database (Denmark)

    Sondergeld, P.; van de Donk, N. W. C. J.; Richardson, P. G.;


    The development of monoclonal antibodies (mAbs) for the treatment of disease goes back to the vision of Paul Ehrlich in the late 19th century; however, the first successful treatment with a mAb was not until 1982, in a lymphoma patient. In multiple myeloma, mAbs are a very recent and exciting add...

  5. The expression and significance of p53 protein and Ki-67 protein in pterygium

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    Ljubojević Vesna


    Full Text Available Background/Aim. Pterygium is considered to be a degenerative disease of the conjunctiva, however, the presence of tumor markers in pterygium reinforces the hypothesis that this lesion is similar to tumor. Inactivation of p53 function removes an obstacle to increased proliferation. Factors affecting the prevalence of p53 expression in pterygium deserve investigation. The aim of the study was to investigate the expression of p53 and Ki-67 proteins in pterygium and normal conjunctiva, the effects of gender and age on p53 expression, and the relationship between the expression of p53 and Ki-67 proteins. Methods. A total of 34 samples of pterygium and 34 samples of the normal conjunctiva were analyzed. The samples were studied by immunohistochemistry using antibodies against p53 and Ki-67. Results. Totally 15 (44% samples of pterygia were p53 positive. Correlations between the expression of p53 protein and sex, and age were not established. The number of Ki-67 positive cells in pterygium (9.74% was significantly higher than the number of Ki-67 positive cells in the normal conjunctiva (1.74%, (p = 0.001. Between the expression of p53 protein and Ki-67 protein in pterygium there was a significant positive correlation (p = 0.000. Conclusion. The prevalence of p53 positive samples of pterygium was 44%. The influence of sex and age on p53 protein expression in pterygium was not found. The increased proliferative acivity was present in the epithelium of pterygium. The expression of Ki-67 protein is associated with the expression of p53 protein in pterygium. The findings of our study support the thesis of pterygium as tissue growth disorder.

  6. Expression and significance of E-cadherin and Ki-67 in non-special type of invasive ductal carcinoma of breast%E-钙黏蛋白和Ki-67在非特殊型乳腺浸润性导管癌中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    李玉红; 贾爱华; 鲁庆阳; 任玉波


    Objective To investigate the role of E-cadherin and antigen identified by monoclonal antibody Ki-67 in the occurrence and development of non-special type of invasive ductal carcinoma of breast. Methods The expression of E-cadherin and Ki-67 in 100 non-special type of invasive ductal carcinoma ( breast cancer group) and 20 breast fibroadenoma tissues ( fibroadenoma group) were detected by immunohistochemistry, and the relationship between these two antibodies and the clinicopathological features of non-special type of invasive ductal carcinoma were analyzed. Results The breast cancer group (67%) was lower than than the fibroadenoma group ( 100% ) in E-csdherin positive expression rate, and was ( 59% ) higher than the fibroadenoma group( 25% ) in Ki-67 positive expression rate, both P < 0.05; the expression of E-cadherin and Ki-67 were closely related to non-special type of invasive ductal carcinoma of breast in it's lymph node metastasis, TNM stage and histological grade, but not in patients' age and tumor diameter. Conclusions Reduced expression of E-cadherin and elevated expression of Ki-67 were closely related to non-special type of invasive ductal carcinoma of breast in it's occurrence and development, and detection of these two antibodies is helpful for determining the degree of malignancy and prognosis of non-special type of invasive ductal carcinoma of breast.%目的 探讨E-钙黏蛋白和增殖指数抗原Ki-67在非特殊型乳腺浸润性导管癌发生、发展中的作用.方法 采用免疫组化法检测E-钙黏蛋白和Ki-67在100份非特殊型乳腺浸润性导管癌组织(乳腺癌组)及20份乳腺纤维腺瘤组织(纤维腺瘤组)中的表达,并分析两者与非特殊型乳腺浸润性导管癌临床病理特征的关系.结果 乳腺癌组E-钙黏蛋白的阳性率(67%)明显低于纤维腺瘤组(100%),Ki-67的阳性率(59%)高于纤维腺瘤组(25%),P均<0.05;E-钙黏蛋白和Ki-67表达与非特殊型乳腺浸润性导管癌

  7. Phospho-Akt immunoreactivity in prostate cancer: relationship to disease severity and outcome, Ki67 and phosphorylated EGFR expression.

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    Peter Hammarsten

    Full Text Available BACKGROUND: In the present study, we have investigated the prognostic usefulness of phosphorylated Akt immunoreactivity (pAkt-IR in prostate cancer using a well-characterised tissue microarray from men who had undergone transurethral resection due to lower urinary tract symptoms. METHODOLOGY/PRINCIPAL FINDINGS: pAkt-IR in prostate epithelial and tumour cells was assessed using a monoclonal anti-pAkt (Ser(473 antibody. Immunoreactive intensity was determined for 282 (tumour and 240 (non-malignant tissue cases. Tumour pAkt-IR scores correlated with Gleason score, tumour Ki67-IR (a marker of cell proliferation and tumour phosphorylated epidermal growth factor receptor (pEGFR-IR. For cases followed with expectancy, a high tumour pAkt-IR was associated with a poor disease-specific survival, and the prognostic information provided by this biomarker was additive to that provided by either (but not both tumour pEFGR-IR or Ki67-IR. Upon division of the cases with respect to their Gleason scores, the prognostic value of pAkt-IR was seen for patients with Gleason score 8-10, but not for patients with Gleason score 6-7. CONCLUSIONS/SIGNIFICANCE: Tumour pAkt-IR is associated with both disease severity and disease-specific survival. However, its clinical use as a biomarker is limited, since it does not provide prognostic information in patients with Gleason scores 6-7.

  8. Interobserver Variability of Ki-67 Measurement in Breast Cancer

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    Yul Ri Chung


    Full Text Available Background: As measurement of Ki-67 proliferation index is an important part of breast cancer diagnostics, we conducted a multicenter study to examine the degree of concordance in Ki-67 counting and to find factors that lead to its variability. Methods: Thirty observers from thirty different institutions reviewed Ki-67–stained slides of 20 different breast cancers on whole sections and tissue microarray (TMA by online system. Ten of the 20 breast cancers had hot spots of Ki-67 expression. Each observer scored Ki-67 in two different ways: direct counting (average vs. hot spot method and categorical estimation. Intraclass correlation coefficient (ICC of Ki-67 index was calculated for comparative analysis. Results: For direct counting, ICC of TMA was slightly higher than that of whole sections using average method (0.895 vs 0.858. The ICC of tumors with hot spots was lower than that of tumors without (0.736 vs 0.874. In tumors with hot spots, observers took an additional counting from the hot spot; the ICC of whole sections using hot spot method was still lower than that of TMA (0.737 vs 0.895. In categorical estimation, Ki-67 index showed a wide distribution in some cases. Nevertheless, in tumors with hot spots, the range of distribution in Ki-67 categories was decreased with hot spot method and in TMA platform. Conclusions: Interobserver variability of Ki-67 index for direct counting and categorical estimation was relatively high. Tumors with hot spots showed greater interobserver variability as opposed to those without, and restricting the measurement area yielded lower interobserver variability.

  9. Analysis of the relationship between cancer procoagulant activity and PCNA and Ki-67 expression in cases of common and cellular uterine leiomyomas. (United States)

    Szajda, Slawomir D; Jóźwik, Marcin; Sulkowska, Mariola; Chabielska, Ewa; Sulkowski, Stanislaw; Jóźwik, Maciej


    Histological subtypes of uterine leiomyomas may substantially differ in their cellular biology, including the intensity of synthesis of cancer markers and expression of cell proliferation markers. The present investigation aimed to determine the activity of cancer procoagulant (CP) in subtypes of leiomyomas, including cellular leiomyomas, and to verify whether these activities correlate with immunoexpression of cell proliferation markers: the proliferating cell nuclear antigen (PCNA) and Ki-67. Preoperative peripheral venous blood and postoperative tissue material were obtained from 24 women operated on in a tertiary referral academic department. The activity of CP in serum was measured with the use of a coagulative method according to Gordon and Benson, and in tissue homogenates with the use of a spectrophotometric method according to Colucci et al. The control serum values were obtained from 20 healthy women without any gynecological disease, and the control solid tissue values from histologically confirmed postoperative normal reproductive tissues obtained from six patients. PCNA and Ki-67 expression were determined immunohistochemically using monoclonal antibodies. Both the tissue and serum activity for CP was considerably higher for common leiomyomas and cellular leiomyomas than for control tissues, but did not differ significantly between the leiomyoma subtypes. Intratumor CP activity significantly correlated with PCNA expression but not with Ki-67 expression. Cellular leiomyomas do not differ substantially in the serum and intratumor CP activity from common leiomyomas. There is a relationship of intratumor CP activity with PCNA expression, a finding which requires further investigation.

  10. Perichromosomal protein Ki67 supports mitotic chromosome architecture. (United States)

    Takagi, Masatoshi; Natsume, Toyoaki; Kanemaki, Masato T; Imamoto, Naoko


    Although the condensin complexes and topoisomerase IIα (TopoIIα) are the central players in mitotic chromosome formation, they are insufficient for its completion, and additional factors involved in the process have been extensively sought. In this study, we examined the possibility that Ki67, a perichromosomal protein widely used as a cell proliferation marker, is one such factor. Using a combination of auxin-inducible degron and CRISPR-Cas9-based gene editing technologies, we generated a human HCT116 cell line in which Ki67 is rapidly depleted in a few hours. The removal of Ki67 before mitotic entry did not impact the early mitotic chromosome assembly observed in prophase but subsequently resulted in the formation of misshapen mitotic chromosomes. When Ki67 was removed after mitotic entry, preassembled rod-shaped mitotic chromosomes became disorganized. In addition, we show that Ki67 and TopoIIα are reciprocally coimmunoprecipitated from mitotic cell extracts. These observations indicate that Ki67 aids the finalization of mitotic chromosome formation and helps maintain rod-shaped chromosome architecture, likely in collaboration with TopoIIα. Together, these findings represent a new model in which mitotic chromosome architecture is supported both internally and externally. © 2016 Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd.

  11. Ki-67 expression in anal intraepithelial neoplasia in AIDS

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    Edenilson Eduardo Calore


    Full Text Available CONTEXT: AIDS is one of the most important risk factors for progression and recurrence of anogenital condyloma. In a previous work, we observed that patients with warts and high-grade AIN (HAIN had recurrences more frequently than did patients with warts without AIN. The mechanisms of this increased incidence of high-grade lesions in AIDS are not known. OBJECTIVE: We studied the expression of the proliferative marker Ki-67 by immunohistochemical methods, in specimens of anal condyloma from HIV+ patients to clarify whether its expression can be associated to the grade of AIN. DESIGN: A retrospective study of hiltological specimens. SETTING: University referral unit. SAMPLE: 34 patients were divided into two groups: (1 condylomas with low grade AIN (LAIN, with 25 patients; and (2 condylomas with HAIN, with 9 patients. In this latter group we examined two areas: 2A (HAIN area and 2B (LAIN area. MAIN MEASUREMENTS: The immunohistochemical reaction for Ki-67 was done on histological sections. Slices were lightly stained with hematoxylin, to help us in Ki-67 positive cell counting. The percentage of Ki-67 marked nuclei was calculated. We applied one-way variance analysis for statistics. RESULTS: The mean number of Ki-67 positive cells in group 1 was 19.68 ± 10.99; in group 2 (area A it was 46.73 ± 10.409; and in area B it was 36.43 ± 14.731. There were statistical differences between groups 1 and 2A and between groups 1 and 2B. Ki-67 positive cells predominated in the lower layer in LAIN. Positive Ki-67 cells were found in all layers in group 2A, and in group 2B they predominated in the two lower or in all layers of the epithelium. CONCLUSIONS: Our results suggest that LAIN areas (using routine staining techniques in HAIN can have a biological behavior more similar to HAIN.

  12. Prothymosin-alpha and Ki-67 expression in pituitary adenomas

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    Iga Wierzbicka-Tutka


    Full Text Available Introduction: Prothymosin alpha (PTMA, a nuclear oncoprotein involved in cell cycle regulation, is used as a prognostic marker in many cancers. The histopathology of pituitary carcinomas and locally invasive adenomas is indistinguishable from that of benign tumors. A new marker is needed to differentiate these lesions. We evaluated PTMA in pituitary adenomas to determine its usefulness as a prognostic factor of tumor proliferation.Material/Methods: We conducted a retrospective analysis of a group of 27 patients, including 15 females (56% and 12 males (44% with a mean age of 58.6±12 years, who underwent pituitary tumor surgery between 2003 and 2012. The Ki-67 and PTMA-nuclear (PTMA-n and PTMA-cytoplasmic (PTMA-c indices were determined by immunohistochemical staining. We studied histopathological features, clinical symptoms, and magnetic resonance imaging or computed tomography performed before surgery and one year following surgery to evaluate tumor size and progression.Results: The expression of Ki-67 was revealed in 77.8% of adenomas, PTMA-n in 81.5% and PTMA-c in 92.6%. The mean value of the Ki-67 index was 1.8%, PTMA-n was 1.84%, and PTMA-c was 35.6%. There was a significant positive correlation between Ki-67 and PTMA-n (p=0.009. We did not find any correlation between Ki-67, PTMA-c, and tumor progression. PTMA-n was found to be correlated with tumor size (p=0.045 and was higher in the case of gonadotropinomas (p=0.026.Conclusions: The positive nuclear expression of Ki-67 and PTMA was observed in the majority of pituitary adenomas. Neither the expression of Ki-67 nor that of PTMA-c was related to tumor recurrence or local invasion.

  13. Fragmentation of monoclonal antibodies (United States)

    Vlasak, Josef


    Fragmentation is a degradation pathway ubiquitously observed in proteins despite the remarkable stability of peptide bond; proteins differ only by how much and where cleavage occurs. The goal of this review is to summarize reports regarding the non-enzymatic fragmentation of the peptide backbone of monoclonal antibodies (mAbs). The sites in the polypeptide chain susceptible to fragmentation are determined by a multitude of factors. Insights are provided on the intimate chemical mechanisms that can make some bonds prone to cleavage due to the presence of specific side-chains. In addition to primary structure, the secondary, tertiary and quaternary structures have a significant impact in modulating the distribution of cleavage sites by altering local flexibility, accessibility to solvent or bringing in close proximity side chains that are remote in sequence. This review focuses on cleavage sites observed in the constant regions of mAbs, with special emphasis on hinge fragmentation. The mechanisms responsible for backbone cleavage are strongly dependent on pH and can be catalyzed by metals or radicals. The distribution of cleavage sites are different under acidic compared to basic conditions, with fragmentation rates exhibiting a minimum in the pH range 5–6; therefore, the overall fragmentation pattern observed for a mAb is a complex result of structural and solvent conditions. A critical review of the techniques used to monitor fragmentation is also presented; usually a compromise has to be made between a highly sensitive method with good fragment separation and the capability to identify the cleavage site. The effect of fragmentation on the function of a mAb must be evaluated on a case-by-case basis depending on whether cleavage sites are observed in the variable or constant regions, and on the mechanism of action of the molecule. PMID:21487244

  14. An international study to increase concordance in Ki67 scoring

    DEFF Research Database (Denmark)

    Polley, Mei-Yin C; Leung, Samuel C Y; Gao, Dongxia


    Although an important biomarker in breast cancer, Ki67 lacks scoring standardization, which has limited its clinical use. Our previous study found variability when laboratories used their own scoring methods on centrally stained tissue microarray slides. In this current study, 16 laboratories from...... these data are potentially encouraging, suggesting that it may be possible to standardize scoring of Ki67 among pathology laboratories, clinically important discrepancies persist. Before this biomarker could be recommended for clinical use, future research will need to extend this approach to biopsies...

  15. Automated detection of dual p16/Ki67 nuclear immunoreactivity in liquid-based Pap tests for improved cervical cancer risk stratification. (United States)

    Gertych, Arkadiusz; Joseph, Anika O; Walts, Ann E; Bose, Shikha


    The Papanicolau (Pap) test is a routine cytological procedure for early detection of dysplastic lesions in cervical epithelium. A reliable screening method is crucial for triage of women at risk; however manual screening and interpretation are associated with relatively low sensitivity and substantial interobserver diagnostic variability. P16 and Ki67 biomarkers have been recently proposed as adjunctive tools in the diagnosis of high-risk human papillomavirus (hrHPV) associated dysplasias to supplement the morphological characteristics of cells by additional colorimetric features. In this study, an automated technique for the evaluation of dual p16/Ki67 immunoreactivity in cervical cell nuclei is introduced. Smears stained with p16 and Ki67 antibodies were digitized, and analyzed by algorithms we developed. Gradient-based radial symmetry operator and adaptive processing of symmetry image were employed to obtain the nuclear mask. This step was followed by the extraction of features including pixel data and immunoreactivity signature from each nucleus. The features were analyzed by two support vector machine classifiers to assign a nucleus into one of four types of immunoreactivity: p16 positive (p16(+)/Ki67(-)), Ki67 positive (p16(-)/Ki67(+)), dual p16/Ki67 positive (p16(+)/Ki67(+)) and negative (p16(-)/Ki67(-)), respectively. Results obtained by our method correlated well with readings by two cytopathologists (n = 18,068 cells); p16(+)/Ki67(+) nuclei were classified with respective precisions of 77.1% and 82.6%. Specificity in identification of p16(-)/Ki67(-) nuclei was better than 99.5%, and the sensitivity in detection of all immunopositive nuclei was 86.3 and 89.4%, respectively. We found that the quantitative characterization of immunoreactivity provided by the additional highlighting of classified nuclei can positively impact the efficacy and screening outcome of the Pap test.

  16. Uses of monoclonal antibody 8H9 (United States)

    Cheung, Nai-Kong V.


    This invention provides a composition comprising an effective amount of monoclonal antibody 8H9 or a derivative thereof and a suitable carrier. This invention provides a pharmaceutical composition comprising an effective amount of monoclonal antibody 8H9 or a derivative thereof and a pharmaceutically acceptable carrier. This invention also provides an antibody other than the monoclonal antibody 8H9 comprising the complementary determining regions of monoclonal antibody 8H9 or a derivative thereof, capable of binding to the same antigen as the monoclonal antibody 8H9. This invention provides a substance capable of competitively inhibiting the binding of monoclonal antibody 8H9. This invention also provides an isolated scFv of monoclonal antibody 8H9 or a derivative thereof. This invention also provides the 8H9 antigen. This invention also provides different uses of the monoclonal antibody 8H9 or its derivative.

  17. Detection of Campylobacter species using monoclonal antibodies (United States)

    Young, Colin R.; Lee, Alice; Stanker, Larry H.


    A panel of species specific monoclonal antibodies were raised to Campylobacter coli, Campylobacter jejuni and Campylobacter lari. The isotypes, and cross-reactivity profiles of each monoclonal antibody against an extensive panel of micro- organisms, were determined.

  18. Quantum dots-based quantitative and in situ multiple imaging on ki67 and cytokeratin to improve ki67 assessment in breast cancer.

    Directory of Open Access Journals (Sweden)

    Jing Ping Yuan

    Full Text Available As a marker for tumor cell proliferation, Ki67 has important impacts on breast cancer (BC prognosis. Although immunohistochemical staining is the current standard method, variations in analytical practice make it difficult for pathologists to manually measure Ki67 index. This study was to develop a fluorescent spectrum-based quantitative analysis of Ki67 expression by quantum-dots (QDs multiple imaging technique.A QDs-based in situ multiple fluorescent imaging method was developed, which stained nuclear Ki67 as red signal and cytoplasmic cytokeratin (CK as green signal. Both Ki67 and CK signals were automatically separated and quantified by professional spectrum analysis software. This technique was applied to tissue microarrays from 240 BC patients. Both Ki67 and CK values, and Ki67/CK ratio were obtained for each patient, and their prognostic value on 5-year disease free survival was assessed.This method simultaneously stains nuclear Ki67 and cytoplasmic CK with clear signal contrast, making it easy for signal separation and quantification. The total fluorescent signal intensities of both Ki67 sum and CK sum were obtained, and Ki67/CK ratio calculated. Ki67 sum and Ki67/CK ratio were each attributed into two grades by X-tile software based on the best P value principle. Multivariate analysis showed Ki67 grade (P = 0.047 and Ki67/CK grade (P = 0.004 were independent prognostic factors. Furthermore, area under curve (AUC of ROC analysis for Ki67/CK grade (AUC: 0.683, 95%CI: 0.613-0.752 was higher than Ki67 grade (AUC: 0.665, 95%CI: 0.596-0.734 and HER-2 gene (AUC: 0.586, 95%CI: 0.510-0.661, but lower than N stage (AUC: 0.760, 95%CI: 0.696-0.823 and histological grade (AUC: 0.756, 95%CI: 0.692-0.820 on predicting the risk for recurrence.A QDs-based quantitative and in situ multiple imaging on Ki67 and CK was developed to improve Ki67 assessment in BC, and Ki67/CK grade had better performance than Ki67 grade in predicting prognosis.

  19. P53, p63 and Ki-67 assessment in HPV-induced cervical neoplasia. (United States)

    Vasilescu, Florina; Ceauşu, M; Tănase, Cristina; Stănculescu, Ruxandra; Vlădescu, Teodora; Ceauşu, Zenaida


    Carcinoma of the uterine cervix is the most frequent malignancy in women, with an incidence of approximately 456.000 cases per year, leading to 200.000 deaths per year. Twenty-six archived formalin-fixed paraffin-embedded samples of squamous cell carcinoma, selected from 30 Papanicolaou-positive smears, have been analyzed using standard HE stain and the IHC indirect tristadial ABC peroxidase method for four antibodies: p53, p63, Ki-67, HPV. Statistical analysis has been done using the Student t-test, one-group two tails, "paired two samples for mean" variant. Two thirds of the cases were medium and poor differentiated carcinomas. The expression pattern of the proliferation and prognostic factors was biologically correlated with the histopathological type and HPV-infection. Two statistically significant correlations were found between p63 and Ki-67 and between p63 and p53 (pKi-67, showed better correlation with cancer progression than p53. This observation could be used in clinical practice with the purpose of identifying those patients requiring more aggressive treatment.

  20. Expression of Ki-67, Bcl-2 and Bax in the First Trimester Abortion Materials

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    Ender DÜZCAN


    Full Text Available Objective: The aim of this study was to investigate possible similar or different mechanisms in recurrent and spontaneous abortion by evaluating immunohistochemical correlation between proliferation marker Ki-67, and apoptosis markers Bcl-2 and Bax in the fetal trophoblasts and maternal deciduas from abortion material.Material and Method: Eighty samples of curettage materials from 65 abortion patients histopathologically diagnosed “decidua showing Arias-Stella reaction and chorionic villi” or only “decidua showing Arias-Stella reaction” were included in the study. Hematoxylin&Eosin stained sections from all cases were re-evaluated and further stained immunohistochemically using antibodies against Ki-67, Bcl-2 and Bax.Results: Proliferation rate evaluated by Ki-67 expression both in the cytotrophoblastic cells and decidua was found to be significantly lower in spontaneous and recurrent abortions compared to evacuation abortion. The extent of Bcl-2 expression in syncytiotrophoblastic cells covering villous stroma was also decreased in spontaneous abortion. There were no significant differences between spontaneous and recurrent abortions in terms of Bcl-2 expression in syncytiotrophoblasts and Ki-67 proliferation index in cytotrophoblastic cells or decidua. Bax staining showed minimal decidual expression in a few spontaneous and recurrent abortions.Conclusion: We concluded that proliferation rate was decreased in fetal villous cytotrophoblasts and maternal deciduas in spontaneous and recurrent abortions. We also proposed that loss of Bcl-2 expression in syncytiotrophoblasts may cause abortion in a subset of cases. However, the data from spontaneous and recurrent abortions did not not support the presence of different mechanisms in both groups.

  1. Antigen retrieval using pH 3.5 glycine-HCl buffer or urea solution for immunohistochemical localization of Ki-67. (United States)

    Shi, S R; Chaiwun, B; Young, L; Imam, A; Cote, R J; Taylor, C R


    A new antibody (MIB-1) has been described, permitting the demonstration of Ki-67 proliferation antigen in paraffin sections. However, satisfactory results were obtained only after subjecting tissue sections to microwave based antigen retrieval in citrate buffer solution. Other buffer solutions produce equivalent or better results and also permit use of the original Ki-67 antibody, which hitherto has been considered ineffective for paraffin sections.

  2. A monoclonal antibody against leptin. (United States)

    Mahmoudian, Jafar; Jeddi-Tehrani, Mahmood; Bayat, Ali Ahmad; Mahmoudi, Ahmad Reza; Vojgani, Yasaman; Tavangar, Banafsheh; Hadavi, Reza; Zarei, Saeed


    Leptin is an important protein that regulates energy storage and homeostasis in humans and animals. Leptin deficiency results in various abnormalities such as diabetes, obesity, and infertility. Producing a high affinity monoclonal antibody against human leptin provides an important tool to monitor and trace leptin function in different biological fluids. In this study, recombinant human leptin was conjugated to KLH and injected into mice. After immunization, mouse myeloma SP2/0 cells were fused with murine splenocytes followed by selection of antibody-producing hybridoma cells. After screening of different hybridoma colonies by ELISA, a high affinity antibody was selected and purified by affinity chromatography. The affinity constant of the antibody was measured by ELISA. Western blot, immunocytochemistry, and flow cytometry experiments were used to characterize the antibody. The anti-leptin antibody had a high affinity (around 1.13 × 10(-9) M) for its antigen. The saturation of the antibody with leptin (20 moles leptin per 1 mole antibody) in Western blot analysis proved that the antibody had specific binding to its antigen. Immunocytochemistry and flow cytometry on JEG-3 (human placental choriocarcinoma cell) cells revealed that the anti-leptin antibody recognized intracellular leptin. In conclusion, we report here the production and characterization of a murine anti-leptin antibody with high affinity for human leptin.

  3. Ki-67 proliferation index and clinicopathological patterns in colorectal carcinomas

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    Bhagya Lakshmi A


    Full Text Available Background : Tumour, Node, Metastasis (TNM staging system provides useful prognostic information in patients with colorectal carcinoma (CRC. An improved prognostication and patient survival may be achieved by employing immunohistochemistry studies with proliferation markers like Ki-67. Materials and methods: We prospectively studied 51 patients with CRC and evaluated the clinicopathological patterns of CRC and the relationship of with the clinicopathological variables Results: Their mean age was 48 (range 17-75 years; majority (64.7% were males. Rectum was the most common subsite affected (45.1%. Histopathologically most of the tumours (86.3% were usual type adenocarcinomas and were of grade 1 morphology (51%. The Ki-67 proliferation index (PI ranged from 8.4% to 84.4%. The mean PI was greater in patients aged less than or equal to 50 years than in those aged above 50 years, in males than females, in rectal cancers than colonic cancers. It was greater in mucinous carcinomas than usual type adenocarcinomas, in grade 3 tumours than lower grade tumours (grades 1 and 2 and in T4 than T3 and T2 tumours. There was a significant positive correlation between the PI values and grade of the tumour. Conclusion: We concluded that Ki-67 proliferation marker may be useful as an additional tool to assess the tumour aggressiveness with respect to certain clinicopathological parameters in colorectal carcinomas.

  4. Ki67 staining index and neuroendocrine differentiation aggravate adverse prognostic parameters in prostate cancer and are characterized by negligible inter-observer variability. (United States)

    Gunia, Sven; Albrecht, Knut; Koch, Stefan; Herrmann, Thomas; Ecke, Thorsten; Loy, Volker; Linke, Jörg; Siegsmund, Michael; May, Matthias


    This study aims to clarify whether neuroendocrine differentiation (NED) and/or proliferation activity assessed by means of Ki67 staining index (Ki67SI) might aggravate other established adverse prognostic parameters commonly used for predicting outcome in surgically treated prostate cancer, and to assess inter-observer variability in assigning NED and Ki67 SI. A total of 528 patients surgically treated due to prostate cancer were evaluated in this study. Relevant data were retrospectively obtained by chart review. Immunostaining with antibodies directed against Chromogranin A and Ki67 was performed on archived surgical material, and was evaluated by two independent histopathologists blinded to the specimens. Surveying a median postsurgical follow-up of 46.4 months, postsurgical serum PSA-levels were regularly documented for identifying biochemical progress. Multivariate analysis was performed by means of the Cox regression hazards regression method to evaluate possible aggravations of established adverse prognostic parameters (nodal status, tumour stage, pretherapeutic PSA-level, and Gleason score) by NED and/or Ki67SI. Ki67 SI and NED were shown to significantly aggravate these established adverse prognostic parameters, and were found to be characterized by negligible inter-observer variability. Ki67 SI and NED should be advocated to be rendered by the histopathologist because both parameters can be immunohistochemically determined without much additional expense in time and cost involved. This concept is rewarded by an additional gain of prognostic accuracy in evaluating individual risk profile after surgery.

  5. Advances in monoclonal antibody application in myocarditis

    Institute of Scientific and Technical Information of China (English)

    Li-na HAN; Shuang HE; Yu-tang WANG; Li-ming YANG; Si-yu LIU; Ting ZHANG


    Monoclonal antibodies have become a part of daily preparation technologies in many laboratories.Attempts have been made to apply monoclonal antibodies to open a new train of thought for clinical treatments of autoimmune diseases,inflammatory diseases,cancer,and other immune-associated diseases.This paper is a prospective review to anticipate that monoclonal antibody application in the treatment of myocarditis,an inflammatory disease of the heart,could be a novel approach in the future.In order to better understand the current state of the art in monoclonal antibody techniques and advance applications in myocarditis,we,through a significant amount of literature research both domestic and abroad,developed a systematic elaboration of monoclonal antibodies,pathogenesis of myocarditis,and application of monoclonal antibodies in myocarditis.This paper presents review of the literature of some therapeutic aspects of monoclonal antibodies in myocarditis and dilated cardiomyopathy to demonstrate the advance of monoclonal antibody application in myocarditis and a strong anticipation that monoclonal antibody application may supply an effective therapeutic approach to relieve the severity of myocarditis in the future.Under conventional therapy,myocarditis is typically associated with congestive heart failure as a progressive outcome,indicating the need for alternative therapeutic strategies to improve long-term results.Reviewing some therapeutic aspects of monoclonal antibodies in myocarditis,we recently found that monoclonal antibodies with high purity and strong specificity can accurately act on target and achieve definite progress in the treatment of viral myocarditis in rat model and may meet the need above.However,several issues remain.The technology on howto make a higher homologous and weak immunogenic humanized or human source antibody and the treatment mechanism of monoclonal antibodies may provide solutions for these open issues.If we are to further stimulate

  6. Clinical impact of ki-67 labeling index in non-small cell lung cancer

    DEFF Research Database (Denmark)

    Jakobsen, Jan Nyrop; Sørensen, Jens Benn


    The ki-67 index is a marker of proliferation in malignant tumors. Studies from the period 2000 to 2012 on the prognostic and predictive value of ki-67 labeling index (LI) in non-small cell cancer (NSCLC) are reviewed. Twenty-eight studies reported on the prognostic value of ki-67 index with various...

  7. In vivo assessment of the antiproliferative properties of interferon-alpha during immunotherapy: Ki-67 (MIB-1) in patients with metastatic renal cell carcinoma (mRCC)

    DEFF Research Database (Denmark)

    Donskov, Frede; Marcussen, Niels; Hokland, M.


    -alpha-based immunotherapy in 34 patients and repeated after 5 weeks in 25 patients. Tumour proliferation was assessed by use of the anti-Ki-67 antibody MIB-1 and evaluated in multiple, random systematic sampled fields of vision. Ki-67 labelling index (LI) at baseline was median 13.6% (range 1.2-85.0) and median 10.......016). Baseline or change in Ki-67 LI did not correlate to survival. These data suggest that IFN-alpha in vivo has only modest effect on tumour proliferation in patients with mRCC. Tumour Ki-67 (MIB-1) reactivity after 1 month of immunotherapy appears to be a significant predictor of patient survival....

  8. Comparative immunoexpression of ICAM-1, TGF-β1 and ki-67 in periapical and residual cysts (United States)

    Armada, Luciana; dos Santos, Teresa-Cristina; Pires, Fabio-Ramoa


    Background This study compared the immunohistochemical expression of ki-67, transforming growth factor beta 1 (TGF-β1) and intercellular adhesion molecule-1 (ICAM-1) in inflammatory periapical cysts and residual cysts. Material and Methods The study sample was composed by 25 periapical cysts and 25 residual cysts and immunohistochemical reactions were carried out using antibodies directed against ICAM-1, TGF-β1 and ki-67. Clinical, radiological, gross, histological and immunohistochemical data were tabulated for descriptive and comparative analysis using the SPSS software and differences were considered statistically significant when p<0.05%. Results There were no differences between the expression of ICAM-1 (p=0.239) and TGF-β1 (p=0.258) when comparing both groups. Ki-67 labeling index was higher in residual cysts compared to periapical cysts (p=0.017). Conclusions Results from the present study suggest that some specific inflammatory stimuli on residual cysts would modulate their mechanisms of etiopathogenesis, growing and repair. Key words:Periapical cyst, radicular cyst, residual cyst, transforming growth factor beta 1 (TGF-β1), intercellular adhesion molecule 1 (ICAM-1), ki-67. PMID:27918735

  9. Pharmacokinetics interactions of monoclonal antibodies. (United States)

    Ferri, Nicola; Bellosta, Stefano; Baldessin, Ludovico; Boccia, Donatella; Racagni, Giorgi; Corsini, Alberto


    The clearance of therapeutic monoclonal antibodies (mAbs) typically does not involve cytochrome P450 (CYP450)-mediated metabolism or interaction with cell membrane transporters, therefore the pharmacokinetics interactions of mAbs and small molecule drugs are limited. However, a drug may affect the clearance of mAbs through the modulation of immune response (e.g., methotrexate reduces the clearance of infliximab, adalimumab, and golimumab, possibly due to methotrexate's inhibitory effect on the formation of antibodies against the mAbs). In addition, mAbs that are cytokine modulators may modify the metabolism of drugs through their effects on P450 enzymes expression. For example, cytokine modulators such as tocilizumab (anti-IL-6 receptor antibody) may reverse the "inhibitory" effect of IL-6 on CYP substrates, resulting in a "normalization" of CYP activities. Finally, a drug may alter the clearance of mAbs by either increasing or reducing the levels of expression of targets of mAbs on the cell surface. For instance, statins and fibrates induce PCSK9 expression and therefore increase cellular uptake and clearance of alirocumab and evolocumab, anti-PCSK9 antibodies. In the present review, we will provide an overview on the pharmacokinetics properties of mAbs as related to the most relevant examples of mAbs-small molecule drug interaction.

  10. Ki-67 is required for maintenance of cancer stem cells but not cell proliferation (United States)

    Cidado, Justin; Wong, Hong Yuen; Rosen, D. Marc; Cimino-Mathews, Ashley; Garay, Joseph P.; Fessler, Abigail G.; Rasheed, Zeshaan A.; Hicks, Jessica; Cochran, Rory L.; Croessmann, Sarah; Zabransky, Daniel J.; Mohseni, Morassa; Beaver, Julia A.; Chu, David; Cravero, Karen; Christenson, Eric S.; Medford, Arielle; Mattox, Austin; De Marzo, Angelo M.; Argani, Pedram; Chawla, Ajay; Hurley, Paula J.; Lauring, Josh; Park, Ben Ho


    Ki-67 expression is correlated with cell proliferation and is a prognostic marker for various cancers; however, its function is unknown. Here we demonstrate that genetic disruption of Ki-67 in human epithelial breast and colon cancer cells depletes the cancer stem cell niche. Ki-67 null cells had a proliferative disadvantage compared to wildtype controls in colony formation assays and displayed increased sensitivity to various chemotherapies. Ki-67 null cancer cells showed decreased and delayed tumor formation in xenograft assays, which was associated with a reduction in cancer stem cell markers. Immunohistochemical analyses of human breast cancers revealed that Ki-67 expression is maintained at equivalent or greater levels in metastatic sites of disease compared to matched primary tumors, suggesting that maintenance of Ki-67 expression is associated with metastatic/clonogenic potential. These results elucidate Ki-67's role in maintaining the cancer stem cell niche, which has potential diagnostic and therapeutic implications for human malignancies. PMID:26823390

  11. Production and Screening of Monoclonal Peptide Antibodies. (United States)

    Trier, Nicole Hartwig; Mortensen, Anne; Schiolborg, Annette; Friis, Tina


    Hybridoma technology is a remarkable and indispensable tool for generating high-quality monoclonal antibodies. Hybridoma-derived monoclonal antibodies not only serve as powerful research and diagnostic reagents, but have also emerged as the most rapidly expanding class of therapeutic biologicals. In this chapter, an overview of hybridoma technology and the laboratory procedures used routinely for hybridoma production and antibody screening are presented, including characterization of peptide antibodies.

  12. Expressions of Ki-67 and vascular endothelial growth factor in lichen planus%扁平苔藓中Ki-67和VEGF的表达

    Institute of Scientific and Technical Information of China (English)

    王娟; 白莉; 鲍海平; 郑爱义; 武晓华; 赵一锦; 邓起; 米希婷


    Objective To investigate the expressions of Ki-67 and vascular endothelial growth factor (VEGF) in lichen planus and explore their effects on the pathogenesis of lichen planus. Methods The expressions of Ki-67 and VEGF in 20 lichen planus and 20 normal skin specimens were detected by immunohistochemical method. Results The expressions Ki-67 and VEGF were significantly higher in lichen planus than those in normal skins (P < 0.01). There was a positive correlation between the expression of Ki-67 and VEGF in lichen planus (r = 0.74, P < 0.01). Conclusions The up-regulated expressions of Ki-67 and VEGF may play an important role in the development of lichen planus.%目的:研究扁平苔藓皮损中Ki-67和血管内皮生长因子(VEGF)的表达情况,探讨它们在扁平苔藓发病中的作用.方法:采用免疫组化法检测20例扁平苔藓皮损中Ki-67和VEGF的表达情况,20例正常皮肤作为对照.结果:扁平苔藓组中Ki-67和VEGF的表达水平明显高于对照组,差别具有统计学意义(P < 0.01);扁平苔藓皮损中Ki-67和VEGF的表达呈正相关(r = 0.74,P < 0.01).结论:Ki-67、VEGF的异常高表达可能参与了扁平苔藓的发病.

  13. Monoclonal antibodies in chronic lymphocytic leukemia. (United States)

    Ferrajoli, Alessandra; Faderl, Stefan; Keating, Michael J


    Multiple options are now available for the treatment of chronic lymphocytic leukemia. Over the last 10 years, monoclonal antibodies have become an integral part of the management of this disease. Alemtuzumab has received approval for use in patients with fludarabine-refractory chronic lymphocytic leukemia. Rituximab has been investigated extensively in chronic lymphocytic leukemia both as a single agent and in combination with chemotherapy and other monoclonal antibodies. Epratuzumab and lumiliximab are newer monoclonal antibodies in the early phase of clinical development. This article will review the monoclonal antibodies more commonly used to treat chronic lymphocytic leukemia, the results obtained with monoclonal antibodies as single agents and in combination with chemotherapy, and other biological agents and newer compounds undergoing clinical trials.

  14. Evaluation of cervical cone biopsies for coexpression of p16INK4a and Ki-67 in epithelial cells. (United States)

    Reuschenbach, Miriam; Seiz, Mirjam; von Knebel Doeberitz, Christina; Vinokurova, Svetlana; Duwe, Alexander; Ridder, Ruediger; Sartor, Heike; Kommoss, Friedrich; Schmidt, Dietmar; von Knebel Doeberitz, Magnus


    Diffuse overexpression of p16(INK4a) in basal and parabasal cells of cervical epithelium is a hallmark of human papillomavirus-mediated transformation. Focal p16(INK4a) expression is occasionally observed in nondysplastic epithelium. In normal cells, expression of p16(INK4a) triggers cell cycle arrest. However, cells undergoing transformation in intraepithelial lesions actively proliferate. To prove that the different expression patterns of p16(INK4a) , i.e., focal versus diffuse, reflect biologically different entities, we hypothesized that p16(INK4a) -positive cells in epithelia displaying focal p16(INK4a) expression pattern do not coexpress proliferation-associated Ki-67 protein, while p16(INK4a) -positive cells in lesions with diffuse p16(INK4a) expression may do. A total of 138 cervical cone biopsies were stained for the expression of p16(INK4a) and Ki-67 using a primary antibody cocktail. All metaplastic lesions (n = 21) displayed focal staining for p16(INK4a) , and in all of these lesions p16(INK4a) -positive cells were found to be negative for Ki-67 expression. Diffuse expression of p16(INK4a) was observed in 12/21 (57.1%) cervical intraepithelial neoplasia (CIN) 1 lesions, all of them simultaneously showed Ki-67 immunoreactivity in a large proportion of p16(INK4a) -positive cells. Seventeen of 23 (73.9%) CIN2 lesions and all 27 (100%) CIN3/carcinoma in situ (CIS) as well as all 46 (100%) carcinoma cases displayed diffuse and combined expression of p16(INK4a) and Ki-67. Coexpression of Ki-67 and p16(INK4a) in the same cell is entirely restricted to cervical lesions displaying diffuse p16(INK4a) expression, whereas in lesions with focal p16(INK4a) expression, p16(INK4a) -expressing cells are negative for Ki-67. Thus, diffuse expression of p16(INK4a) reflects lesions with proliferation-competent cells, while p16(INK4a) -expressing cells associated with focal expression patterns are cell cycle arrested.

  15. Expression and significance of Ki-67 in psoriatic lesions%Ki-67在寻常型银屑病中的表达及其意义

    Institute of Scientific and Technical Information of China (English)

    林新瑜; 何飞; 董丹丹; 李科


    Objective To investigate the effect of Ki-67 0n the pathogenesis of psoriasis vulgaris lesions and to evaluate its possible significance. Methods The expression and distribution of Ki-67 were detected by immunohistochemistry in the lesions of 48 cases with psoriasis vulgaris and in the epidermis of 20 normal controls. The mRNA expression of Ki-67 was detected by RT-PCR methods. Results The expressions of Ki-67 in psoriatic epidermis were significantly higher than those in normal epidermis ; they were located in all epidermal layers except horny layer. There were no or only weak positive expressions in the basal layer of normal controls. Ki-67 mRNA were markedly increased in psoriasis vulgaris. Conclusion The over expressions of Ki-67 in psoriatic epidermis suggest that it might be related to the hyperproliferation and ahnormal differentiation of psoriatic keratinocytes. Ki-67 may be a novel target in preventing the process of psoriasis vulgaris.%目的 探讨Ki-67在寻常型银屑病发病中的作用.方法 采用免疫组织化学LDP法检测了48例寻常型银屑病皮损区及20例正常人表皮中Ki-67的表达及分布特点,采用逆转录聚合酶链反应技术(RT-PCR方法)捡测Ki-67的mRNA表达水平.结果 寻常型银屑病皮损区Ki-67表达均较正常表皮中的表达明显增强,分布于除角质层以外的表皮各层,而正常表皮表达阴性或仅在基底层有弱阳性表达.RT-PCR检测结果表明寻常型银屑病组Ki-67的表达水平亦显著高于正常表皮.结论 Ki-67在寻常型银屑病皮损中过度表达,提示其与银屑病的表皮细胞过度增殖、分化异常有关.

  16. Ki-67 expression in axillary lymph node metastases in breast cancer is prognostically significant. (United States)

    Tawfik, Kareem; Kimler, Bruce F; Davis, Marilyn K; Fan, Fang; Tawfik, Ossama


    Several studies have documented the prognostic significance of cell proliferation in breast cancer and its positive relationship with tumor grade, size, mitotic activity, hormonal and Her-2 status, and tumor progression. The Ki-67 antigen provides an accurate measure of the growth fraction of a tumor. Ki-67 expression in 103 primary breast carcinomas and their corresponding axillary lymph node metastases was correlated with age, tumor grade, size, estrogen receptor (ER), progesterone receptor (PgR), p53, epidermal growth factor receptor (EGFR), Bcl-2, Her-2 status, and patients' overall survival. Median Ki-67 expression in primary and metastatic tumors was 20% and 15%, respectively. Although there was no difference in overall survival (P = .65, log-rank test) between primary tumors with less than or at least 10% Ki-67 expression, there was significantly better overall survival when Ki-67 expression in lymph nodes was less than 10% (P = .040). For patients whose primary tumors exhibited Ki-67 expression less than 10%, most of their metastatic lesions had a similar low Ki-67; these patients had a favorable outcome. A small subgroup was noted to have a nodal Ki-67 of 10% or more and worse survival (P = .047). For patients whose primary tumors had a Ki-67 of 10% or more, most of their metastatic lesions had similar high Ki-67 values; however, a group of 12 patients had lymph node Ki-67 less than 10% and had a better overall survival (P = .092). Our results showed that measurement of Ki-67 in lymph node is superior to its evaluation in primary tumors. Identification of subgroups of patients in whom Ki-67 expression in lymph nodes differs from expression in primary tumor may assist in the selection of therapeutic options.

  17. Associação entre a expressão das proteínas p53 e Ki-67 e os achados clínico-patológicos em pacientes com carcinoma invasor do colo uterino Association between p53 and Ki-67 expression and clinicopathologic features in patients with carcinoma of the cervix

    Directory of Open Access Journals (Sweden)

    Agnaldo L. Silva-Filho


    analyzed by histopathology (hematoxylin and eosin and immunohistochemically evaluated using monoclonal antibodies for p53 and Ki-67. Data were analyzed statistically by the chi2 test to evaluate eventual differences between the groups. RESULTS: the age of the patients ranged from 27 to 73 years (48.7±10.4 years. Clinical stage (FIGO was IB1 in 27 cases (75% and IB2 in 9 cases (25%. A positive tumoral expression of the p53 protein was found in half of the cases. In relation to the Ki-67 expression, a high cell proliferation index was shown in 73.3% of the cases. There was no association between tumoral p53 and Ki-67 expression with age (p=0.091 and 0.900, clinical stage (p=0.054 and 0.667, histological classification (p=0.674 and 0.674, grade of differentiation (p=0.070 and 0.282, presence of lymphatic vascular space invasion (p=0.248 and 0.667, parametrial involvement (p=0.729 and 0.763 and pelvic lymph node metastasis (p=0.729 and 0.636, respectively. CONCLUSIONS: tumoral expression of p53 and Ki-67 was not associated with the clinicopathological features in patients with stage IB carcinoma of the cervix.

  18. Effect of 1,25-dihydroxyvitamin D3 influence on expressions of Ki67 and mTOR in Thy-1 nephritis model of rat%1,25(OH)2D3对Thy-1肾炎大鼠Ki67、mTOR表达的影响

    Institute of Scientific and Technical Information of China (English)

    杨锐; 索洁; 李建锋; 王文政; 赵瑾; 陶林; 杨晓萍


    Objective To study the expressions of Ki67 and mTOR in Thy-1 nephritis model of rat who were given 1,25-dihydroxyvitamin D3[1,25(OH)2D3] and to explore its mechanism. Methods Healthy male SD rats (n=90) were random⁃ly divided into three groups: control group, model group, 1,25(OH)2D3 treatment group (n=30 in each group). Model group and 1,25(OH)2D3 treatment group were intravenously injected with anti-Thy1 monoclonal antibody once via tail vein while the control group were administrated with same volume of normal saline through the same route. 1,25(OH)2D3 were adminis⁃trated at 0.5μg per day intra-gastrically for consecutive 21 days in 1,25(OH)2D3 treatment group while equal volume of pea⁃nut oil were given in control group and model group. Six rats were randomly selected from each group and sacrificed at the 1st , 3rd , 7th , 14th and 21st after drug intervention. Twenty four hour urine sample were collected in each rat just before it was culled to detect 24-hour urinary protein excretion. Renal tissue samples were harvested and stained with hematoxylin&eo⁃sin (H&E) and PAS to determine the renal pathological variation and the expressions of mTOR and Ki67 were assessed by immunohistochemistry. Results Urine protein begin to be detected at the first day after model was established, peaked at the 3rd days then started dropping until the 14th day when urine sample turned to normal. Urine protein levels were lower in 1, 25(OH)2D3 treatment group at the 1st,3rd,7th day after model establishment than those in model group(P<0.05). Compared with model group, the pathological damage of renal tissue in 1,25(OH)2D3 treatment group were alleviated at the 3rd and 7th day after model establishment (P < 0.05). Expressions of Ki67 and mTOR in 1, 25(OH)2D3 treatment group were reduced compared with those in model group (P<0.05). Twenty four hour urinary protein and expressions of Ki67 and mTOR as well as renal pathological damage were all positively correlated with each

  19. Computerized image analysis of the Ki-67 proliferation index in mantle cell lymphoma

    DEFF Research Database (Denmark)

    Blaker, Yngvild N; Brodtkorb, Marianne; Maddison, John;


    practice. The aim of this study was to investigate the use of computerized image analysis software for scoring of Ki-67 in MCL. METHODS AND RESULTS: We developed an automated method for determining the Ki-67 index by computerized image analysis and tested it using a cohort of 62 MCL patients. The data were...... compared to Ki-67 scores obtained by semiquantitative estimation and image-based manual counting. When using the Ki-67 index as a continuous parameter, both image-based manual counting and computerized image analysis were related inversely to survival (P = 0.020 and P = 0.025, respectively). Ki-67 index...... obtained by semiquantitative estimation was not associated significantly with survival (P = 0.093). The results were validated in a second patient cohort with similar results. CONCLUSION: Computerized image analysis of the Ki-67 index in MCL is an attractive alternative to semiquantitative estimation...

  20. Late stage cathepsin C, CXCL13 and Ki-67 overexpression correlate with regional neuropathology in a BSE transgenic murine model. (United States)

    Vidal, E; Tortosa, R; Marco, P; Fondevila, D; Rabanal, R M; Torres, J M; Pumarola, M


    A DNA microarray-based gene expression analysis study was performed with bovine spongiform encephalopathy (BSE) transgenic mice. Several genes were found to be overexpressed including the lysosomal enzyme cathepsin C, the chemokine CXCL13 and a number of genes related to cellular proliferation. The brains from terminal stage, BSE inoculated, 'bovinized', transgenic mice were subjected to immunohistochemistry with antibodies against these two proteins and Ki-67, a cell proliferation marker, to assess the biological relevance of the gene expression changes. Differential expression of cathepsin C and CXCL13 proteins and increased expression of Ki-67 was observed. These changes were localized to areas of deposition of PrP(res) and spongiform change and to areas showing an astroglial and microglial response. These findings suggest that these proteins are involved in the mechanisms leading to the establishment of transmissible spongiform encephalopathy.

  1. Polyclonal and monoclonal antibodies in clinic. (United States)

    Wootla, Bharath; Denic, Aleksandar; Rodriguez, Moses


    Immunoglobulins (Ig) or antibodies are heavy plasma proteins, with sugar chains added to amino-acid residues by N-linked glycosylation and occasionally by O-linked glycosylation. The versatility of antibodies is demonstrated by the various functions that they mediate such as neutralization, agglutination, fixation with activation of complement and activation of effector cells. Naturally occurring antibodies protect the organism against harmful pathogens, viruses and infections. In addition, almost any organic chemical induces antibody production of antibodies that would bind specifically to the chemical. These antibodies are often produced from multiple B cell clones and referred to as polyclonal antibodies. In recent years, scientists have exploited the highly evolved machinery of the immune system to produce structurally and functionally complex molecules such as antibodies from a single B clone, heralding the era of monoclonal antibodies. Most of the antibodies currently in the clinic, target components of the immune system, are not curative and seek to alleviate symptoms rather than cure disease. Our group used a novel strategy to identify reparative human monoclonal antibodies distinct from conventional antibodies. In this chapter, we discuss the therapeutic relevance of both polyclonal and monoclonal antibodies in clinic.

  2. Relation between qualitative and quantitative 3-dimensional ultrasound and ki-67 expression in breast cancer. (United States)

    Wang, Xiao-Yan; Zhang, Bing; He, Yan; Ning, Bing; Nong, Mei-Fen; Wei, Hai-Ming; Huang, Xiang-Hong


    To investigate the relation between quantitative blood flow parameters on 3-dimensional (3D) color histogram, 3D ultrasound characteristics and Ki-67 expression in breast cancer. Three-dimensional ultrasound characteristics and histological classifications of 76 breast tumors in 75 confirmed cases were analyzed. Relations of tumor volume (V), vascularization index (VI), flow index (FI) and vascularization-flow index (VFI) on 3D color histogram to Ki-67 expression were studied by statistical methods. VI and VFI measurements of tumors in positive Ki-67 expression group were obviously increased compared with the negative expression group (P0.05). Cases showing positive expression of Ki-67 were more likely to have lymph node metastases (P0.05). Qualitative and quantitative 3D ultrasound characteristics correlated with positive expression of Ki-67 in breast cancer. Quantitative analysis with 3D color histogram more accurately evaluates blood supply of breast tumors, providing references for predicting biological behaviors and prognosis of breast cancer.

  3. Role of Ki-67 in acromegalic patients with hyperprolactinemia: retrospective analysis in 61 Chinese Patients. (United States)

    Huan, Cheng; Cui, Guihua; Lu, Chao; Qu, Xin; Han, Tao


    To evaluate the specific characteristics in acromegalic patients with hyperprolactinemia by analyzing the differences between patients with different Ki-67 values. Between 2002 and 2010, a set of data on 61 patients undergoing transsphenoidal surgery was available at the Department of Neurosurgery, Provincial Hospital Affiliated to Shandong University. Patients were divided into Ki-67 >3% group and 3% group had a higher rate of invasiveness (P=0.048), higher incidences of diabetes mellitus (P=0.036), coarse facial features (P=0.048), large hands and feet (P=0.003), higher GH levels (P<0.05), higher diabetes insipidus rate (P<0.001), and more frequent recurrence (P=0.011) than Ki-67 <3% group. Patients with higher Ki-67 value harbored longer time interval, more aggressive tumors, more acromegaly manifestations, higher GH level, and higher recurrence than patients with lower Ki-67 value.

  4. Expression of Survivin, CDK4, Ki-67 and Clinical Significance in Pediatric Acute Leukemia

    Institute of Scientific and Technical Information of China (English)

    ZHANG Liuqing; LIU Jing; LIN Hanhua; HU Qun; LIU Aiguo; HU Ying


    The expression of Survivin, CDK4 and Ki-67 and the clinical significance in pediatric acute leukemia (AL) were investigated. The expression of Survivin, CDK4 and Ki-67 proteins was detected by using immunohistochemical assay in 37 children with AL and 10 children with normal bone marrow as controls. The positive expression rate of Survivin, CDK4 and Ki-67 was 45.9 %,56.8 %, and 40.5 % respectively in 37 AL children, which was significantly higher than in control group accordingly (P<0.05). The expression of Survivin was positively correlated with CDK4(P=0.007) and Ki-67 (P=0.008). In conclusion, all Survivin, CDK4 and Ki-67 proteins are over-expressed in pediatric AL and involved in the modulation of apoptosis and proliferation in pediatric AL.

  5. Expression of the p16 and Ki67 in Cervical Squamous Intraepithelial Lesions and Cancer. (United States)

    Kanthiya, Kanjana; Khunnarong, Jakkapan; Tangjitgamol, Siriwan; Puripat, Napaporn; Tanvanich, Sujitra


    To evaluate the expression of p16 and Ki67 in cervical intraepithelial neoplasia (CIN) and cancer. We performed a immunohistochemical study of p16 and Ki67 in 243 cervical tissues 53 nondysplastic lesions, 106 CIN1, 61 CIN2/3 and 23 squamous cell carcinomas. The expression of p16 and Ki67 was interpreted independently by 2 researchers and the sensitivity and specificity to detect clinically significant lesions (≥ CIN2) were determined. The overall agreement results of positive or negative immunostaining of intrainter observer variability were 0.659 for p16 and 0.808 for Ki67. p16 expression was demonstrated in 91.3% of invasive carcinomas, 78.7% of CIN2/3, 10.4% of CIN1 and 9.4% of nondysplasic lesions. The corresponding Ki67 expression was: 100% of all invasive carcinomas, 75.4% of CIN2/3, 22.6% of CIN1, and 11.3% with nondysplasia. The expression was significantly different between CIN2/3 vs CIN1 for both p16 and Ki67 (pvalues cancer vs CIN2/3 for Ki67 (pvalue 0.008). The differences were not significant between CIN1 vs nondysplasia (pvalues 1.000 for p16 and 0.130 of Ki67), and cancer vs CIN2/3 for p16 (p value 0.219). The sensitivity and specificity to detect > CIN2 were 84.5% and 90.5% by p16 and 82.1% and 88.6% by Ki67. The rates for 16 and Ki67 expression were directly associated with the severity of cervical lesions. Significant differences in these markers expression may be useful in cases with equivocal histologic features among cervical intraepithelial lesions, but not between CIN1 and nondysplastic lesions. The two markers had high sensitivity and specificity in determining >CIN2.

  6. Variable Ki67 proliferative index in 65 cases of nodular fasciitis, compared with fibrosarcoma and fibromatosis. (United States)

    Lin, Xu-Yong; Wang, Liang; Zhang, Yong; Dai, Shun-Dong; Wang, En-Hua


    Nodular fasciitis is the most common pseudosarcomatous lesion of soft tissue. Ki67 was considered as a useful marker for distinguishing some benign and malignant lesions. To study the usefulness of Ki67 in diagnosis of nodular fasciitis, the expression of Ki67 was examined by using immunostaining in 65 nodular fasciitis specimens, 15 desmoid fibromatosis specimens and 20 fibrosarcoma specimens. The results showed that there was a variable Ki67 index in all 65 cases of nodular fasciitis, and the mean labeling index was 23.71±15.01%. In majority (70.77%) of all cases,the index was ranged from 10% to 50%, in 6.15% (4/65) of cases the higher Ki67 index (over 50%) could be seen. The Ki67 proliferative index was closely related to duration of lesion, but not to age distribution, lesion size, sites of lesions and gender. Moreover, the mean proliferative index in desmoid fibromatosis and fibrosarcoma was 3.20±1.26% and 26.15±3.30% respectively. The mean Ki67 index of nodular fasciitis was not significantly lower than fibrosarcoma, but higher than desmoid fibromatosis. The variable and high Ki67 index in nodular fasciitis may pose a diagnostic challenge. We should not misdiagnose nodular fasciitis as a sarcoma because of its high Ki67 index. The recurrence of nodular fasciitis is rare; and the utility of Ki67 immunostaining may be not suitable for recurrence assessment in nodular fasciitis. The virtual slide(s) for this article can be found here:

  7. Monoclonal antibodies reactive with hairy cell leukemia

    NARCIS (Netherlands)

    Visser, L; Shaw, A; Slupsky, J; Vos, H; Poppema, S

    Monoclonal antibodies reactive with hairy cell leukemia were developed to aid in the diagnosis of this subtype of B cell chronic lymphocytic leukemia and to gain better insight into the origin of hairy cells. Three antibodies were found to be of value in the diagnosis of hairy cell leukemia.

  8. Monoclonal Antibody Therapy for Advanced Neuroblastoma (United States)

    NCI is sponsoring two clinical trials of a monoclonal antibody called ch14.18, in combination with other drugs, to see if the antibody may be helpful for children or young adults (up to age 21) with relapsed or refractory neuroblastoma.

  9. Monoclonal antibodies reactive with hairy cell leukemia

    NARCIS (Netherlands)

    Visser, L; Shaw, A; Slupsky, J; Vos, H; Poppema, S


    Monoclonal antibodies reactive with hairy cell leukemia were developed to aid in the diagnosis of this subtype of B cell chronic lymphocytic leukemia and to gain better insight into the origin of hairy cells. Three antibodies were found to be of value in the diagnosis of hairy cell leukemia. Antibod

  10. Production of Monoclonal Antibody against Human Nestin. (United States)

    Hadavi, Reza; Zarnani, Amir Hassan; Ahmadvand, Negah; Mahmoudi, Ahmad Reza; Bayat, Ali Ahmad; Mahmoudian, Jafar; Sadeghi, Mohammad-Reza; Soltanghoraee, Haleh; Akhondi, Mohammad Mehdi; Tarahomi, Majid; Jeddi-Tehrani, Mahmood; Rabbani, Hodjattallah


    We have employed a peptide-based antibody generation protocol for producing antibody against human nestin. Using a 12-mer synthetic peptide from repetitive region of human nestin protein devoid of any N- or O-glyco-sylation sequences, we generated a mouse monoclonal antibody capable of recognizing human, mouse, bovine, and rat nestin. A wide variety of nestin proteins ranging from 140-250 kDa was detected by this antibody. This antibody is highly specific and functional in applications such as ELISA, flow cytometry, immunocytochemistry, and Western blot assays.

  11. P53 and Ki-67 as prognostic markers in triple-negative breast cancer patients (United States)

    Pan, Yunbao; Yuan, Yufen; Liu, Guoshi; Wei, Yongchang


    Triple-negative breast cancer (TNBC) is an aggressive subgroup of breast cancer lack of effective target therapy. This study was to investigate the prognostic role of p53 and Ki-67 in 156 cases of TNBC patients. Logistic regression analysis was used to examine the association between clinical parameters and recurrence. Univariate and multivariate analyses were used to examine the association between clinical characteristics and disease-free survival (DFS) or overall survival (OS). Survival analyses using the Kaplan-Meier method were performed to examine the association between p53/Ki-67 and DFS and OS. Our data showed that p53 was positive in 71.3% and the Ki-67 high index was in 82.8% of TNBC. Elevated p53 and Ki-67 were associated with histological grade. The tumor size, lymph node involvement, and p53 expression are associated with risk of recurrence. Tumor size, lymph node involvement, family history, Ki-67 and p53 are independent variables associated with either DFS or OS. TNBC patients with positive p53 or Ki-67 high index or family history of cancer have a significant association with worse prognosis. This study suggests that p53, Ki-67 and family history are useful prognostic markers in TNBC. PMID:28235003

  12. A study of Ki-67 expression and its clinicopathological determinants in nondysplastic oral leukoplakia (United States)

    Mondal, Krishnendu; Mandal, Rupali; Sarkar, Badal Chandra


    Context: Oral cancer is the third most prevalent malignancy in India. Leukoplakia is its most common precursor lesion. Aims: This study aimed at evaluation of the Ki-67 expression and thereby detection of the dysplastic potential in histopathologically nondysplastic oral leukoplakia (OL). Secondarily, another purpose was to correlate various clinicopathological factors with the labeling indices (LIs) of Ki-67 in those cases as well. Settings and Design: In total, 97 OL cases were examined. Relevant clinical and demographic information was retrieved from the pro forma, prefilled by the patients themselves during their first visit. Subjects and Methods: Ki-67 immunohistochemical staining was performed on paraffin-embedded tissue samples. Its LIs were calculated and correlated with different clinicopathological parameters using statistical software SPSS version 16.0. Results: 58.8% (57 cases) lesions exhibited a Ki-67 positivity of ≤5%, and 25.8% (25 cases) lesions exhibited it in the range of 6%–25%. Only 15 (15.4%) patches were stained positively between 26% and 60%. Patients’ age beyond 50 years, nonhomogeneous leukoplakia, and tobacco addiction were the significant risk factors for high Ki-67 scores (P < 0.05). Conclusions: Ki-67 is an essential immunohistochemical marker for epithelial dysplasia in OL, especially when the conventional histopathology fails to appreciate the same. In this purpose, Ki-67 labeling on a routine basis delivers the most convenient results for patients aged above 50 years, and/or addicted to tobacco products, and/or suffering from nonhomogeneous patches. PMID:27994417

  13. Correlations of β-catenin, Ki67 and Her-2/neu with gastric cancer

    Institute of Scientific and Technical Information of China (English)

    Hong-Wen Wu; Cheng-Yong Qin; Ji-Lai Huang; Xian-Yi Kong; Wen-Ji Wang; Wen-Kun Bai


    Objective: To study the clinical pathologic characteristics of β-catenin, Ki67 and Her-2/neu in gastric cancer and the correlation of β-catenin and Ki67 to the protein expression and gene conditions of Her-2/Neu. Methods: The protein expression of β-catenin, Ki67 and Her-2/Neu was detected by immunohistochemistry in 101 cases of gastric cancer and the gene conditions of Her-2/Neu by fluorescence in situ hybridization (FISH). Results: The protein expression ofβ-catenin, Ki67 and Her-2/Neu had close relationship with the clinical pathologic characteristics of gastric cancer. The β-catenin and Ki67 had obvious correlation to the differentiation, infiltration and lymphatic metastasis of the gastric cancer (P<0.05). The Ki67 had close relationship with the tumor-node-metastasis staging staging of gastric cancer (P<0.05). Her-2/Neu had close relationship with the differentiation and tumor-node-metastasis staging of gastric cancer (P<0.05) but had no relationship with the infiltration and lymphatic metastasis of the gastric cancer (P<0.05). The protein expression of Ki67 had significantly positive correlation to the protein expression and gene amplification conditions of Her-2/Neu (r=0.567, P<0.05 for protein; r=0.304, P<0.05 for gene). Conclusions: Combined detection of β-catenin, Ki67 and Her-2/Neu can be used as a reliable method to help the observation of biological behavior, diagnosis and prognosis of gastric cancer, and Ki67 can be used to serve the preliminary screening of Her-2/Neu gene state.

  14. Immunohistochemical Ki-67/KL1 double stains increase accuracy of Ki-67 indices in breast cancer and simplify automated image analysis

    DEFF Research Database (Denmark)

    Nielsen, Patricia S; Bentzer, Nina K; Jensen, Vibeke


    observers and automated image analysis. RESULTS: Indices were predominantly higher for single stains than double stains (P≤0.002), yet the difference between observers was statistically significant (PPearson correlation coefficient for manual and automated indices ranged from 0.......69 to 0.85 (Pcorrelating automated indices with tumor characteristics, for example, tumor size (P... stains, Ki-67 should be quantified on double stains to reach a higher accuracy. Automated indices correlated well with manual estimates and tumor characteristics, and they are thus possibly valuable tools in future exploration of Ki-67 in breast cancer....

  15. Ki-67 Expression in Human Tumors Measured by Flow Cytometry (United States)


    proliferation. One of the first proliferation antigens to be studied was the transferrin receptor (TfR). Proliferating normal and tumor cells require iron and...obtained by incubation in NP-40. When the antibody was used to stain frozen sections of human tonsil , the chromosomes were stained. The antibody was...proliferation. When applied to frozen sections of human tonsil , the antibody appeared to be reactive with a mitotic spindle-associated protein. It bound

  16. Demonstration of the proliferation marker Ki-67 in renal biopsies: correlation to clinical findings. (United States)

    Nabokov, A; Waldherr, R; Ritz, E


    Assessment of cell proliferation in renal biopsy samples is a potentially promising analytical tool to evaluate disease activity. So far no information is available on the correlation between proliferative activity in different anatomic compartments of the kidney and clinical symptoms. To elucidate this issue, we examined renal biopsy specimens from 20 patients with systemic vasculitis (15 Wegener's granulomatosis, five microscopic polyangiitis), 20 patients with immunoglobulin (Ig) A nephropathy (IgAN), 13 patients with minimal-change disease (MCD), 11 patients with tubulointerstitial nephritis, and five patients with diabetes mellitus. The streptavidin-biotin-peroxidase complex technique was applied to autoclave-pretreated, formalin-fixed, paraffin-embedded tissue sections to label different cell types with the antibody MIB1 directed against the Ki-67 antigen. Proliferation index (PI) was estimated as the number of positively stained nuclei per glomerular cross-section or per square millimeter section area. The interstitial cells were discriminated by additional staining of Ki-67-processed samples with specific immune markers. In patients with vasculitis, PI was considerably elevated in the extracapillary glomerular compartment (0.86), in proximal tubules (6.24), and in the interstitium (8.62). High proliferative activity was also noted in interstitium (3.98) and proximal tubules (1.35) of patients with IgAN. Of particular interest was the increased interstitial proliferative activity (15.0) in diabetic patients. Resident renal cells, but not infiltrating cells, seemed to constitute the majority of the proliferating cell population in the interstitium. In systemic vasculitis, clinical disease activity was significantly correlated to endocapillary (r(s) = 0.58), extracapillary (r(s) = 0.67), proximal tubular (r(s) = 0.67), and interstitial PI (r(s) = 0.61). By multiple linear regression analysis, proximal tubular PI was correlated to the presence of hematuria

  17. Proliferating cell nuclear antigen and Ki-67 immunohistochemistry of oligodendrogliomas with special reference to prognosis

    DEFF Research Database (Denmark)

    HEEGAARD, S.; Sommer, Helle Mølgaard; BROHOLM, H.;


    Background. The biologic behavior of oligodendrogliomas is somewhat unpredictable. A supplementary prognostic factor is, therefore, desirable. Methods. Thirty-two pure supratentorial oligodendrogliomas were investigated using proliferating cell nuclear antigen (PCNA) and Ki-67 immunohistochemical...

  18. Immunohistochemical patterns of follicular dendritic cell meshwork and Ki-67 in small B-cell lymphomas

    Institute of Scientific and Technical Information of China (English)



    Objective To identify the immunohistochemical patterns of follicular dendritic cell(FDC)meshwork and Ki-67labeling index in small B-cell lymphomas(SBLs) and their significance in differential diagnosis.Methods

  19. Expression of PPARγ, p27 and Ki67 in Cer vical Cancer and its Clinical Signiifcance

    Institute of Scientific and Technical Information of China (English)

    LI Peng-li; TAN Hai-zhi


    Objective: To investigate the expression of peroxisome proliferation-activated receptor γ(PPARγ), p27 and Ki67 in cervical cancer and its clinical signiifcance. Methods:The expression of PPARγ, p27 and Ki67 in the tissues of 42 patients with cervical cancer, 28 with cervical intraepithelial neoplasia (CIN) and 12 with normal cervix was detected using immunohistochemistry. Results:The positive rate of PPARγprotein in cervical cancer tissue (76.2%) was signiifcantly higher than in CIN (53.6%) and normal cervical tissue (8.3%) (P0.05). Conclusion:Abnormal expression of PPARγ, p27 and Ki67 may play important roles in occurrence and progression of cervical cancer, and hence, joint detection of PPARγ, p27 and Ki67 can be used to diagnose early CIN and cervical cancer.

  20. Significance of IMP3, nucleophosmin, and Ki-67 expression in papillary thyroid carcinoma. (United States)

    Yorukoglu, Aygun; Yalcin, Nagihan; Avci, Arzu; Cakalagaoglu, Fulya; Yaylali, Guzin; Akin, Fulya; Haciyanli, Mehmet; Ozden, Akin


    The purpose of our study was to investigate the diagnostic value of expression of IMP3, nucleophosmin, and correlation of these markers with Ki-67 proliferation index in papillary thyroid carcinoma and benign neoplasms of thyroid gland. The aim was also to investigate whether there is a difference between papillary and micropapillary carcinomas with regard to clinicopathologic parameters beside IMP3, nucleophosmin, and Ki-67 proliferation index. It was concluded that IMP3 and nucleophosmin cannot be a routine diagnostic marker for discrimination of papillary carcinomas and benign lesions. IMP3 positive staining was quite scarce in IMP3 positive papillary carcinomas although specifity of IMP3 is 100%. A statistically significant correlation was not detected between nucleophosmin, IMP-3, and Ki-67 proliferation index. A statistically significant correlation was found between tumor size, lymphovascular embolism, and Ki-67 proliferation index. There was also significant correlation between tumor size and lymphovascular embolism.

  1. Ki-67 proliferation index in patients with placenta previa percreta in the third trimester. (United States)

    Hilali, Nese; Kocarslan, Sezen; Vural, Mehmet; Incebiyik, Adnan; Camuzcuoglu, Aysun; Camuzcuoglu, Hakan


    The purpose of this study was to investigate proliferative capacity of placenta previa percreta in the third trimester via evaluating Ki-67 proliferating index. The paraffin blocks of placental tissues, which were obtained from the patients who underwent hysterectomy for placenta previa percreta (n = 12, gestational age > 28 weeks), from legal abortions (n = 12, gestational age  38 weeks), between January 2011 and April 2013, were included into the study. The paraffin blocks of the patients were stained with Ki-67 (proliferating cell marker) immunohistochemically, and Ki-67 proliferation index levels were calculated. Ki-67 proliferation index levels were higher in patients with legal abortions than patients with placenta percreta or noncomplicated cesarean delivery group. However, any statistically significant difference was not detected between the percreta and noncomplicated groups (p > 0.05). The tissue samples of the patients with placenta previa percreta exhibited low proliferative capacity similar to the samples of normal placentation group.

  2. [Clinical value of p16/Ki-67 immunocytochemical dual staining in cervical cancer screening]. (United States)

    Wang, H R; Liao, G D; Chen, W; Qiao, Y L; Jiang, Y


    Objective: to investigate the clinical value of p16/Ki-67 immunocytochemical dual staining (abbreviated as p16/Ki-67 dual staining) in cervical intraepithelial neoplasia (CIN) and cervical cancer screening. Methods: From July to November 2015, a total of 980 women attending cervical cancer screening and receiving high-risk human papillomavirus (HR-HPV) test and thinprep cytologic test (TCT) were included in the study. p16/Ki-67 immunocytochemical dual staining was performed on residual cytologic specimens and compared with histopathology results. Results: The expression risks of p16/Ki-67 in HPV16/18 group and another HR-HPV group were higher than HPV negative group, with an odds ratio of 10.64 (95%CI: 5.66~20.02) and 5.40 (95%CI: 3.62~8.04), respectively. The positive rate of p16/Ki-67 increased with the grade of TCT and histologic diagnosis, and was higher in both CIN2 and CIN3 groups than normal group (Pp16/Ki-67 to detect CIN2+ and CIN3+ lesions was 89.3% and 94.1%, respectively, and the specificity was 69.3% and 66.8%, respectively. The sensitivity of TCT to detect CIN2+ and CIN3+ lesions was 60.7% and 64.7%, respectively, and the specificity was 49.3% and 49.1%, respectively. Conclusions: Compared with TCT, p16/Ki-67 dual staining has higher sensitivity and specificity. It can identify high-grade cervical lesions and guide the classification of CIN. p16/Ki-67 dual staining in conjunction with HPV test may be considered as an efficient method for cervical cancer screening.

  3. Microvessel density and Ki-67 labeling index in esthesioneuroblastoma: is there a prognostic role? (United States)

    Singh, Lavleen; Ranjan, Richa; Madan, Renu; Arava, Sudheer K; Deepak, Rakesh K; Singh, Manoj Kumar


    Esthesioneuroblastoma (ENB) is a malignant neuroectodermal tumor. Hyams grading has an established role in its prognostication. The importance of microvessel density (MVD) and Ki-67 labeling index (Ki-67 LI) is well studied in various tumors, but the same remains understated in ENB. The aims of the study were to estimate proliferation index and MVD in ENB and to correlate them with Hyams grade. Twenty-six ENB cases diagnosed over a period of 5 years were included. Hyams grade, MVD, and Ki-67 LI were evaluated for each of them. The cases were categorized as low (Hyams grades 1 and 2) and high (Hyams grades 3 and 4) grades. Microvessel density and Ki-67 LI were correlated with grade. The treatment response was analyzed in different grades. The commonest histologic grade was 4 (42%). The mean Ki-67 LI was 2%, 8.2%, 30.8%, and 40.5% and mean MVD was 81.67/mm(2), 37/mm(2), 24/mm(2), and 25.2/mm(2) in grades 1, 2, 3, and 4, respectively. A statistically significant correlation of grade with Ki-67 LI (P < .001) and MVD (P < .007) was noted. Hyams grade in ENB correlates well with treatment response. Ki-67 LI is an important prognostic factor in ENB. We propose a cutoff of 25% for Ki-67 LI to differentiate low- vs high-grade ENB, but larger studies are needed for validation. Contrary to epithelial tumors, there is a decrease in MVD with increasing grade in ENB.

  4. Ki-67 overexpression in WHO grade II gliomas is associated with poor postoperative seizure control. (United States)

    Yuan, Yang; Xiang, Wang; Yanhui, Liu; Ruofei, Liang; Shuang, Liu; Yingjun, Fan; Qiao, Zhou; Yanwu, Yang; Qing, Mao


    Seizures are the most common initial symptom in patients with low-grade gliomas, and approximately 30% of these patients still suffer from epilepsy after gross-total resection of the tumour. We examined the relationship between the overexpression of ki-67 in WHO grade II gliomas and seizure control. A series of 93 histologically confirmed WHO grade II glioma tissues were analysed through immunohistochemical staining for ki-67 expression. Follow-up visits regarding seizure control were scheduled at 12 months. The Engel classification was used to categorise patients' seizure status. Of the 93 patients analysed, 65 (66.3%) patients initially presented with seizures. A total of 36 patients were diagnosed with WHO grade II oligodendrogliomas, 29 patients had oligoastrocytomas and 28 patients had astrocytomas. Ki-67 was over-expressed in 15 patients. One year after surgery poor seizure control was observed in 11 of these patients. In contrast, low ki-67 expression (seizure control was observed in 36 patients (difference between ki-67 over- and low expression groups P = 0.002). Logistic regression analysis revealed that patients with gross-total resection achieved better seizure control while ki-67 overexpression and age below 38 years were poor seizure control factors explained of the variance of seizure outcome (OR: 0.382, 4.354 and 1.822, respectively). In WHO grade II gliomas, Ki-67 is a molecular marker which predicts poor seizure control of glioma patients after the resection of the tumour. Gross-total resection, ki-67 overexpression and age below 38 years significantly affect seizure prognosis. Copyright © 2013 British Epilepsy Association. Published by Elsevier Ltd. All rights reserved.


    Directory of Open Access Journals (Sweden)

    V. P. Avdoshin


    Full Text Available The prognostic value of the expression Ki-67, ММР-9 and collagen IV were estimated in urotelial bladder tumors. The biopsy and surgery samples from 43 patients (27 males and 16 females aged 42 to 87 years (mean age 66 ± 1.5 years who received combination treatment for urotelial tumors. It has been found that Ki-67, ММР-9 and collagen IV are important prognostic markers of urotelial invasive bladder carcinoma.

  6. Monoclonal antibody technologies and rapid detection assays (United States)

    Novel methodologies and screening strategies will be outlined on the use of hybridoma technology for the selection of antigen specific monoclonal antibodies. The development of immunoassays used for diagnostic detection of prions and bacterial toxins will be discussed and examples provided demonstr...

  7. Cyclooxygenase-2 and Ki67 Expression in Oral Leukoplakia: a Clinicopathological Study

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    Alper Sinanoglu


    Full Text Available Objectives: Oral leukoplakia is a precancerous lesion of the oral mucosa. The upregulation of Ki67 and cyclooxygenase-2 has been reported in both dysplastic and non-dysplastic tissues. The aim of this clinicopathological study was to investigate the prognostic value of Ki67 and cyclooxygenase-2 expression for oral leukoplakia. Material and Methods: A total of 50 samples were investigated and the study group consisted of 30 oral leukoplakia samples. Samples of 10 intact oral mucosa and 10 squamous cell carcinoma were included as negative and positive control groups, respectively. Epithelial dysplasia was defined as oral intraepithelial neoplasia (OIN and classified into subgroups 1 - 3. Tissue samples were assessed immunohistochemically for Ki67 and cyclooxygenase-2 expression. Clinicopathological correlations of oral leukoplakia patients were also investigated. Results: All OIN 3 patients were non-smokers (P < 0.05, and homogeneous oral leukoplakia lesions also presented OIN. Both cyclooxygenase-2 and Ki67 expression increased with the severity of lesions, which defined different subgroups (P < 0.05, except there was no significant difference between the hyperkeratosis and OIN groups for Ki67 expression. Conclusions: Cyclooxygenase-2 and Ki67 expression may have a prognostic value for the malignant transformation of oral leukoplakia.

  8. Evaluation of potential prognostic value of Bmi-1 gene product and selected markers of proliferation (Ki-67 and apoptosis (p53 in the neuroblastoma group of tumors

    Directory of Open Access Journals (Sweden)

    Katarzyna Taran


    Full Text Available Introduction: Cancer in children is a very important issue in pediatrics. The least satisfactory treatment outcome occurs among patients with clinically advanced neuroblastomas. Despite much research, the biology of this tumor still remains unclear, and new prognostic factors are sought. The Bmi-1 gene product is a currently highly investigated protein which belongs to the Polycomb group (PcG and has been identified as a regulator of primary neural crest cells. It is believed that Bmi‑1 and N-myc act together and are both involved in the pathogenesis of neuroblastoma. The aim of the study was to assess the potential prognostic value of Bmi-1 protein and its relations with mechanisms of proliferation and apoptosis in the neuroblastoma group of tumors.Material/Methods: 29 formalin-fixed and paraffin-embedded neuroblastoma tissue sections were examined using mouse monoclonal antibodies anti-Bmi-1, anti-p53 and anti-Ki-67 according to the manufacturer’s instructions.Results: There were found statistically significant correlations between Bmi-1 expression and tumor histology and age of patients.Conclusions: Bmi-1 seems to be a promising marker in the neuroblastoma group of tumors whose expression correlates with widely accepted prognostic parameters. The pattern of BMI-1 expression may indicate that the examined protein is also involved in maturation processes in tumor tissue.

  9. Monoclonal antibodies to Treponema Pallidum.

    NARCIS (Netherlands)

    H.J.M. van de Donk; J.D.A. van Embden; M.F. van Olderen; A.D.M.E. Osterhaus (Albert); J.C. de Jong (Jan)


    textabstractThree successive fusions of mouse myeloma cells and spleen lymphocytes of a mouse immunized with Treponema Pallidum resulted in one hybridoma producing anti T. pallidum antibodies for each fusion. The mice were immunized with live pallidum cells respectively 1, 3 and 5 months before fusi

  10. In vivo assessment of the antiproliferative properties of interferon-alpha during immunotherapy: Ki-67 (MIB-1) in patients with metastatic renal cell carcinoma (mRCC)

    DEFF Research Database (Denmark)

    Donskov, Frede; Marcussen, Niels; Hokland, M.


    The aim of the present study was to investigate the in vivo antiproliferative effect of interferon alpha (IFN-alpha) in patients with metastatic renal cell carcinoma (mRCC). Core needle biopsies of metastatic and/or the primary kidney cancer were obtained before interleukin-2 (IL-2)- and IFN......-alpha-based immunotherapy in 34 patients and repeated after 5 weeks in 25 patients. Tumour proliferation was assessed by use of the anti-Ki-67 antibody MIB-1 and evaluated in multiple, random systematic sampled fields of vision. Ki-67 labelling index (LI) at baseline was median 13.6% (range 1.2-85.0) and median 10.......016). Baseline or change in Ki-67 LI did not correlate to survival. These data suggest that IFN-alpha in vivo has only modest effect on tumour proliferation in patients with mRCC. Tumour Ki-67 (MIB-1) reactivity after 1 month of immunotherapy appears to be a significant predictor of patient survival....

  11. Immunohistochemical characterization of molecular classification of breast carcinoma and its relation with Ki-67

    Directory of Open Access Journals (Sweden)

    Shabnam Karangadan


    Full Text Available Background: Breast carcinoma is the leading cause of cancer deaths in women. Molecular classification of breast carcinoma along with Ki-67 index is considered a better predictive factor for prognosis and treatment than routine histopathology. Aims: To classify breast carcinoma into the four molecular subtypes defined by immunohistochemical expression of triple markers: Luminal A (estrogen receptor/progesterone receptor-positive [ER/PR+] and human epidermal growth factor receptor 2 HER2/neu, luminal B (ER/PR + and HER2/neu+, triple negative (ER/PR − and HER2/neu−, and HER2 positive (ER/PR−, HER2/neu+, and to correlate the expression of ER, PR, HER2/neu, and classification with Ki-67. Materials and Methods: The present study includes sixty breast carcinoma cases studied over a 3-year period. The expression patterns of ER, PR, HER2/neu, and Ki-67 were studied. Clinical features, pathologic features such as size, grade, and lymph node status, and correlation with Ki-67 of the four subtypes were compared. Results: Out of sixty cases, most common molecular subtype was triple negative (40.00% followed by luminal B (23.33%. Most of the tumors showed low proliferative index (low Ki-67; however, triple negative and HER2 positive subtype showed high proliferative index. Most common histological subtype was ductal carcinoma which was mainly triple negative. All medullary carcinoma cases were triple negative. One case of lobular carcinoma and mucinous carcinoma each was HER2 positive and luminal B, respectively. Single case of carcinoma of male breast was luminal B subtype. Conclusion: Correlation of molecular classification with age, histological grade, and Ki-67 was statistically significant (P < 0.05. ER/PR also correlated with histological grade and Ki-67 (P < 0.01. These results emphasize the fact that molecular subtypes correlate with prognosis and aid in targeted therapy.

  12. The importance of tissue handling of surgically removed breast cancer for an accurate assessment of the Ki-67 index (United States)

    Arima, Nobuyuki; Nishimura, Reiki; Osako, Tomofumi; Nishiyama, Yasuyuki; Fujisue, Mamiko; Okumura, Yasuhiro; Nakano, Masahiro; Tashima, Rumiko; Toyozumi, Yasuo


    Aim Insufficient attention for the Ki-67 immunohistochemistry has been given to the importance of tissue handling for surgical breast cancer specimens. We sought to investigate the effect of fixation status on the Ki-67. Methods We examined the effect of fixative, time to and duration of fixation using surgical specimens, and finally, compared the paired Ki-67 index in the tumour between core needle and surgical specimen. Results The Ki-67 was significantly higher when 10% neutral buffered formalin was used (p=0.0276). Insufficient fixation caused a drastic reduction in the Ki-67 index (p=0.0177), but not significant in oestrogen receptor (ER) and human epidermal growth factor receptor 2 (HER2). Sixteen hours delayed time to fixation also caused a reduction of the Ki-67 (p=0.0284), but not significant in ER. Prolonged fixation significantly led to a gradual reduction in the Ki-67 in a time-dependent manner, but not in both ER and HER2. Finally, cutting the tumour before fixation improved fixation status and consequently caused an increased level of the Ki-67 index (p=0.0181), which resulted in a strong correlation of the Ki-67 between core needle and surgical specimen (r=0.8595). Conclusions Tissue handling of surgical specimen is critical for assessing the Ki-67 compared with ER and HER2. We should pay more attention to tissue fixation status for the standard assessment of the Ki-67 index. PMID:26420767

  13. Human Monoclonal Antibodies as a Countermeasure Against Botulinum Toxins (United States)


    REPORT Human monoclonal antibodies as a countermeasure against Botulinum toxins 14. ABSTRACT 16. SECURITY CLASSIFICATION OF: In this report, we...Prescribed by ANSI Std. Z39.18 - 31-Aug-2012 Human monoclonal antibodies as a countermeasure against Botulinum toxins Report Title ABSTRACT In this report...DTRA Final Report: Human monoclonal antibodies as a countermeasure against Botulinum toxins   Page 1 of 22 DTRA Final Report: Human monoclonal

  14. Ki67, chemotherapy response, and prognosis in breast cancer patients receiving neoadjuvant treatment

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    Fasching Peter A


    Full Text Available Abstract Background The pathological complete response (pCR after neoadjuvant chemotherapy is a surrogate marker for a favorable prognosis in breast cancer patients. Factors capable of predicting a pCR, such as the proliferation marker Ki67, may therefore help improve our understanding of the drug response and its effect on the prognosis. This study investigated the predictive and prognostic value of Ki67 in patients with invasive breast cancer receiving neoadjuvant treatment for breast cancer. Methods Ki67 was stained routinely from core biopsies in 552 patients directly after the fixation and embedding process. HER2/neu, estrogen and progesterone receptors, and grading were also assessed before treatment. These data were used to construct univariate and multivariate models for predicting pCR and prognosis. The tumors were also classified by molecular phenotype to identify subgroups in which predicting pCR and prognosis with Ki67 might be feasible. Results Using a cut-off value of > 13% positively stained cancer cells, Ki67 was found to be an independent predictor for pCR (OR 3.5; 95% CI, 1.4, 10.1 and for overall survival (HR 8.1; 95% CI, 3.3 to 20.4 and distant disease-free survival (HR 3.2; 95% CI, 1.8 to 5.9. The mean Ki67 value was 50.6 ± 23.4% in patients with pCR. Patients without a pCR had an average of 26.7 ± 22.9% positively stained cancer cells. Conclusions Ki67 has predictive and prognostic value and is a feasible marker for clinical practice. It independently improved the prediction of treatment response and prognosis in a group of breast cancer patients receiving neoadjuvant treatment. As mean Ki67 values in patients with a pCR were very high, cut-off values in a high range above which the prognosis may be better than in patients with lower Ki67 values may be hypothesized. Larger studies will be needed in order to investigate these findings further.

  15. Monoclonal Antibodies to Plant Growth Regulators (United States)

    Eberle, Joachim; Arnscheidt, Angelika; Klix, Dieter; Weiler, Elmar W.


    Four high affinity monoclonal antibodies, which recognize two plant growth regulators from the cytokinin group, namely trans-zeatin riboside and dihydrozeatin riboside and their derivatives are reported. Six hybridomas were produced from three independent fusions of Balb/c spleen cells with P3-NS1-Ag 4-1 (abbreviated NS1) or X63-Ag 8.653 (X63) myeloma cells. The mice had been hyperimmunized with zeatin riboside-bovine serum albumin conjugate or dihydrozeatin riboside-bovine serum albumin conjugate for 3 months. The hybridomas secrete antibodies of the IgG 1 or IgG 2b subclass and allow the detection of femtomole amounts of the free cytokinins, their ribosides, and ribotides in plant extracts. The use of these monoclonals in radio- and enzyme-linked immunosorbent assay is also discussed. PMID:16664848

  16. Recent developments in monoclonal antibody radiolabeling techniques

    Energy Technology Data Exchange (ETDEWEB)

    Srivastava, S.C.; Mease, R.C.


    Monoclonal antibodies (MAbs) have shown the potential to serve as selective carriers of radionuclides to specific in vivo antigens. Accordingly, there has been an intense surge of research activity in an effort to develop and evaluate MAb-based radiopharmaceuticals for tumor imaging (radioimmunoscintigraphy) and therapy (radioimmunotherapy), as well as for diagnosing nonmalignant diseases. A number of problems have recently been identified, related to the MAbs themselves and to radiolabeling techniques, that comprise both the selectivity and the specificity of the in vivo distribution of radiolabeled MAbs. This paper will address some of these issues and primarily discuss recent developments in the techniques for radiolabeling monoclonal antibodies that may help resolve problems related to the poor in vivo stability of the radiolabel and may thus produce improved biodistribution. Even though many issues are identical with therapeutic radionuclides, the discussion will focus mainly on radioimmunoscintigraphic labels. 78 refs., 6 tabs.

  17. An immunohistochemical analysis of sex-steroid receptors, tumor suppressor gene p53 and Ki-67 in the normal and neoplastic uterine cervix squamous epithelium

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    Nikolaou Marinos


    Full Text Available Introduction. Malignant transformation of sex-steroid dependent tissues is associated with the loss of expression of sex steroid receptors as well as of the tumor suppression gene p53. The aim of this study is to evaluate the expression of sex-steroid receptors, p53 and Ki-67 in specimens from pre-malignant and malignant cervical epithelial lesions throughout the menstrual cycle. Material and Methods. Immunohistochemical staining was performed on formalin fixed, paraffin embedded tissue sections of normal squamous cervical epithelium, cervical intraepithelial neoplasia and invasive squamous cervical carcinoma, specimens utilizing antibodies against estrogen receptors, progesterone receptors, p53 protein and Ki-67 antigen. Results. In the samples taken from the normal cervical tissue, basal cells were usually estrogen receptor-positive, progesterone receptornegative, p53-negative and Ki-67-negative throughout the menstrual cycle. In contrast, para-basal cells were estrogen receptorpositive and progesterone receptor-negative in the follicular phase, but estrogen receptor-negative and progesterone receptor -positive and Ki-67 positive in the luteal phase. In cervical precancerous and cancer tissue samples (cervical intraepithelial neoplasia and squamous cervical carcinoma, the expression of estrogen receptors decreased. 31.15% of cervical intraepithelial neoplasia and 11.5% of squamous cervical carcinoma were positive for estrogen receptors. However, the expression of progesterone receptors increased. 29.5% of cervical intraepithelial neoplasia and 49.2% of squamous cervical carcinoma were positive for progesterone receptors. Positive staining for p53 was observed in 15 (24.59% cases of cervical intraepithelial neoplasia and in 39 (64% of squamous cervical carcinoma. The expression Ki-67 index in squamous cervical carcinoma cases (47.60% was significantly higher than of cervical intraepithelial neoplasia cases (30.2% (p=0.041. Conclusion. The

  18. Assay for the specificity of monoclonal antibodies in crossed immunoelectrophoresis

    DEFF Research Database (Denmark)

    Skjødt, K; Schou, C; Koch, C


    A method is described based on crossed immunoelectrophoresis of a complex antigen mixture in agarose gel followed by incubation of the gel with the monoclonal antibody. The bound monoclonal antibody is detected by the use of a secondary enzyme-labelled antibody. Using this technique we have been...... I molecules. In other experiments using the same technique we demonstrated the reaction of a monoclonal antibody specific for chicken Ig light chains. Udgivelsesdato: 1984-Aug-3...

  19. Typical and atypical bronchopulmonary carcinoid tumors: a clinicopathologic and KI-67-labeling study. (United States)

    Costes, V; Marty-Ané, C; Picot, M C; Serre, I; Pujol, J L; Mary, H; Baldet, P


    Expression of a proliferating antigen by KI-67 immunohistochemistry was analyzed with a SAMBA 2005 computer-assisted image processor (Traitement de l'Information for des Techniques Nouvelles, Grenoble, France) in 47 surgically resected bronchopulmonary carcinoids embedded in paraffin. The clinicopathologic characteristics and KI-67 labeling, expressed in percentage of stained nuclear surface relative to the total nuclear surface, of 31 typical carcinoids and 16 atypical carcinoids were compared and assessed with respect to patient survival. The proliferation status was significantly higher in histologically atypical than in typical carcinoids. Moreover, using a 4% cutoff, we observed a significant difference for the 4-year overall survival rate. Semiquantitative analysis of the proliferation index by KI-67 immunostaining seemed to be an effective means of identifying high risk subsets among patients with histologically atypical carcinoids and for whom adjuvant chemotherapy could be proposed.

  20. Therapeutic monoclonal antibody for Sporotrichosis

    Directory of Open Access Journals (Sweden)

    Sandro eAlmeida


    Full Text Available Sporotrichosis is a chronic subcutaneous mycosis that affects either humans or animals and occurs worldwide. This subcutaneous mycosis had been attributed to a single etiological agent, Sporothrix schenckii. S. schenckii exhibits a considerable genetic variability, where recently, was suggesting that this taxon consists of a complex of species. Sporotrichosis is caused by traumatic inoculation of the fungus, which is a ubiquitous environmental saprophyte that can be isolated from soil and plant debris. The infection is limited to the cutaneous forms but, recently, occurrences of more severe clinical forms of this mycosis were described, especially among immunocompromized individuals. The immunological mechanisms involved in prevention and control of sporotrichosis are still not very well understood. Some works suggest that cell-mediated immunity plays an important role in protecting the host against S. schenckii. In contrast, the role of the humoral immune response in protection against this fungus have not been studied in detail. In a previous study, we showed that antigens secreted by S. schenckii induce a specific humoral response in infected animals, mainly against the 70-kDa molecules, indicating a possible participation of specific antibodies to this molecule in infection control. In an other work of the our group, we produced a mAb against a 70-kDa glycoprotein of S. schenckii in order to better understand the effect of passive immunization of mice infected with S. schenckii. Results showed a significant reduction in the number of CFU in organs of mice when the mAb was injected before and during S. schenckii infection. Similar results were observed when T-cell deficient mice were used. Drugs of choice in the treatment of sporothrichosis require long periods and frequently relapses are observed, mainly in immunocompromized patients. The strong protection induced by mAb against a 70-kDa glycoprotein makes it a strong candidate for a

  1. Correlation of apparent diffusion coefficient with Ki-67 proliferation index in grading meningioma. (United States)

    Tang, Yi; Dundamadappa, Sathish K; Thangasamy, Senthur; Flood, Thomas; Moser, Richard; Smith, Thomas; Cauley, Keith; Takhtani, Deepak


    A noninvasive method to predict aggressiveness of high-grade meningiomas would be desirable because it would help anticipate tumor recurrence and improve tumor management and the treatment outcomes. The Ki-67 protein is a marker of tumor proliferation, and apparent diffusion coefficient (ADC) is related to tumor cellularity. Therefore, we sought to determine whether there is a statistically significant correlation between ADC and Ki-67 values in meningiomas and whether ADC values can differentiate various meningioma subtypes. MRI examinations and histopathology of 68 surgically treated meningiomas were retrospectively reviewed. Mean ADC values were derived from diffusion imaging. Correlation coefficients were calculated for mean ADC and Ki-67 proliferation index values using linear regression. An independent unpaired Student t test was used to compare the ADC and Ki-67 proliferation index values from low-grade and more aggressive meningiomas. A statistically significant inverse correlation was found between ADC and Ki-67 proliferation index for low-grade and aggressive meningiomas (r(2) = -0.33, p = 0.0039). ADC values (± SD) of low-grade meningiomas (0.84 ± 0.14 × 10(-3) mm(2)/s) and aggressive (atypical or anaplastic) meningiomas (0.75 ± 0.03 × 10(-3) mm(2)/s) were significantly different (p = 0.0495). Using an ADC cutoff value of 0.70 × 10(-3) mm(2)/s, the sensitivity for diagnosing aggressive meningiomas was 29%, specificity was 94%, positive predictive value was 67%, and negative predictive value was 75%. ADC values correlate inversely with Ki-67 proliferation index and help differentiate low-grade from aggressive meningiomas.

  2. Frequency of Ki-67 (MIB-1 and P53 expressions among patients with prostate cancer

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    Seyed Hamid Madani


    Full Text Available Context: Prostate cancer is the most common malignant tumor in men. Tumor grade is one of the most important prognostic factors of prostate cancer. P53 and Ki-67 expressions have also been considered to be prognostic factors. Aims: This study was performed to investigate the frequency of these proteins expression and compare the obtained results with Gleason′s grading. Settings and Design: In this cross-sectional study, 49 paraffin blocks of prostate cancers were assessed. Tumor grade was determined according to the Gleason′s criteria. Materials and Methods: Ki-67 and P53 expressions were determined by immunohistochemical staining. Statistical Analysis: The obtained results were analyzed and evaluated using Spearman′s statistical test (SPSS version 15. Results: Three out of 49 (6.1% cases were well differentiated, 21 (43% moderately differentiated and 25 (51% were poorly differentiated. P53 was negative in all well-differentiated cases. Ki-67 was negative in 14 cases (28% including all well-differentiated tumors. Among moderately and poorly differentiated tumors Ki-67 was negative in eight (38% and three (12% of cases, respectively. A statistically significant relation was observed between the increased Ki-67 labeling index (LI and increased Gleason′s grade. Conversely, no statistically significant relation was found between P53 expression and increased Gleason′s grade. Conclusions: According to the findings of this study, it seems that Ki-67 can be used as a prognostic factor for prostate cancer. On the other hand, the probable relation between P-53 and prostate cancer prognosis requires further studies.

  3. Expression of Progestrone Receptor and Proliferative Marker ki 67, in Various Grades of Meningioma

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    Nasrin Shayanfar


    Full Text Available Meningiomas are slow-growing neoplasms which recur locally, their morphologic grading is simple but do not always correlate with patient outcome. The aim of present study is to evaluate the status of progesterone receptor (PR and proliferation marker Ki67 in various grades of meningioma in a group of Iranian patients. 78 cases of meningioma were selected from the file of a hospital university. All archival H&E stained sections were reviewed and regraded according to WHO criteria. Immunohistochemical analysis for PR and Ki67 was performed on formalin- fixed, paraffin- embedded samples. PR status considered positive if > 10% of tumor cell's nuclei were strongly immunoreactive, or if > 50% of nuclei were stained with medium intensity. The Ki67 labeling index (LI is defined as the percentage area with strongest immunostaining. PR were positive in 61/63(96.8% of grade I tumors,2/10(20% of grade II,and 0/5(0% of grade III tumors.Ki67 LI was %2.98±2.27 in grade I tumors, %9.30±5.79 in grade II tumors and %34.00±5.47 in grade III tumors. For both markers, differences between grade I,II and III tumors were significant(P<0.001.There was a reverse relationship between mean of Ki67 LI and PR status,with increasing grade of tumor. Evaluation of PR status and Ki67 LI together with conventional histologic evaluation can help in providing more information about the biologic behaviour of meningiomas,especially for those that histological grading is not straightforward.

  4.  Immunohistochemical Expression of ki-67 and p53 in Colorectal Adenomas: A Clinicopathological Study

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    Hussam Hasson Ali


    Full Text Available  Objectives: To evaluate the significance of P53 and Ki-67 expression as immunohistochemical markers in early detection of premalignant changes in different types of colorectal adenomas. Also, to correlate immunohistochemical expression of the two markers with different clinicopathological parameters including; age, and sex of the patient, type, site, size and grade of dysplasia of colorectal adenomas.Methods: Forty-seven polypectomy specimens of colorectal adenomas were retrieved from the archival materials of the Gastrointestinal and Hepatic Diseases Teaching Hospital in Baghdad from 2009 - 2010. Four µm section specimens were stained by immunohistochemical technique with Ki-67 and P53 tumor markers. P-values <0.05 were considered statistically significant.Results: Immunohistochemical expressions of Ki-67 and P53 had a significant correlation with the size and grade of dysplasia in colorectal adenomas. However, there was no significant correlation among the immunohistochemical expression of Ki-67 and P53 with the age and gender of the patient, and the type and site of colorectal adenomas. There was no significant correlation between Ki-67 and P53 expressions in colorectal adenomas. Villous adenomas of colorectum showed a significant correlation with the grade of dysplasia, while there was no significant correlation between size and site of colorectal adenoma with the grade of dysplasia.Conclusion: High grade dysplasia with significant positive immunohistochemical markers of Ki-67 and P53 could be valuable parameters for selecting from the total colorectal adenoma population, those most deserving of close surveillance in follow-up cancer prevention programs. It is closely linked with increasing age particularly in patients with a large size adenoma of villous component in their histology.

  5. Expression of PPARγ, p27 and Ki67 in Cervical Cancer and its Clinical Significance

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    Peng-li LI


    Full Text Available Objective: To investigate the expression of peroxisome proliferation-activated receptor γ (PPARγ, p27 and Ki67 in cervical cancer and its clinical significance. Methods: The expression of PPARγ, p27 and Ki67 in the tissues of 42 patients with cervical cancer, 28 with cervical intraepithelial neoplasia (CIN and 12 with normal cervix was detected using immunohistochemistry. Results:The positive rate of PPARγ protein in cervical cancer tissue (76.2% was significantly higher than in CIN (53.6% and normal cervical tissue (8.3% (P<0.05 orP<0.01, which was also evidently higher in CIN than in normal cervical tissue (P<0.05. The positive rate of p27 protein in cervical cancer tissue (31.0% was significantly lower than in CIN (57.1% and normal cervical tissue (83.3% (P<0.05 or P<0.01, and that in CIN had a markedly lower tendency compared with normal cervical tissue (P<0.05. The positive rate of Ki67 protein in cervical cancer tissue (100.0% was apparently higher than in CIN (85.7% and normal cervical tissue (33.3% (P<0.05 or P<0.01, which was also markedly higher in CIN than in normal cervical tissue (P<0.01. The expression of PPARγ, p27 and Ki67 proteins was not associated with the clinicopathological features of patients, including the age, histological types, pathological grading and clinical staging (P>0.05.Conclusion: Abnormal expression of PPARγ, p27 and Ki67 may play important roles in occurrence and progression of cervical cancer, and hence, joint detection of PPARγ, p27 and Ki67 can be used to diagnose early CIN and cervical cancer.

  6. Predicting Ki67% expression from DCE-MR images of breast tumors using textural kinetic features in tumor habitats (United States)

    Chaudhury, Baishali; Zhou, Mu; Farhidzadeh, Hamidreza; Goldgof, Dmitry B.; Hall, Lawrence O.; Gatenby, Robert A.; Gillies, Robert J.; Weinfurtner, Robert J.; Drukteinis, Jennifer S.


    The use of Ki67% expression, a cell proliferation marker, as a predictive and prognostic factor has been widely studied in the literature. Yet its usefulness is limited due to inconsistent cut off scores for Ki67% expression, subjective differences in its assessment in various studies, and spatial variation in expression, which makes it difficult to reproduce as a reliable independent prognostic factor. Previous studies have shown that there are significant spatial variations in Ki67% expression, which may limit its clinical prognostic utility after core biopsy. These variations are most evident when examining the periphery of the tumor vs. the core. To date, prediction of Ki67% expression from quantitative image analysis of DCE-MRI is very limited. This work presents a novel computer aided diagnosis framework to use textural kinetics to (i) predict the ratio of periphery Ki67% expression to core Ki67% expression, and (ii) predict Ki67% expression from individual tumor habitats. The pilot cohort consists of T1 weighted fat saturated DCE-MR images from 17 patients. Support vector regression with a radial basis function was used for predicting the Ki67% expression and ratios. The initial results show that texture features from individual tumor habitats are more predictive of the Ki67% expression ratio and spatial Ki67% expression than features from the whole tumor. The Ki67% expression ratio could be predicted with a root mean square error (RMSE) of 1.67%. Quantitative image analysis of DCE-MRI using textural kinetic habitats, has the potential to be used as a non-invasive method for predicting Ki67 percentage and ratio, thus more accurately reporting high KI-67 expression for patient prognosis.

  7. Association of tissue inhibitor of metalloproteinases-1 and Ki67 in estrogen receptor positive breast cancer

    DEFF Research Database (Denmark)

    Bjerre, Christina Annette; Knoop, Ann; Bjerre, Karsten;


    Background. The role of tissue inhibitor of metalloproteinases-1 (TIMP-1) in estrogen receptor (ER) positive breast cancer remains to be fully elucidated. We evaluated TIMP-1 as a prognostic marker in patients treated with adjuvant tamoxifen and investigated TIMP-1s association with Ki67 and ER...... = 0.48; OR 0.68; 95% CI 0.23-1.99). Conclusion. TIMP-1 does not appear to be prognostic in breast cancer patients receiving adjuvant tamoxifen. We identified a negative association between TIMP-1 and Ki67. We did not confirm our previous in vitro findings of a negative association between TIMP-1...

  8. Aggregates in monoclonal antibody manufacturing processes. (United States)

    Vázquez-Rey, María; Lang, Dietmar A


    Monoclonal antibodies have proved to be a highly successful class of therapeutic products. Large-scale manufacturing of pharmaceutical antibodies is a complex activity that requires considerable effort in both process and analytical development. If a therapeutic protein cannot be stabilized adequately, it will lose partially or totally its therapeutic properties or even cause immunogenic reactions thus potentially further endangering the patients' health. The phenomenon of protein aggregation is a common issue that compromises the quality, safety, and efficacy of antibodies and can happen at different steps of the manufacturing process, including fermentation, purification, final formulation, and storage. Aggregate levels in drug substance and final drug product are a key factor when assessing quality attributes of the molecule, since aggregation might impact biological activity of the biopharmaceutical. In this review it is analyzed how aggregates are formed during monoclonal antibody industrial production, why they have to be removed and the manufacturing process steps that are designed to either minimize or remove aggregates in the final product. Copyright © 2011 Wiley Periodicals, Inc.


    Directory of Open Access Journals (Sweden)

    Michael Sela


    Full Text Available History of developing synergy between monoclonal antibodies, anti-tumor activity of monoclonal antibodies against tyrosine-kinases receptors EGFR/ErbB-1 and HER2/ErbB-2 as well as growth factor VEGF in various combinations are considered in the article. There were proposed hypotheses about potential molecular mechanisms underlay synergy between monoclonal antibodies (for homo- and hetero combinations of antibodies appropriately specific for antigenic determinants on the same or different receptors. Future trends in researches necessary to deeper understanding causes of this phenomenon and perspectives for practical application of monoclonal antibodies acted synergistically as immunotherapeutic drugs for human tumors treatment are reviewed.

  10. Ki-67 is an independent indicator in non-muscle invasive bladder cancer (NMIBC); combination of EORTC risk scores and Ki-67 expression could improve the risk stratification of NMIBC. (United States)

    Ding, Weihong; Gou, Yuancheng; Sun, Chuanyu; Xia, Guowei; Wang, Hong; Chen, Zhongqing; Tan, Jun; Xu, Ke; Qiang, Ding


    To prove the predicting role of Ki-67 expression and to demonstrate that the combination of European Organization for Research and Treatment of Cancer (EORTC) risk scores and Ki-67 staining status could improve the risk stratification in a large series of patients with non-muscle invasive bladder cancer (NMIBC). From October 2002 to July 2010, in our cohort, 332 patients who were treated with transurethral resection of the bladder tumor were diagnosed with NMIBC by histopathologic analysis. Two experienced uropathologists rereviewed the slides. The EORTC risk scores for recurrence and progression were determined. Ki-67 expression was evaluated using immunohistochemical studies and scored for intensity and area of staining. We correlated Ki-67 expression scores with clinical and pathologic variables. We evaluated the prognosis role of EORTC risk scores, Ki-67 staining, and their combination on tumor recurrence-free survival and progression-free survival (PFS) by univariate analysis, multivariate analysis, and Kaplan-Meier survival curves. With a median follow-up of 47 (range, 2-124) months, 119 patients (35.8%) had tumor recurrence and 40 patients (12%) had tumor progression. Ki-67 positivity (Ki-67>25%) was reported in 108 tumors (32.5%), and it was significantly associated with high EORTC risk scores for both tumor recurrence and progression. In univariate analysis, multifocality, tumor size, tumor stage, tumor grade, and Ki-67 staining correlated with recurrence-free survival, whereas tumor size, tumor stage, tumor grade, concomitant CIS, and Ki-67 staining correlated with PFS. In multivariable analysis, Ki-67 expression was an independent risk factor for predicting tumor recurrence (hazard ratio, 2.14; PEORTC risk scores and Ki-67 staining led to more accurate prediction for tumor recurrence and progression (log-rank test; PEORTC risk scores with Ki-67 expression could improve the risk stratification for both recurrence and progression in NMIBC. Copyright

  11. A novel role for junctional adhesion molecule-A in tumor proliferation: modulation by an anti-JAM-A monoclonal antibody. (United States)

    Goetsch, Liliane; Haeuw, Jean-François; Beau-Larvor, Charlotte; Gonzalez, Alexandra; Zanna, Laurence; Malissard, Martine; Lepecquet, Anne-Marie; Robert, Alain; Bailly, Christian; Broussas, Matthieu; Corvaia, Nathalie


    To identify new potential targets in oncology, functional approaches were developed using tumor cells as immunogens to select monoclonal antibodies targeting membrane receptors involved in cell proliferation. For that purpose cancer cells were injected into mice and resulting hybridomas were screened for their ability to inhibit cell proliferation in vitro. Based on this functional approach coupled to proteomic analysis, a monoclonal antibody specifically recognizing the human junctional adhesion molecule-A (JAM-A) was defined. Interestingly, compared to both normal and tumor tissues, we observed that JAM-A was mainly overexpressed on breast, lung and kidney tumor tissues. In vivo experiments demonstrated that injections of anti-JAM-A antibody resulted in a significant tumor growth inhibition of xenograft human tumors. Treatment with monoclonal antibody induced a decrease of the Ki67 expression and downregulated JAM-A levels. All together, our results show for the first time that JAM-A can interfere with tumor proliferation and suggest that JAM-A is a potential novel target in oncology. The results also demonstrate that a functional approach coupled to a robust proteomic analysis can be successful to identify new antibody target molecules that lead to promising new antibody-based therapies against cancers.

  12. High Ki-67 Immunohistochemical Reactivity Correlates With Poor Prognosis in Bladder Carcinoma (United States)

    Luo, Yihuan; Zhang, Xin; Mo, Meile; Tan, Zhong; Huang, Lanshan; Zhou, Hong; Wang, Chunqin; Wei, Fanglin; Qiu, Xiaohui; He, Rongquan; Chen, Gang


    Abstract Ki-67 is considered as one of prime biomarkers to reflect cell proliferation and immunohistochemical Ki-67 staining has been widely applied in clinical pathology. To solve the widespread controversy whether Ki-67 reactivity significantly predicts clinical prognosis of bladder carcinoma (BC), we performed a comprehensive meta-analysis by combining results from different literature. A comprehensive search was conducted in the Chinese databases of WanFang, China National Knowledge Infrastructure and Chinese VIP as well as English databases of PubMed, ISI web of science, EMBASE, Science Direct, and Wiley online library. Independent studies linking Ki-67 to cancer-specific survival (CSS), disease-free survival (DFS), overall survival (OS), progression-free survival (PFS), and recurrence-free survival (RFS) were included in our meta-analysis. With the cut-off values literature provided, hazard ratio (HR) values between the survival distributions were extracted and later combined with STATA 12.0. In total, 76 studies (n = 13,053 patients) were eligible for the meta-analysis. It was indicated in either univariate or multivariate analysis for survival that high Ki-67 reactivity significantly predicted poor prognosis. In the univariate analysis, the combined HR for CSS, DFS, OS, PFS, and RFS were 2.588 (95% confidence interval [CI]: 1.623–4.127, P < 0.001), 2.697 (95%CI: 1.874–3.883, P < 0.001), 2.649 (95%CI: 1.632–4.300, P < 0.001), 3.506 (95%CI: 2.231–5.508, P < 0.001), and 1.792 (95%CI: 1.409–2.279, P < 0.001), respectively. The pooled HR of multivariate analysis for CSS, DFS, OS, PFS, and RFS were 1.868 (95%CI: 1.343–2.597, P < 0.001), 2.626 (95%CI: 2.089–3.301, P < 0.001), 1.104 (95%CI: 1.008–1.209, P = 0.032), 1.518 (95%CI: 1.299–1.773, P < 0.001), and 1.294 (95%CI: 1.203–1.392, P < 0.001), respectively. Subgroup analysis of univariate analysis by origin showed that Ki-67 reactivity significantly

  13. Emerging monoclonal antibodies against Clostridium difficile infection. (United States)

    Péchiné, Séverine; Janoir, Claire; Collignon, Anne


    Clostridium difficile infections are characterized by a high recurrence rate despite antibiotic treatments and there is an urgent need to develop new treatments such as fecal transplantation and immonotherapy. Besides active immunotherapy with vaccines, passive immunotherapy has shown promise, especially with monoclonal antibodies. Areas covered: Herein, the authors review the different assays performed with monoclonal antibodies against C. difficile toxins and surface proteins to treat or prevent primary or recurrent episodes of C. difficile infection in animal models and in clinical trials as well. Notably, the authors lay emphasis on the phase III clinical trial (MODIFY II), which allowed bezlotoxumab to be approved by the Food and Drug Administration and the European Medicines Agency. They also review new strategies for producing single domain antibodies and nanobodies against C. difficile and new approaches to deliver them in the digestive tract. Expert opinion: Only two human Mabs against TcdA and TcdB have been tested alone or in combination in clinical trials. However, many animal model studies have provided rationale for the use of Mabs and nanobodies in C. difficile infection and pave the way for further clinical investigation.

  14. A comparison of Ki-67 counting methods in luminal Breast Cancer: The Average Method vs. the Hot Spot Method (United States)

    Jang, Min Hye; Kim, Hyun Jung; Chung, Yul Ri; Lee, Yangkyu


    In spite of the usefulness of the Ki-67 labeling index (LI) as a prognostic and predictive marker in breast cancer, its clinical application remains limited due to variability in its measurement and the absence of a standard method of interpretation. This study was designed to compare the two methods of assessing Ki-67 LI: the average method vs. the hot spot method and thus to determine which method is more appropriate in predicting prognosis of luminal/HER2-negative breast cancers. Ki-67 LIs were calculated by direct counting of three representative areas of 493 luminal/HER2-negative breast cancers using the two methods. We calculated the differences in the Ki-67 LIs (ΔKi-67) between the two methods and the ratio of the Ki-67 LIs (H/A ratio) of the two methods. In addition, we compared the performance of the Ki-67 LIs obtained by the two methods as prognostic markers. ΔKi-67 ranged from 0.01% to 33.3% and the H/A ratio ranged from 1.0 to 2.6. Based on the receiver operating characteristic curve method, the predictive powers of the KI-67 LI measured by the two methods were similar (Area under curve: hot spot method, 0.711; average method, 0.700). In multivariate analysis, high Ki-67 LI based on either method was an independent poor prognostic factor, along with high T stage and node metastasis. However, in repeated counts, the hot spot method did not consistently classify tumors into high vs. low Ki-67 LI groups. In conclusion, both the average and hot spot method of evaluating Ki-67 LI have good predictive performances for tumor recurrence in luminal/HER2-negative breast cancers. However, we recommend using the average method for the present because of its greater reproducibility. PMID:28187177

  15. Imunoexpressão do Ki-67 no carcinoma do pênis Immunoexpression of Ki-67 antigen in carcinoma of the penis

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    Sandro M. Faria


    Full Text Available Objetivo - Estudar se a expressão imunohistoquímica do Ki-67 em carcinomas epidermóides de pênis tem relação com a agressividade biológica do tumor. Material e Métodos - Em 26 pacientes portadores da neoplasia fez-se amputação parcial ou total de pênis, em 10 dos quais associou-se linfadenectomia regional por suspeita de metástases. A idade média dos pacientes foi de 61 anos e o tempo de seguimento pós-operatório de 5± 3,2 anos. As peças operatórias conservadas em formol ou parafina foram processadas para marcação imunohistoquímica para o Ki-67 pelo método da avidina-biotina-peroxidase. Resultados - Em 19/26 (73% dos casos a marcação foi positiva, sendo 13/20 tumores bem diferenciados e 6/6 moderadamente diferenciados. Em 10/16 pacientes com doença localizada o marcador foi positivo, enquanto que essa proporção para os casos de metástases foi 9/10. A sobrevida de 5 anos foi 15/26 (65,4%. A análise estatística não mostrou correlação entre a positividade imunohistoquímica com o grau e estádio da neoplasia, nem com a sobrevida dos pacientes. Conclusão - Não se encontrou relação entre a marcação imunohistoquímica do Ki-67 com a agressividade biológica do tumor.

  16. Vitamin D Receptor, Retinoid X Receptor, Ki-67, Survivin, and Ezrin Expression in Canine Osteosarcoma

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    John Davies


    Full Text Available Canine osteosarcoma (OS is an aggressive malignant bone tumor. Prognosis is primarily determined by clinical parameters. Vitamin D has been postulated as a novel therapeutic option for many malignancies. Upon activation, vitamin D receptors (VDRs combine with retinoid receptor (RXR forming a heterodimer initiating a cascade of events. Vitamin D's antineoplastic activity and its mechanism of action in OS remain to be clearly established. Expression of VDR, RXR, Ki-67, survivin, and ezrin was studied in 33 archived, canine OS specimens. VDR, RXR, survivin, and ezrin were expressed in the majority of cases. There was no statistically significant difference in VDR expression in relationship with tumor grade, type, or locations or animal breed, age, and/or sex. No significant association (p=0.316 between tumor grade and Ki-67 expression was found; in particular, no difference in Ki-67 expression between grades 2 and 3 OSs was found, while a negative correlation was noted between Ki-67 and VDR expression (ρ=−0.466, a positive correlation between survivin and RXR expression was found (p=0.374. A significant relationship exists between VDR and RXR expression in OSs and proliferative/apoptosis markers. These results establish a foundation for elucidating mechanisms by which vitamin D induces antineoplastic activity in OS.

  17. Folate receptor and Ki-67 nucleoprotein expressions in cervical cancer tissue and their correlation

    Institute of Scientific and Technical Information of China (English)

    Ran Yan; Feng Li


    Objective:To detect the expression of both FR-α protein and ki-67 in cervical cancer tissues, and discuss the relationship between them and clinical significance.Methods:Using immunohistochemical method test normal cervical tissue and cervical cancer tissue before FR-α protein expression and the expression of Ki-67.Results:FR- protein expression in normal cervical tissues was positive for 7.0% while in cervical cancer tissue the positive rate was 82.1%. The difference was statistically significant. Ki-67 protein expression in normal cervical tissues was 0% while in cervical cancer tissue the positive rate was 80.2%. The difference was statistically significant. The two protein expression in cervical cancer stageⅠ,Ⅱ and stageⅢ were different, but the difference was not statistically significant. In cervical cancer tissues, both the two protein were positively correlated. There are correlations between them. Difference was statistically significant.Conclusion:FR-α elevated protein expression is involved in the pathogenesis of cervical cancer. FR-α protein expression in cervical cancer and precancerous tissue has correlation with Ki-67, FR-α protein maybe participate in the occurrence and development of the cell proliferation in cervical cancer.

  18. Ki-67 acts as a biological surfactant to disperse mitotic chromosomes. (United States)

    Cuylen, Sara; Blaukopf, Claudia; Politi, Antonio Z; Müller-Reichert, Thomas; Neumann, Beate; Poser, Ina; Ellenberg, Jan; Hyman, Anthony A; Gerlich, Daniel W


    Eukaryotic genomes are partitioned into chromosomes that form compact and spatially well-separated mechanical bodies during mitosis. This enables chromosomes to move independently of each other for segregation of precisely one copy of the genome to each of the nascent daughter cells. Despite insights into the spatial organization of mitotic chromosomes and the discovery of proteins at the chromosome surface, the molecular and biophysical bases of mitotic chromosome structural individuality have remained unclear. Here we report that the proliferation marker protein Ki-67 (encoded by the MKI67 gene), a component of the mitotic chromosome periphery, prevents chromosomes from collapsing into a single chromatin mass after nuclear envelope disassembly, thus enabling independent chromosome motility and efficient interactions with the mitotic spindle. The chromosome separation function of human Ki-67 is not confined within a specific protein domain, but correlates with size and net charge of truncation mutants that apparently lack secondary structure. This suggests that Ki-67 forms a steric and electrostatic charge barrier, similar to surface-active agents (surfactants) that disperse particles or phase-separated liquid droplets in solvents. Fluorescence correlation spectroscopy showed a high surface density of Ki-67 and dual-colour labelling of both protein termini revealed an extended molecular conformation, indicating brush-like arrangements that are characteristic of polymeric surfactants. Our study thus elucidates a biomechanical role of the mitotic chromosome periphery in mammalian cells and suggests that natural proteins can function as surfactants in intracellular compartmentalization.

  19. Screening for cervical cancer precursors with p16/Ki-67 dual-stained cytology

    DEFF Research Database (Denmark)

    Ikenberg, Hans; Bergeron, Christine; Schmidt, Dietmar


    Pap cytology is known to be more specific but less sensitive than testing for human papillomavirus (HPV) for the detection of high-grade cervical intraepithelial neoplasia (CIN2+). We assessed whether p16/Ki-67 dual-stained cytology, a biomarker combination indicative of transforming HPV infections...

  20. Analysis on the relation of pterygium with VEGF,SDF-1,Ki-67,PCNA and Survivin

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    Ying Song


    Full Text Available AIM:To analyze and study the relation of pterygium with vascular endothelial growth factor(VEGF,stroma cell-derived factor 1(SDF-1,tumor proliferating antigen(Ki-67,proliferating cell nuclear antigen(PCNAand survivin. METHODS:Seventy-nine patients(106 eyeswith pterygium from January 2013 to May 2015 in our hospital were selected as observation group. Seventy-nine persons with normal conjunctiva during the same period were selected as control group. Then the number of positive cells and staining intensity classification of the two groups for VEGF,SDF-1,Ki-67,PCNA and survivin were compared,and the detection results of patients with different gender,stages and types were compared too. Then the relation between pterygium and those indexes were analyzed by the Logistic analysis. RESULTS:The number of positive cells and staining intensity classification of observation group for VEGF,SDF-1,Ki-67,PCNA and survivin were all higher than those of control group,and the detection results of patients with different stages and types had certain differences too(all PP>0.05. All those indexes had close relation to pterygium by the Logistic analysis. CONCLUSION:The expression of VEGF,SDF-1,Ki-67,PCNA and survivin in tissue of patients with pterygium all show abnormal state,and those indexes all have close relation to pterygium.

  1. P63 and Ki-67 Expression in Dentigerous Cyst and Ameloblastomas

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    Zohreh Jaafari-Ashkavandi


    Full Text Available Statement of the Problem: P63 gene is a member of TP53 and its homologous gene family. Its expression was observed in some odontogenic lesions, more expression in aggressive lesions. Purpose: This study aimed to investigate the possible diagnostic impact of P63 protein on dentigerous cysts and various types of ameloblastoma. Its expression with Ki-67 proliferation marker was also compared. Materials and Method: This cross-sectional retrospective study was enrolled on 25 cases of dentigerous cyst including 21 unicystic ameloblastomas and 17 conventional ameloblastomas. The expression of P63 and Ki-67 was assessed by immunohistochemical (IHC examinations. Data were analyzed by employing Mann-Whitney and correlation coefficient tests. Results: P63 expression was significantly higher in ameloblastoma than unicystic ameloblastoma and dentigerous cysts. There was no significant difference between unicystic ameloblastoma and dentigerous cyst in P63 expression. A 90% cut-off point was obtained for basal layer which gave 88% sensitivity and 78% specificity to distinguish more invasive lesions from others. There was not any correlation between P63 and Ki-67 immunostaining in the three study groups. Conclusion: More aggressiveness and more invasiveness of odontogenic lesions depicted higher rate and also more intensive expression of P63. Moreover, the expression of P63 protein had not any correlation with Ki-67 protein in dentigerous cysts and ameloblastomas.

  2. Anaphylaxis to chemotherapy and monoclonal antibodies. (United States)

    Castells, Mariana C


    Hypersensitivity reactions are increasingly prevalent, although underrecognized and underreported. Platins induce immunoglobulin E-mediated sensitization; taxenes and some monoclonal antibodies can induce reactions at first exposure. Severe hypersensitivity can preclude first-line therapy. Tryptase level at the time of a reaction is a useful diagnostic tool. Skin testing provides a specific diagnosis. Newer tests are promising diagnostic tools to help identify patients at risk before first exposure. Safe management includes rapid drug desensitization. This review provides information regarding the scope of hypersensitivity and anaphylactic reactions induced by chemotherapy and biological drugs, as well as diagnosis, management, and treatment options.

  3. The Role of Monoclonal Antibodies in the Management of Leukemia (United States)

    Al-Ameri, Ali; Cherry, Mohamad; Al-Kali, Aref; Ferrajoli, Alessandra


    This article will review the monoclonal antibodies more commonly used in leukemias. In the last three decades, scientists have made considerable progress understanding the structure and the functions of various surface antigens, such as CD20, CD33. The introduction of rituximab, an anti CD20 monoclonal antibody, had a great impact in the treatment of lymphoproliferative disorders. Gemtuzumab, an anti CD 33 conjugated monoclonal antibody has activity in acute mylegenous leukemia (AML). As this field is undergoing a rapid growth, the years will see an increasing use of monoclonal antibodies in hematological malignancies.

  4. The Role of Monoclonal Antibodies in the Management of Leukemia

    Directory of Open Access Journals (Sweden)

    Mohamad Cherry


    Full Text Available This article will review the monoclonal antibodies more commonly used in leukemias. In the last three decades, scientists have made considerable progress understanding the structure and the functions of various surface antigens, such as CD20, CD33. The introduction of rituximab, an anti CD20 monoclonal antibody, had a great impact in the treatment of lymphoproliferative disorders. Gemtuzumab, an anti CD 33 conjugated monoclonal antibody has activity in acute mylegenous leukemia (AML. As this field is undergoing a rapid growth, the years will see an increasing use of monoclonal antibodies in hematological malignancies.

  5. Induction and characterization of monoclonal anti-idiotypic antibodies reactive with idiotopes of canine parvovirus neutralizing monoclonal antibodies.

    NARCIS (Netherlands)

    G.F. Rimmelzwaan (Guus); J. van Es (Johan); G.A. Drost; F.G.C.M. Uytdehaag (Fons); A.D.M.E. Osterhaus (Albert)


    textabstractMonoclonal anti-idiotypic (anti-Id) antibodies (Ab2) were generated against idiotypes (Id) of canine parvovirus (CPV) specific monoclonal antibodies (MoAbs). The binding of most of these anti-Id antibodies to their corresponding Id could be inhibited by antigen, thus classifying these an

  6. Immunohistochemical expression of GLUT-1 and Ki-67 in chronic plaque psoriasis. (United States)

    Abdou, Asmaa G; Maraee, Alaa H; Eltahmoudy, Mohamed; El-Aziz, Reem A


    Many inflammatory mediators and other biological markers are upregulated in psoriatic lesions; some of these alterations also persist in nonlesional skin. Glucose is the major source of energy for cells, and glucose transporter 1 is the most common glucose transporter in humans (GLUT-1). The present study aimed at evaluating the pattern of expression of GLUT-1 and Ki-67 in psoriatic skin (involved and uninvolved) and correlating their expression with the clinicopathological parameters in the studied patients. This study was carried out on 30 patients presented with chronic plaque psoriasis and 10 apparently healthy volunteers as a control group. GLUT-1 was not expressed in epidermis of normal skin, whereas it was expressed in 76.6% of uninvolved and 86.7% of involved skin of psoriatic patients, where both the latter differed significantly regarding the intensity (P = 0.001) and localization (P = 0.001) of GLUT-1 expression. The percentage of Ki-67 expression did not differ significantly between involved and uninvolved skin of psoriatic patients, but they were higher than that of normal skin of control group. Nucleolar pattern of Ki-67 expression was significantly associated with male sex (P = 0.05), marked parakeratosis (P = 0.01), and marked angiogenesis (P = 0.05). GLUT-1 expression was associated with degree of acanthosis and percentage of Ki-67 expression. From this study, GLUT-1 is upregulated in psoriatic epidermis and may be involved in facilitation of keratinocyte proliferation in psoriasis. Nucleolar pattern of Ki-67 is an indicator of progressive keratinocyte proliferation in psoriasis.

  7. Monoclonal antibody disulfide reduction during manufacturing (United States)

    Hutterer, Katariina M.; Hong, Robert W.; Lull, Jonathon; Zhao, Xiaoyang; Wang, Tian; Pei, Rex; Le, M. Eleanor; Borisov, Oleg; Piper, Rob; Liu, Yaoqing Diana; Petty, Krista; Apostol, Izydor; Flynn, Gregory C.


    Manufacturing-induced disulfide reduction has recently been reported for monoclonal human immunoglobulin gamma (IgG) antibodies, a widely used modality in the biopharmaceutical industry. This effect has been tied to components of the intracellular thioredoxin reduction system that are released upon cell breakage. Here, we describe the effect of process parameters and intrinsic molecule properties on the extent of reduction. Material taken from cell cultures at the end of production displayed large variations in the extent of antibody reduction between different products, including no reduction, when subjected to the same reduction-promoting harvest conditions. Additionally, in a reconstituted model in which process variables could be isolated from product properties, we found that antibody reduction was dependent on the cell line (clone) and cell culture process. A bench-scale model using a thioredoxin/thioredoxin reductase regeneration system revealed that reduction susceptibility depended on not only antibody class but also light chain type; the model further demonstrates that the trend in reducibility was identical to DTT reduction sensitivity following the order IgG1λ > IgG1κ > IgG2λ > IgG2κ. Thus, both product attributes and process parameters contribute to the extent of antibody reduction during production. PMID:23751615

  8. Predictive value of Ki67 for adjuvant chemotherapy in node-negative, hormone receptor-positive breast cancer. (United States)

    Sutepvarnon, Apisada; Warnnissorn, Malee; Srimuninnimit, Vichien


    Ki67 labeling index (Ki67 LI) is a measure of tumor proliferation. In breast cancer, evidence supporting its prognostic value is clear and its predictive value for response to treatment finds some benefits. However studies of Ki67 LI as a predictive marker in early breast cancer are still limited worldwide and there is no data in Thailand. To assess the predictive value of Ki67 expression for adjuvant chemotherapy in patients with node-negative, hormone receptor-positive breast cancer The authors retrospectively evaluated 127 diagnosed early breast cancer with node-negative, hormone receptor-positive patients and receiving adjuvant systemic treatment at Siriraj hospital. Disease free survival (DFS) was compared with the log-rank test according to Ki67 LI and adjuvant systemic treatment (chemoendocrine therapy and endocrine therapy alone). At a median follow-up of 3.3 years. The 5-year DFS rate was 79% for patients with low Ki67 expression and 75% for patients with high Ki67 expression. Of the 127 patients, 56 (44.1%) received chemoendocrine therapy and 71 (55.9%) were treated with endocrine therapy alone. There was no different effect of DFS among those receiving adjuvant endocrine therapy alone and those receiving adjuvant chemoendocrine therapy depending on Ki67 expression. Among patients with node-negative, hormone receptor-positive breast cancer, a high Ki67 LI had worse DFS trend than a low Ki67 LI but the Ki67 LI did not predict the efficacy of adjuvant chemotherapy.

  9. Clinicopathological and Prognostic Value of Ki-67 Expression in Bladder Cancer: A Systematic Review and Meta-Analysis.

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    Yuejun Tian

    Full Text Available Ki-67 is an established marker of cell proliferation, and the Ki-67 index correlates with the clinical course of several cancer types, including bladder cancer (BC. However, the clinicopathological and prognostic significance of Ki-67 in bladder cancer remains unclear. Therefore, we performed a systematic review and meta-analysis to clarify this relationship.A comprehensive literature search for relevant studies published up to February 1, 2016, was performed using PubMed, Cochrane Library, Embase and ISI Web of Knowledge. The effects of Ki-67 expression on survival outcome in patients with BC and BC subtypes were evaluated. Furthermore, the relationship between Ki-67 expression and the clinicopathological features of BC were assessed.Thirty-one studies with 5147 bladder cancer patients were selected for evaluation. Ki-67 expression was significantly associated with shorter recurrence-free (HR 1.69, 95% CI: 1.33-2.14, progression-free (HR 1.89, 95% CI: 1.43-2.51, overall (HR 2.03, 95% CI: 1.31-3.16, and cancer-specific (HR 1.69, 95% CI: 1.47-1.95 survival. Moreover, whereas high expression was more common in high tumor stage, recurrence status, tumor size, there was no correlation between high Ki-67 expression and age, gender, smoking habits, and tumor number. Importantly, analysis of the different subgroups of BC suggested that significant correlations between high Ki-67 expression and survival outcome (recurrence-free/progression-free/overall/cancer-specific survival are present only in European-American patients.The present results indicate that over-expression of Ki-67 is distinctly correlated with poor patient survival. Ki-67 may serve as a valuable biomarker for prognosis in BC patients, particularly in non-Asian BC patients. The results suggest no significant association between Ki-67 expression and BC prognosis in Asian patients. Further efforts are needed to fully clarify this relationship.

  10. Generation of monoclonal antibodies to native active human glycosyltransferases

    DEFF Research Database (Denmark)

    Vester-Christensen, Malene Bech; Bennett, Eric Paul; Clausen, Henrik;


    using monoclonal antibodies therefore provides an excellent strategy to analyze the glycosylation process in cells. A major drawback has been difficulties in generating antibodies to glycosyltransferases and validating their specificities. Here we describe a simple strategy for generating...

  11. Expression of p53 and Ki-67 in the Gliomas and its Clinical Significance%Ki-67、p53在脑胶质瘤中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    周开甲; 张鸣; 刘伯伟


    目的 探讨Ki-67和p53与脑胶质瘤分级的关系.方法 采用免疫组化SP法检测60例脑胶质瘤标本和10例正常脑组织标本Ki-67和p53的表达.结果 Ki-67和p53在正常脑组织与脑胶质瘤中的表达差异均有统计学意义(Ki-67:χ2=13.081,P=0.004;p53:χ2=11.667,P=0.009);Ⅰ~Ⅱ级胶质细胞瘤与高度恶性Ⅲ~Ⅳ级胶质细胞瘤Ki-67和p53的表达差异均有统计学意义(Ki-67:χ2=12.589,P=0.006;p53:χ2=12.721,P=0.005).结论 脑胶质瘤中Ki-67及p53的阳性表达率与其恶性度相关,均可作为判断胶质瘤恶性程度的指标.%Objective To determine the relationship between the expressions of Ki-67 and p53 and the grading of the gliomas. Methods The expressions of Ki-67 and p53 in 60 samples of human gliomas and 10 normal tissue specimens were de-tected by the immunohistochemistry method ( SP ). Results There's significant diference in the expression of T Ki-67 and p53 between normal brain tissue and glioma ( Ki-67 :x2 = 13. 081, P =0. 004 ; p53 : x2 = 11. 667, P = 0. 009 ). Significant difference exist between grade Ⅰ~Ⅱ glioma and grade Ⅲ~Ⅳ glioma( Ki-67 : x2 = 12. 589, P = 0. 006; p53 : x2 = 12. 121, P = 0. 005 ). Conclusion Ki-67 and p53 express in high percentage of gliomas. The expressions of Ki-67 and p53 in gliomas may be used as tumor markers evaluating the histograde of human glioma.

  12. Ki-67 and p53 correlation prognostic value in squamous cell carcinomas of the oral cavity and tongue Valor prognóstico da correlação do Ki-67 e p53 em carcinomas epidermoides da cavidade oral e língua

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    Rafael Da Ros Motta


    Full Text Available Epidermoid carcinomas represent from 90% to 95% of oral cavity malignant neoplasias, making up 13,470 cases/year. AIMS: To correlate p53 and Ki-67 expressions in mouth and tongue carcinomas with lymph node status, gender, histological grade, tumor volume and pathological stage. MATERIALS AND METHODS: We carried out a retrospective study of 28 cases of mouth and tongue epidermoid carcinomas. They were submitted to immunohistochemical study in order to check the expression of p53 and Ki-67 antibodies and statistically compare them in terms of lymph node status, gender, histological grade, tumor volume and pathological staging. RESULTS: The individually analyzed p53 proved to have statistical significance (pO carcinoma epidermoide representa 90% a 95% das neoplasias malignas da cavidade oral, responsável por 13.470 casos/ano. OBJETIVOS: Correlacionar a expressão do p53 e Ki-67 nos carcinomas epidermoides de cavidade oral e língua com o estado linfonodal, sexo, grau histológico, volume tumoral e estadiamento patológico. MATERIAIS E MÉTODOS: Foi realizado estudo retrospectivo de 28 casos de carcinomas epidermoides da cavidade oral e língua. Estes foram submetidos à técnica de imunoistoquímica para verificar a expressão dos anticorpos p53 e Ki-67 e compará-los estatisticamente quanto ao status linfonodal, sexo, grau histológico, volume tumoral e estadiamento patológico. RESULTADOS: O p53 analisado individualmente mostrou significância estatística (p<0,05 quando comparado com o volume tumoral (p=0,029. Apesar de uma forte tendência, a relação de p53 com estado linfonodal não foi significativa. Quando o p53 + Ki67 foram analisados, o volume tumoral mostrou p < 0,05 (p = 0,029. DISCUSSÃO: A literatura mostra que a expressão dos marcadores p53 e Ki-67 está relacionada com presença de metástases para linfonodos e pior prognóstico. CONCLUSÃO: Nos carcinomas epidermoides da cavidade oral e língua o p53 e o Ki-67 est

  13. A monoclonal antibody toolkit for C. elegans.

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    Gayla Hadwiger

    Full Text Available BACKGROUND: Antibodies are critical tools in many avenues of biological research. Though antibodies can be produced in the research laboratory setting, most research labs working with vertebrates avail themselves of the wide array of commercially available reagents. By contrast, few such reagents are available for work with model organisms. METHODOLOGY/PRINCIPAL FINDINGS: We report the production of monoclonal antibodies directed against a wide range of proteins that label specific subcellular and cellular components, and macromolecular complexes. Antibodies were made to synaptobrevin (SNB-1, a component of synaptic vesicles; to Rim (UNC-10, a protein localized to synaptic active zones; to transforming acidic coiled-coil protein (TAC-1, a component of centrosomes; to CENP-C (HCP-4, which in worms labels the entire length of their holocentric chromosomes; to ORC2 (ORC-2, a subunit of the DNA origin replication complex; to the nucleolar phosphoprotein NOPP140 (DAO-5; to the nuclear envelope protein lamin (LMN-1; to EHD1 (RME-1 a marker for recycling endosomes; to caveolin (CAV-1, a marker for caveolae; to the cytochrome P450 (CYP-33E1, a resident of the endoplasmic reticulum; to beta-1,3-glucuronyltransferase (SQV-8 that labels the Golgi; to a chaperonin (HSP-60 targeted to mitochondria; to LAMP (LMP-1, a resident protein of lysosomes; to the alpha subunit of the 20S subcomplex (PAS-7 of the 26S proteasome; to dynamin (DYN-1 and to the alpha-subunit of the adaptor complex 2 (APA-2 as markers for sites of clathrin-mediated endocytosis; to the MAGUK, protein disks large (DLG-1 and cadherin (HMR-1, both of which label adherens junctions; to a cytoskeletal linker of the ezrin-radixin-moesin family (ERM-1, which localized to apical membranes; to an ERBIN family protein (LET-413 which localizes to the basolateral membrane of epithelial cells and to an adhesion molecule (SAX-7 which localizes to the plasma membrane at cell-cell contacts. In addition to

  14. Automated characterization and counting of Ki-67 protein for breast cancer prognosis: A quantitative immunohistochemistry approach. (United States)

    Mungle, Tushar; Tewary, Suman; Arun, Indu; Basak, Bijan; Agarwal, Sanjit; Ahmed, Rosina; Chatterjee, Sanjoy; Maity, Asok Kumar; Chakraborty, Chandan


    Ki-67 protein expression plays an important role in predicting the proliferative status of tumour cells and deciding the future course of therapy in breast cancer. Immunohistochemical (IHC) determination of Ki-67 score or labelling index, by estimating the fraction of Ki67 positively stained tumour cells, is the most widely practiced method to assess tumour proliferation (Dowsett et al. 2011). Accurate manual counting of these cells (specifically nuclei) due to complex and dense distribution of cells, therefore, becomes critical and presents a major challenge to pathologists. In this paper, we suggest a hybrid clustering algorithm to quantify the proliferative index of breast cancer cells based on automated counting of Ki-67 nuclei. The proposed methodology initially pre-processes the IHC images of Ki-67 stained slides of breast cancer. The RGB images are converted to grey, L*a*b*, HSI, YCbCr, YIQ and XYZ colour space. All the stained cells are then characterized by two stage segmentation process. Fuzzy C-means quantifies all the stained cells as one cluster. The blue channel of the first stage output is given as input to k-means algorithm, which provides separate cluster for Ki-67 positive and negative cells. The count of positive and negative nuclei is used to calculate the F-measure for each colour space. A comparative study of our work with the expert opinion is studied to evaluate the error rate. The positive and negative nuclei detection results for all colour spaces are compared with the ground truth for validation and F-measure is calculated. The F-measure for L*a*b* colour space (0.8847) provides the best statistical result as compared to grey, HSI, YCbCr, YIQ and XYZ colour space. Further, a study is carried out to count nuclei manually and automatically from the proposed algorithm with an average error rate of 6.84% which is significant. The study provides an automated count of positive and negative nuclei using L*a*b*colour space and hybrid

  15. HPV-DNA testing and Ki-67 immunocytochemistry in liquid based cervical cytology in prostitute women

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    Esin ATİK


    Full Text Available HPV causes several changes in the function of host genes, and these interactions cause deregulation of the cell cycle manifested by abnormal expression of cell cycle associated proteins, such as Ki-67. The detection of Ki-67 can play a role in screening and diagnosis of HPV infection with risk of progression towards dysplasia and carcinoma. To show this relation in prostitute women, cervical cells were collected in the PapSpin Collection Fluid. A starting volume of 1000 µl for each sample, and a 200 µl cell suspension were used to prepare each sample for thin layer liquid based cytology and then they were stained by Papanicolaou method. The cytological results were classified according to the Bethesda 2001 system. From the remaining cell suspension of 800 µl, a 400 µl sample was used for HPV-DNA detection by PCR, a 50 µl alliquot was used to make thin layer preparations for immunocytochemistry. Single antigen staining was performed with Ki-67 protein. Cells were considered immunopositive if the nuclei were stained. All cells in one high power field (x400 were counted, and the fraction of immunopositive cells on the slide was calculated. This fraction was expressed as the number of positive cells per 1000 cells to facilitate comparisons of differential cell counts. HPV types 6 and 32 in the study, and HPV types 6 and 51 in the control group were detected. The mean Ki-67 values were 2.7±1.2 and 3.6±4.1 in HPV positive and negative cases respectively. There was a positive correlation only with nuclear changes and HPV positivity (x2=28.8, p<0.001. There was not any significant correlation between HPV or Ki-67 and leukocytosis. An association with HPV and contraception, smoking, and concurrent genital infection was not found. The prevalence of HPV types in different geographical locations and races may indicate different etiologies of cervical cancer. Our results suggest that Ki-67 immunocytochemistry is not useful as a surrogate marker for

  16. Increased proliferation activity measured by immunoreactive Ki67 is associated with survival improvement in rectal/recto sigmoid cancer

    Institute of Scientific and Technical Information of China (English)

    Eeva Salminen; Salla Palmu; Tero Vahlberg; Peter J. Roberts; Karl-Owe S(o)derstr(o)m


    AIM: To assess the expression of Ki67 as prognosticator in rectal/recto sigmoid cancer.METHODS: Samples from 146 patients with rectal and recto sigmoid cancer were studied for expression of Ki67 and its prognostic significance in comparison with clinicopathological predictors of survival. Formalin-fixed, paraffin-embedded tissues from 6 (4.1%) patients with T1, 26 (17.8%) with T2, 94 (64.4%) with T3, and 20 (13.7%) with T4 tumors were studied. Ki67 expression was determined immunohistochemically. Samples were divided according to mean value into high (>40%) and low (≤40%) expression. Areas of extensive proliferation (>50%) were defined as 'hot spot' areas. RESULTS: Hot spot areas were present in samples regardless of histopathological grade. Lower TNM and Dukes stage and higher expression of Ki67 and presence of Ki67 hot spot areas in histopathological samples were associated with better survival, whereas no association was observed with histopathological grade (P = 0.78). In Cox multivariate regression analysis, significant prognostic factors were Dukes stage (P<0.001), presence of lymph node metastases (P = 0.015), age (P = 0.035) andpresence of Ki67 hot spot areas (P = 0.044). CONCLUSION: Proliferative activity as measured by Ki67 in rectal cancer is associated with survival improvement compared with patients with low Ki67. Areas of prognostically significant increased proliferation were found independently of histopathological tumor grade.

  17. Immunochemical Characterization of Anti-Acetylcholinesterase Inhibitory Monoclonal Antibodies (United States)


    formation. This conformation was first proposed using studies with monoclonal antibodies against a synthetic peptide mimicking the sequence of the...distinct antigenic determinants on dengue -2 virus using monoclonal antibodies, Am. J. Trop. Med. Hyg., 31 (1982) 548-555. 7 D. De la Hoz, B.P. Doctor

  18. Monoclonal Antibodies Specific for Hippurate Hydrolase of Campylobacter jejuni


    Steele, Marina; Gyles, Carlton; Chan, Voon Loong; Odumeru, Joseph


    Eleven monoclonal antibodies raised against recombinant Campylobacter jejuni hippurate hydrolase were tested for binding to lysates from 19 C. jejuni strains, 12 other Campylobacter strains, and 21 non-Campylobacter strains. Several monoclonal antibodies bound to C. jejuni but not to other Campylobacter species and may be useful in a species-specific immunoassay.

  19. Immunoexpression of Ki-67, MCM2, and MCM3 in Ameloblastoma and Ameloblastic Carcinoma and Their Correlations with Clinical and Histopathological Patterns. (United States)

    Carreón-Burciaga, Ramón Gil; González-González, Rogelio; Molina-Frechero, Nelly; Bologna-Molina, Ronell


    Cell proliferation assays are performed using antibodies against nuclear proteins associated with DNA replication. These nuclear proteins have gained special interest to predict the biological and clinical behaviors of various tumors. The aim of this study was to analyze the presence of Ki-67 protein and the minichromosome maintenance-2 (MCM2) and maintenance-3 (MCM3) proteins in ameloblastoma. Materials and Methods. Cell proliferation marker expression levels were assessed via immunohistochemistry in 111 ameloblastoma cases (72 unicystic ameloblastoma samples, 38 solid/multicystic ameloblastoma samples, and 1 ameloblastic carcinoma). The label index was performed as described previously. Results. MCM2 and MCM3 showed higher proliferation indexes in all variants of ameloblastoma compared to the classic marker Ki-67. No correlation between the proliferation index and the clinical and protein expression data was observed. Conclusion. The results suggest that clinical features do not directly affect tumor cell proliferation. Moreover, the high levels of cellular proliferation of MCM2 and MCM3 compared with Ki-67 may indicate that MCM2 and MCM3 are more sensitive markers for predicting the growth rate and eventually might be helpful as a tool for predicting aggressive and recurrent behaviors in these tumors.

  20. Development of syngeneic monoclonal anti-idiotype antibodies to mouse monoclonal anti-asialoglycoprotein receptor antibody.

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    Hirai M


    Full Text Available Anti-idiotype antibodies (Ab2 play an important role in the homeostasis of immune responses and are related to the development and the disease activity of certain autoimmune diseases. The asialoglycoprotein receptor (ASGPR is considered one of the target antigens in the pathogenesis of autoimmune chronic active hepatitis (AIH. We previously developed a mouse monoclonal antibody (clone 8D7 which recognizes rat and human ASGPR. In this study, to help investigate the anti-ASGPR antibody-anti-idiotype antibody network in patients with AIH, we developed a syngeneic mouse monoclonal Ab2 to the 8D7 anti-ASGPR antibody (Ab1. One clone, designated as 3C8, tested positive for specific reactivity to 8D7-Ab1 and did not bind to other irrelevant immunoglobulins. By competitive inhibition assays, the binding of 8D7-Ab1 to liver membrane extracts, i.e., the crude antigen preparation, was inhibited by 3C8-Ab2 in a dose-dependent manner, and the binding of 8D7-Ab1 to 3C8-Ab2 was inhibited by the liver membrane extracts. In the immunohistochemical analysis, 3C8-Ab2 blocked the specific staining of sinusoidal margins of rat hepatocytes by 8D7-Ab1. These results suggest that 3C8 anti-idiotype antibody recognizes the specific idiotypic determinants within the antigen-binding site of 8D7-Ab1.

  1. Monoclonal antibodies based on hybridoma technology. (United States)

    Yagami, Hisanori; Kato, Hiroshi; Tsumoto, Kanta; Tomita, Masahiro


    Based on the size and scope of the present global market for medicine, monoclonal antibodies (mAbs) have a very promising future, with applications for cancers through autoimmune ailments to infectious disease. Since mAbs recognize only their target antigens and not other unrelated proteins, pinpoint medical treatment is possible. Global demand is dramatically expanding. Hybridoma technology, which allows production of mAbs directed against antigens of interest is therefore privileged. However, there are some pivotal points for further development to generate therapeutic antibodies. One is selective generation of human mAbs. Employment of transgenic mice producing human antibodies would overcome this problem. Another focus is recognition sites and conformational epitopes in antigens may be just as important as linear epitopes, especially when membrane proteins such as receptors are targeted. Recognition of intact structures is of critical importance for medical purposes. In this review, we describe patent related information for therapeutic mAbs based on hybridoma technology and also discuss new advances in hybridoma technology that facilitate selective production of stereospecific mAbs.

  2. Monoclonal Antibodies Against Xenopus Greatwall Kinase (United States)

    Wang, Ling; Fisher, Laura A.; Wahl, James K.


    Mitosis is known to be regulated by protein kinases, including MPF, Plk1, Aurora kinases, and so on, which become active in M-phase and phosphorylate a wide range of substrates to control multiple aspects of mitotic entry, progression, and exit. Mechanistic investigations of these kinases not only provide key insights into cell cycle regulation, but also hold great promise for cancer therapy. Recent studies, largely in Xenopus, characterized a new mitotic kinase named Greatwall (Gwl) that plays essential roles in both mitotic entry and maintenance. In this study, we generated a panel of mouse monoclonal antibodies (MAbs) specific for Xenopus Gwl and characterized these antibodies for their utility in immunoblotting, immunoprecipitation, and immunodepletion in Xenopus egg extracts. Importantly, we generated an MAb that is capable of neutralizing endogenous Gwl. The addition of this antibody into M-phase extracts results in loss of mitotic phosphorylation of Gwl, Plk1, and Cdk1 substrates. These results illustrate a new tool to study loss-of-function of Gwl, and support its essential role in mitosis. Finally, we demonstrated the usefulness of the MAb against human Gwl/MASTL. PMID:22008075

  3. Ki-67 and CEA expression as prognostic markers in Dukes' C colorectal cancer. (United States)

    Ishida, Hideyuki; Miwa, Hideaki; Tatsuta, Masayuki; Masutani, Seizo; Imamura, Hiroshi; Shimizu, Junzo; Ezumi, Koji; Kato, Hitoshi; Kawasaki, Takatoshi; Furukawa, Hiroshi; Kawakami, Hitoshi


    This study investigated the relationship between clinicopathological or immunohistochemical factors and postoperative prognosis for Dukes' C colorectal cancer. Short-term survivors died from cancer within 2 years of surgery, whereas long-term survivors were disease-free for over 10 years. The groups differed in Ki-67 antigen and CEA expression in colon cancer, and CEA expression in rectal cancer that was limited to the metastatic lymph nodes. The immunohistochemical scores were higher in short-term survivors. Our data suggest that the characteristics of metastatic lymph nodes are important as a predictor of the aggressiveness of tumor behavior and that the expression of Ki-67 antigen or CEA there may be a useful indicators of patients' survival in Dukes' C colorectal cancer.

  4. Chromogranin A, Ki-67 index and IGF-related genes in patients with neuroendocrine tumors (United States)

    van Adrichem, R C S; Hofland, L J; Feelders, R A; De Martino, M C; van Koetsveld, P M; van Eijck, C H J; de Krijger, R R; Sprij-Mooij, D M; Janssen, J A M J L; de Herder, W W


    Chromogranin A (CgA) and the Ki-67 proliferation index are considered as important biochemical and pathological markers for clinical behaviour of gastroenteropancreatic neuroendocrine tumors (GEP NETs), respectively. The IGF system has been suggested as an important regulator of GEP NET proliferation and differentiation. A possible relationship between serum CgA (sCgA), Ki-67 proliferation index, and expression of IGF-related genes in patients with GEP NETs has not been demonstrated yet. This study investigates the relationship between sCgA, the Ki-67 proliferation index, and the expression of IGF-related genes in GEP NET tissues and their relation with 5-year survival. Tumor and blood samples from 22 GEP NET patients were studied. Tumoral mRNA expression of IGF-related genes (IGFs: IGF1, IGF2; IGF receptors: IGF1R, IGF2R; insulin receptors: subtype A (IR-A) and B (IR-B); IGF-binding proteins (IGFBPs): IGFBP1, IGFBP2, IGFBP3, and IGFBP6) was measured using quantitative RT-PCR. Ki-67 proliferation index was determined using immunohistochemistry. sCgA was measured with ELISA. Five-year survival in patients with nonelevated sCgA (n=11) was 91 vs 46% in patients with elevated sCgA (n=11) (P=0.006). IR-A mRNA expression was significantly higher in tumors obtained from patients with elevated sCgA than in those from patients with nonelevated sCgA (6.42±2.08 vs 2.60±0.40; P=0.04). This data suggests that sCgA correlates well with 5-year survival of GEP NET patients, and that IR-A mRNA expression correlates well with tumor mass in GEP NET patients. PMID:24042314

  5. Expression of Ki-67 (MIB-1) and GLUT-1 proteins in non-advanced prostatic cancer. (United States)

    Luczynska, Elzbieta; Gasinska, Anna; Wilk, Waclaw


    The expression of Ki-67 (MIB-1) and glucose transporter-1 (GLUT-1) were evaluated in patients with clinically localized prostate cancer (PC) who had undergone radical prostatectomy with curative intent. 140 low advanced PC specimens were studied. Protein expression was assessed immunohistochemically on tumour sections and expressed as a labelling index, i.e. the percentage of positively stained cells. In the case of Ki-67 nuclear staining and in the case of GLUT-1 membrane and cytoplasmic staining was considered as positive. The patients' mean age was 62.9 ±6.2 years. There were 13 (9.3%) at pTNM stage 1, 78 (55.7%) at stage 2, 40 (28.6%) at stage 3 and 9 (6.4%) at stage 4, respectively. 75 (53.6%) tumours were well differentiated (Gleason score ≤6), 52 (37.1%) moderately differentiated (Gleason score of 7) and 13 (9.3%) poorly differentiated (Gleason score 8-10). The mean pre-operative serum PSA was 9.9 ± SE 0.5 ng/ml, and the mean LI was equal to 8.1 ±0.6% and 29.7 ±2.0%, for MIB-1 and GLUT-1, respectively. Increase of pathological tumor volume and tumor grade was associated with statistically significant growth of PSA (p GLUT-1 LI the relation was not significant. Ki-67 expression was correlated with PSA levels (p = 0.013) and GLUT-1 scores (p = 0.04). In PC, an increase in the proliferation rate (higher MIB-1LI) in higher pTNM stages and tumour grades may point to Ki-67 as a good marker of biological aggressiveness useful in selecting patients for more aggressive treatment. A correlation between proliferation and GLUT-1 score may be the evidence of active glycolytic metabolism in hypoxic regions.


    Directory of Open Access Journals (Sweden)

    Luis Santamaría


    Full Text Available Cell proliferation, caspase 3 and pi-form of glutathione S transferase (GSTP1 were evaluated in prostate carcinoma (PCA, proliferative inflammatory atrophy (PIA and prostate intraepithelial neoplasia (PIN. Forty biopsies were classified as: without morphological lesions (controls: CTR, PIA, PIN and PCA. Ki67, caspase3 and GSTP1 were immunostained. The following estimates were performed: Numerical densities of Ki67+ cells (NVEPKi67, of all epithelial cells (NVEPtotal and of GSTP1+ cells (NVEPGSTP1; labelling index for Ki67 (LIKi67; volume fraction to caspase 3 positive tissue (VVcaspase 3 and of GSTP1 positive tissue (VVGSTP1. ANOVA was performed to compare the groups. NVEPtotal and NVEPKi67 were increased in PIA. LIKi67 was only increased in PCA. VVcaspase 3 was decreased in PIN and PCA. VVEGSTP1 was decreased in PCA. In our results PIA lacks the characteristics of a premalignant lesion. The result may be explained by the use of unbiased quantitative methods, the inadequate definition of PIA and the scarce inflammation observed in the samples with PIA included in this study.

  7. Expression of Pin1 and Ki67 in Cervical Cancer and Their Significance

    Institute of Scientific and Technical Information of China (English)


    In order to investigate the expression levels of Pin1 mRNA and protein in cervical cancer and its association with Ki67 and their clinical significance, amplification of Pin1 gene was examined by RT-PCR, and the expression of both Pin1 and Ki67 protein was detected by immunohistochemistry in cervical cancer tissues. It was shown that the expression levels of Pin1 were higher in cervical cancer than in normal cervical tissues (P<0.05). The expression of Pin1 protein was increased progressively along with the disease process from normal cervix to CIN and to cervical cancer (P<0. 05). No significant difference in the Pin1 expression was found between disease stages (FIGO),pathological grades or pelvic lymph node metastasis status (P>0.05). The expression of Pin1 was significantly higher in adenocarcinoma than insquamous carcinoma of the uterine cervix (P<0.05).In cervical cancer, the overexpression of Pin1 was positively correlated with that of Ki67 (P<0.05). These results suggested that the overexpression of Pin1 was closely related with cancer cell proliferation or progression of cervical cancer and contributed to oncogenesis. Pin1 may serve as a potential marker for cervical cancer diagnosis.

  8. Ki-67对早期乳腺癌治疗和预后的指导意义%Indicating significance of Ki-67 in therapy and prognosis of breast cancer

    Institute of Scientific and Technical Information of China (English)



    Ki-67,as one kind of nucleoantigen expressed in proliferated cells, has positive correlation with clinical stage, nodal status and Her-2,while negative correlation with estrogen receptor(ER). High Ki-67 expression influnced by endocrine therapy and chemotherapy indicates good proliferative activity. Ki-67, also as an independent prognostic factor of early breast cancer, provides prognostic information combined with other biologic markers. Ki-67 gives additional information breast cancer prognosis staging.%Ki-67是一种在增殖细胞中表达的核抗原,与临床分期、淋巴结状态和人表皮生长因子受体-2(Her-2)呈正相关,而与雌激素受体(ER)呈负相关.Ki-67高表达是细胞具有极度增殖活性的表现,内分泌治疗和化疗均对其表达有影响.Ki-67是早期乳腺癌的独立预后因素,亦可联合其他基因共同指导乳腺癌的预后,并可对乳腺癌的预后分级提供辅助信息.

  9. A Strategy for Screening Monoclonal Antibodies for Arabidopsis Flowers (United States)

    Shi, Qian; Zhou, Lian; Wang, Yingxiang; Ma, Hong


    The flower is one of the most complex structures of angiosperms and is essential for sexual reproduction. Current studies using molecular genetic tools have made great advances in understanding flower development. Due to the lack of available antibodies, studies investigating the localization of proteins required for flower development have been restricted to use commercial antibodies against known antigens such as GFP, YFP, and FLAG. Thus, knowledge about cellular structures in the floral organs is limited due to the scarcity of antibodies that can label cellular components. To generate monoclonal antibodies that can facilitate molecular studies of the flower, we constructed a library of monoclonal antibodies against antigenic proteins from Arabidopsis inflorescences and identified 61 monoclonal antibodies. Twenty-four of these monoclonal antibodies displayed a unique band in a western blot assay in at least one of the examined tissues. Distinct cellular distribution patterns of epitopes were detected by these 24 antibodies by immunofluorescence microscopy in a flower section. Subsequently, a combination of immunoprecipitation and mass spectrometry analysis identified potential targets for three of these antibodies. These results provide evidence for the generation of an antibody library using the total plant proteins as antigens. Using this method, the present study identified 61 monoclonal antibodies and 24 of them were efficiently detecting epitopes in both western blot experiments and immunofluorescence microscopy. These antibodies can be applied as informative cellular markers to study the biological mechanisms underlying floral development in plants. PMID:28293248

  10. Treatment with anti-interferon-δ monoclonal antibodies modifies experimental autoimmune encephalomyelitis in interferon-δ receptor knockout mice

    DEFF Research Database (Denmark)

    Espejo, C.; Penkowa, Milena; Saez-Torres, I.


    Neuroinflammation, neuronal degeneration, regeneration, monoclonal antibodies, multiple schlerosis......Neuroinflammation, neuronal degeneration, regeneration, monoclonal antibodies, multiple schlerosis...

  11. Drug Development of Therapeutic Monoclonal Antibodies. (United States)

    Mould, Diane R; Meibohm, Bernd


    Monoclonal antibodies (MAbs) have become a substantial part of many pharmaceutical company portfolios. However, the development process of MAbs for clinical use is quite different than for small-molecule drugs. MAb development programs require careful interdisciplinary evaluations to ensure the pharmacology of both the MAb and the target antigen are well-understood. Selection of appropriate preclinical species must be carefully considered and the potential development of anti-drug antibodies (ADA) during these early studies can limit the value and complicate the performance and possible duration of preclinical studies. In human studies, many of the typical pharmacology studies such as renal or hepatic impairment evaluations may not be needed but the pharmacokinetics and pharmacodynamics of these agents is complex, often necessitating more comprehensive evaluation of clinical data and more complex bioanalytical assays than might be used for small molecules. This paper outlines concerns and strategies for development of MAbs from the early in vitro assessments needed through preclinical and clinical development. This review focuses on how to develop, submit, and comply with regulatory requirements for MAb therapeutics.

  12. Expression and cinical significance of DLC-1 and Ki-67 proteins in gastric carcinoma%胃癌组织中DLC-1Ki-67蛋白的表达及临床意义

    Institute of Scientific and Technical Information of China (English)

    汤华; 赵铁


    Objective To investigate the expression of DLC - 1 and Ki - 67 in gastric cancer and adjacent normal gastric tissues and analyze the expression and its relation with clinicopathologic characteristics of gastric cancer and the prognosis. Methods The expression of DLC - 1 and Ki - 67 in 66 cases gastric cancer and 20 adjacent normal tissues was detected by immunohistochemiscal EnVision method and the relation of its expression with clinicopathologic characteristics of gastric cancer and the prognosis was analyzed. Results DLC - 1 and Ki - 67 were related to the differentiation, lymph node metastasis and clinical stages; no relation with gender and tumor location was found. DLC - 1 protein expression was negatively correlated with Ki - 67 protein expression ( rs = - 0. 67 ,P <0. 05 ). The 5 - year survival rate of DLC - 1 low expression and Ki - 67 high expression was significantly low. Conclusion Combining detection of DLC - 1 and Ki - 67 may be useful parameter for evaluating the biological behaviors of gastric cancer%目的 检测胃癌组织中DLC-1、Ki-67蛋白的表达,分析其与胃癌临床病理特征关系及预后的关系.方法 免疫组织化学EnVision两步法,检测66例胃癌和20例癌旁组织中DLC-1和Ki-67的表达,分析其表达与胃癌临床病理特征及预后的关系.结果 DLC-1 和Ki-67蛋白表达与肿瘤分化程度、淋巴结转移及TNM分期相关;与性别、肿瘤部位等无关;胃癌中DLC-1与Ki-67的表达呈负相关(rs=-0.673,P<0.05),且DLC-1低表达组及Ki-67高表达组术后5年生存率显著降低.结论 DLC-1和Ki-67有望成为胃癌生物学行为的参考指标.

  13. 18F-FDG PET/CT and extragastric MALT lymphoma: role of Ki-67 score and plasmacytic differentiation. (United States)

    Albano, Domenico; Bosio, Giovanni; Giubbini, Raffaele; Bertagna, Francesco


    The detection rate of fluorine-18-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) in extragastric mucosa associated lymphoid tissue (MALT) lymphoma is under debate and the reason is not clear. Our aim was to investigate the metabolic behavior of extragastric MALT lymphoma and whether the histological features (Ki-67 index and plasmacytic differentiation, PD) might explain it. PET/CT images were analyzed visually and semi-quantitatively and compared with Ki-67 index and PD. Seventy-two patients were included. Twelve of 18 patients with PD showed intense 18F-FDG uptake; on the contrary, 42 of 54 patients without PD had positive 18F-FDG PET/CT. Twenty-six of 27 patients with Ki-67 > 15% had 18F-FDG-avid lesions; 28 of 45 patients with Ki-67 ≤ 15% had positive 18F-FDG PET/CT. 18F-FDG avidity was significantly associated with Ki-67 index (p < .001) and not correlated with PD (p = .352). Maximum standardized uptake value (SUVmax), lesion-to-liver SUVmax ratio and lesion-to-blood pool SUVmax ratio were not correlated with Ki-67 index or PD. 18F-FDG avidity was noted in 75% and is correlated only with Ki-67.


    Institute of Scientific and Technical Information of China (English)

    SUN Da-wei; WANG Yan-jun; KONG Wei-jia; LIU Bang-hua


    Objective: To study the correlation between Ki67 and vascular endothelial growth factor(VEGF) and the significance of their expression in laryngeal squamous cell carcinoma(LSCC). Methods: The expressions of Ki67 and VEGF in 40 cases of LSCC and 5 cases of normal laryngeal mucosa were examined by immunohistochemical staining. Results: The expression levels of Ki67 and VEGF in LSCC tissue were higher than in normal laryngeal mucosa (Ki67: P<0.001, VEGF: P<0.001).The two indexes' levels in patients of different age or different sex had no significant difference (P>0.05). They were higher in LSCC with metastasis of lymph nodes than in patients without metastasis (Ki67: P=0.034, VEGF: P=0.006). The expressions of the two genes elevated correspondingly along with the development of LSCC T stage (P<0.05). In addition,correlation analysis indicated that the expression of Ki67 had a positive correlation with VEGF in LSCC(r=0.823, P<0.01).Conclusion: Ki67 and VEGF are objective indexes for the biological behavior of LSCC, and they might be helpful to the diagnosis, treatment and prognosis of laryngocarcinoma.

  15. Monoclonal antibodies against naturally occurring bioactive compounds. (United States)

    Shoyama, Y; Tanaka, H; Fukuda, N


    The ratio of hapten to bovine serum albumin (BSA) in an antigen conjugate was determined by matrix-assisted laser desorption/ionization (MALDI) tof mass spectrometry. A hybridoma secreting monoclonal antibody (MAb) was produced by fusing splenocytes immunized with an antigen-BSA conjugate with HAT-sensitive mouse myeloma cells. The cross-reaction of anti-forskolin antibodies with 7-deacetyl forskolin was 5.6%. A very small cross-reaction appeared with other derivatives. The full measuring range of the assay extends from 5 ng to 5 mug/ml of forskolin. Immunoaffinity column chromatography using anti-forskolin MAbs appears to be far superior to previously published separation methods. The capacity of the immunoaffinity column as determined by ELISA is 9 mug/ml. Forskolin has been isolated directly from the crude extracts of tuberous roots and the callus culture of Coleus forskohlii. A MAb against tetrahydrocannabinolic acid (THCA) was produced. The cross-reaction of anti-THCA antibody against other cannabinoids was very wide. Many cannabinoids and a spiro-compound were reactive, but did not react with other phenolics. It became evident that this ELISA was able to be applied to the biotransformation experiments of cannabinoids in plant tissue culture system. Anti-ginsenoside Rb1 MAbs were produced. New western blotting method of determination for ginsenosides was established. Ginsenosides separated by silica gel TLC were transferred to a polyvinylidene difluoride (PVDF) membrane. The membrane was treated with NaIO(4) solution followed by BSA, resulting in a ginsenoside-BSA conjugate. Immunostaining of ginsenosides was more sensitive compared to other staining. Immunostaining of ginsenosides in the fresh ginseng root was succeeded using anti-ginsenoside Rb1 (GRb1) MAb after blotting to PVDF membrane.

  16. Sub-Nanogram Detection of RDX Explosive by Monoclonal Antibodies. (United States)

    Ulaeto, David O; Hutchinson, Alistair P; Nicklin, Stephen


    Polyclonal and monoclonal antibodies were raised to protein carrier molecules haptenized with RDX, a major component of many plastic explosives including Semtex. Sera from immunized mice detected RDX protein conjugates in standard ELISA. Clonally purified monoclonal antibodies had detection limits in the sub-ng/mL range for underivatized RDX in competition ELISA. The monoclonal antibodies are not dependent on the presence of taggants added during the manufacturing process, and are likely to have utility in the detection of any explosive containing RDX, or RDX contamination of environmental sites.

  17. Monoclonal antibodies and Fc fragments for treating solid tumors

    Directory of Open Access Journals (Sweden)

    Eisenbeis AM


    Full Text Available Andrea M Eisenbeis, Stefan J GrauDepartment of Neurosurgery, University Hospital of Cologne, Cologne, GermanyAbstract: Advances in biotechnology, better understanding of pathophysiological processes, as well as the identification of an increasing number of molecular markers have facilitated the use of monoclonal antibodies and Fc fragments in various fields in medicine. In this context, a rapidly growing number of these substances have also emerged in the field of oncology. This review will summarize the currently approved monoclonal antibodies used for the treatment of solid tumors with a focus on their clinical application, biological background, and currently ongoing trials.Keywords: targeted therapy, monoclonal antibodies, cancer, biological therapy

  18. 直肠腺癌中血管内皮生长因子和细胞增生核抗原的表达及其与放化疗敏感性的关系%The predictive value of vascular endothelial growth factor and Ki-67 expression on neoadjuvant therapy in rectal cancer

    Institute of Scientific and Technical Information of China (English)

    Shumei Jiang; Renben Wang; Jinming Yu; Zhenjiang Zhang; Dianbin Mu; Zhongfa Xu


    Objective: To investigate the expressions of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (Ki-67) in patients with rectal adenocarcinoma and their associations with neoadjuvant therapy. Methods: The expressions of Ki-67 and VEGF in 32 cases of rectal adenocarcinoma, including both pretreatment tumor biopsies and postoperative specimen, were detected by immunohistochemistry using specific antibodies, and were correlated with clinico-pathological factors. Results: The intensity of VEGF staining was significantly correlated with lymph nodal metastasis (P = 0.033), depth of tumor invasion (P = 0.007) and tumor stage (P = 0.016), but not with histological types, tumor sizes, patients' ages and genders (P > 0.05). Low level of VEGF expression had significant correlation with the high sensitivity of response to neoadjuvant therapy (P = 0.016). The transient increase of VEGF expression could be seen after neoadjuvant therapy (P = 0.035). Ki-67 labeling index (Ki-67-LI) was significantly correlated with lymph node metastasis (P = 0.028), but not correlated to tumor sizes, patients' ages and genders (P > 0.05). Tumors with lower Ki-67-LI were more sensitive to neoadjuvant therapy (P = 0.032). The Ki-67 level decreased after neoadjuvant therapy, but no statistical significance was found (P > 0.05). Conclu-sion: Our results demonstrate that the expression of VEGF and Ki-67 in pretreatment rectal adenocarcinoma biopsies may be predictive of tumor response to neoadjuvant therapy.

  19. Production and characterization of monoclonal antibodies against mink leukocytes

    DEFF Research Database (Denmark)

    Chen, W.S.; Pedersen, Mikael; Gram-Nielsen, S.


    Three monoclonal antibodies (mAbs) were generated against mink leukocytes. One antibody reacted with all T lymphocytes, one with all monocytes and one had platelet reactivity. Under reducing conditions, the T lymphocyte reactive antibody immunoprecipitated 18 kDa, 23 kDa, 25 kDa and 32-40 kDa pol...

  20. Syngeneic anti-idiotypic monoclonal antibodies to an anti-NeuGc-containing ganglioside monoclonal antibody. (United States)

    Vázquez, A M; Pérez, A; Hernández, A M; Macías, A; Alfonso, M; Bombino, G; Pérez, R


    An IgM monoclonal antibody (MAb), named P3, has the characteristic to react specifically with a broad battery of N-glycolyl containing-gangliosides and with antigens expressed on breast tumors. When this MAb was administered alone in syngeneic mice, an specific IgG anti-idiotypic antibody (Ab2) response was induced, this Ab2 response was increased when P3 MAb was injected coupled to a carrier protein and in the presence of Freund's adjuvant. Spleen cells from these mice were used in somatic-cell hybridization experiments, using the murine myeloma cell line P3-X63-Ag8.653 as fusion partner. Five Ab2 MAbs specific to P3 MAb were selected. These IgG1 Ab2 MAbs were able to block the binding of P3 MAb to GM3(NeuGc) ganglioside and to a human breast carcinoma cell line. Cross-blocking experiments demonstrated that these Ab2 MAbs are recognizing the same or very close sites on the Abl MAb. The five Ab2 MAbs were injected into syngeneic mice and four of them produced strong anti-anti-idiotypic antibody (Ab3) response. While these Ab2 MAbs were unable to generate Ab3 antibodies with the same antigenic specificity than P3 MAb, three of them induced antibodies bearing P3 MAb idiotopes (Ag-Id+ Ab3). These results demonstrated that these Ab2 MAbs are not "internal image" antibodies, but they could define "regulatory idiotopes."

  1. MGMT和Ki-67抗原在肝细胞癌组织中的表达及意义%The Expressions and Clinical Significances of MGMT and Ki-67 Antigen in Human Hepatocellular Carcinoma

    Institute of Scientific and Technical Information of China (English)

    宋德霸; 赵龙栓; 王万鹏; 朱志杰; 黄修明; 谷渊


    目的 探讨O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)和细胞增殖核抗原Ki-67在原发性肝细胞肝癌(HCC)组织中的表达及意义.方法 应用免疫组化S-P法检测68例HCC组织及癌旁组织中MGMT及Ki-67抗原的表达.结果 MGMT、Ki-67抗原在HCc组织中的表达率分别为44.12%(30/68)和57.35%(39/68),MGMT的表达与Hcc组织病理分级有关(P0.05);Ki-67抗原的表达与HCC的肿瘤大小、包膜是否完整、脉管有无癌栓、病理分级有关(P0.05);并且随Ki-67抗原表达强度的增加,MGMT的表达强度明显下降,Ki-67抗原与MGMT表达呈负相关(γ=-0.301,P<0.05).结论 MGMT在HCC的发生发展中可能起抑制作用,MGMT的缺失是HCC发生的重要分子事件之一,检测Ki-67抗原有助于判断HCC恶性程度及复发转移的风险,联合检测MGMT和Ki-67抗原有助于预测HCC患者预后.

  2. Hot spot detection for breast cancer in Ki-67 stained slides: image dependent filtering approach (United States)

    Niazi, M. Khalid Khan; Downs-Kelly, Erinn; Gurcan, Metin N.


    We present a new method to detect hot spots from breast cancer slides stained for Ki67 expression. It is common practice to use centroid of a nucleus as a surrogate representation of a cell. This often requires the detection of individual nuclei. Once all the nuclei are detected, the hot spots are detected by clustering the centroids. For large size images, nuclei detection is computationally demanding. Instead of detecting the individual nuclei and treating hot spot detection as a clustering problem, we considered hot spot detection as an image filtering problem where positively stained pixels are used to detect hot spots in breast cancer images. The method first segments the Ki-67 positive pixels using the visually meaningful segmentation (VMS) method that we developed earlier. Then, it automatically generates an image dependent filter to generate a density map from the segmented image. The smoothness of the density image simplifies the detection of local maxima. The number of local maxima directly corresponds to the number of hot spots in the breast cancer image. The method was tested on 23 different regions of interest images extracted from 10 different breast cancer slides stained with Ki67. To determine the intra-reader variability, each image was annotated twice for hot spots by a boardcertified pathologist with a two-week interval in between her two readings. A computer-generated hot spot region was considered a true-positive if it agrees with either one of the two annotation sets provided by the pathologist. While the intra-reader variability was 57%, our proposed method can correctly detect hot spots with 81% precision.

  3. A monoclonal antibody against human MUDENG protein. (United States)

    Wagley, Yadav; Choi, Jun-Ha; Wickramanayake, Dimuthu Dhammika; Choi, Geun-Yeol; Kim, Chang-Kyu; Kim, Tae-Hyoung; Oh, Jae-Wook


    MUDENG (mu-2-related death-inducing gene, MuD) encodes a predicted ∼54-kDa protein in humans, considered to be involved in trafficking proteins from endosomes toward other membranous compartments as well as in inducing cell death. Here we report on the generation of a mouse monoclonal antibody (MAb) against the middle domain of human (h) MuD. This IgG sub 1 MAb, named M3H9, recognizes residues 244-326 in the middle domain of the MuD protein. Thus, the MuD proteins expressed in an astroglioma cell line and primary astrocytes can be detected by the M3H9 MAb. We showed that M3H9 MAb can be useful in enzyme-linked immunosorbent assay (ELISA) and immunoblot experiments. In addition, M3H9 MAb can detect the expression of the MuD protein in formalin-fixed, paraffin-embedded mouse ovary and uterus tissues. These results indicate that the MuD MAb M3H9 could be useful as a new biomarker of hereditary spastic paraplegia and other related diseases.

  4. Prognostic significance of MCM2, Ki-67 and gelsolin in non-small cell lung cancer

    Directory of Open Access Journals (Sweden)

    Huberman Joel


    Full Text Available Abstract Background Uncontrolled proliferation and increased motility are hallmarks of neoplastic cells, therefore markers of proliferation and motility may be valuable in assessing tumor progression and prognosis. MCM2 is a member of the minichromosome maintenance (MCM protein family. It plays critical roles in the initiation of DNA replication and in replication fork movement, and is intimately related to cell proliferation. Ki-67 is a proliferation antigen that is expressed during all but G0 phases of the cell cycle. Gelsolin is an actin-binding protein that regulates the integrity of the actin cytoskeletal structure and facilitates cell motility. In this study, we assessed the prognostic significance of MCM2 and Ki-67, two markers of proliferation, and gelsolin, a marker of motility, in non-small cell lung cancer (NSCLC. Methods 128 patients with pathologically confirmed, resectable NSCLC (stage I-IIIA were included. Immunohistochemistry was utilized to measure the expressions of these markers in formalin-fixed, paraffin-embedded tumor tissues. Staining and scoring of MCM2, Ki-67 and gelsolin was independently performed. Analyses were performed to evaluate the prognostic significance of single expression of each marker, as well as the prognostic significance of composite expressions of MCM2 and gelsolin. Cox regression and Kaplan-Meier survival analysis were used for statistical analysis. Results Of the three markers, higher levels of gelsolin were significantly associated with an increased risk of death (adjusted RR = 1.89, 95% CI = 1.17–3.05, p = 0.01, and higher levels of MCM2 were associated with a non-significant increased risk of death (adjusted RR = 1.36, 95% CI = 0.84–2.20, p = 0.22. Combined, adjusted analyses revealed a significantly poor prognostic effect for higher expression of MCM2 and gelsolin compared to low expression of both biomarkers (RR = 2.32, 95% CI = 1.21–4.45, p = 0.01. Ki-67 did not display apparent prognostic

  5. Generation and characterization of monoclonal antibodies specific to Coenzyme A

    Directory of Open Access Journals (Sweden)

    Malanchuk O. M.


    Full Text Available Aim. Generation of monoclonal antibodies specific to Coenzyme A. Methods. Hybridoma technique. KLH carrier protein conjugated with CoA was used for immunization. Screening of positive clones was performed with BSA conjugated to CoA. Results. Monoclonal antibody that specifically recognizes CoA and CoA derivatives, but not its precursors ATP and cysteine has been generated. Conclusion. In this study, we describe for the first time the production and characterization of monoclonal antibodies against CoA. The monoclonal antibody 1F10 was shown to recognize specifically CoA in Western blotting, ELISA and immunoprecipitation. These properties make this antiboby a particularly valuable reagent for elucidating CoA function in health and disease.

  6. Screening for cervical cancer precursors with p16/Ki-67 dual-stained cytology: results of the PALMS study

    National Research Council Canada - National Science Library

    Ikenberg, Hans; Bergeron, Christine; Schmidt, Dietmar; Griesser, Henrik; Alameda, Francisco; Angeloni, Claudio; Bogers, Johannes; Dachez, Roger; Denton, Karin; Hariri, Jalil; Keller, Thomas; von Knebel Doeberitz, Magnus; Neumann, Heinrich H; Puig-Tintore, Luis M; Sideri, Mario; Rehm, Susanne; Ridder, Ruediger


    ...) for the detection of high-grade cervical intraepithelial neoplasia (CIN2+). We assessed whether p16/Ki-67 dual-stained cytology, a biomarker combination indicative of transforming HPV infections, can provide high sensitivity for CIN2...

  7. Ki-67 proliferation index but not mitotic thresholds integrates the molecular prognostic stratification of lower grade gliomas (United States)

    Duregon, Eleonora; Bertero, Luca; Pittaro, Alessandra; Soffietti, Riccardo; Rudà, Roberta; Trevisan, Morena; Papotti, Mauro; Ventura, Laura; Senetta, Rebecca; Cassoni, Paola


    Despite several molecular signatures for “lower grade diffuse gliomas” (LGG) have been identified, WHO grade still remains a cornerstone of treatment guidelines. Mitotic count bears a crucial role in its definition, although limited by the poor reproducibility of standard Hematoxylin & Eosin (H&E) evaluation. Phospho-histone-H3 (PHH3) and Ki-67 have been proposed as alternative assays of cellular proliferation. Therefore in the present series of 141 LGG, the molecular characterization (namely IDH status, 1p/19q co-deletion and MGMT promoter methylation) was integrated with the tumor “proliferative trait” (conventional H&E or PHH3-guided mitotic count and Ki-67 index) in term of prognosis definition. Exclusively high PHH3 and Ki-67 values were predictor of poor prognosis (log rank test, P = 0.0281 for PHH3 and P = 0.032 for Ki-67), unlike standard mitotic count. Based on Cox proportional hazard regression analyses, among all clinical (age), pathological (PHH3 and Ki-67) and molecular variables (IDH, 1p/19q codeletion and MGMT methylation) with a prognostic relevance at univariate survival analysis, only IDH expression (P = 0.001) and Ki-67 proliferation index (P = 0.027) proved to be independent prognostic factors. In addition, stratifying by IDH expression status, high Ki-67 retained its prognostic relevance uniquely in the IDH negative patient (P = 0.029) doubling their risk of death (hazard ratio = 2.27). Overall, PHH3 immunostaining is the sole reliable method with a prognostic value to highlight mitotic figures in LGG. Ki-67 proliferation index exceeds PHH3 mitotic count as a predictor of patient's prognosis, and should be integrated with molecular markers in a comprehensive grading system for LGG. PMID:27049832

  8. Purification of Murine Monoclonal IgM Antibody

    Institute of Scientific and Technical Information of China (English)


    This paper presents the purification of a monoclonal IgM antibody against human tumor associated antigen Lewis-Y by ion exchange chromatography and gel filtration.Enzyme-linked immunosorbent assay (ELISA) and SDS-polyacrylamide gel electrophoresis (PAGE) were used to identify purified IgM antibody.In flow cytometry analysis, the purified IgM antibody recognizes human breast tumor cell line MCF-7 which expresses Lewis-Y antigen.This work presents a new way for the purification of murine monoclonal IgM antibody.

  9. Relationship of doxorubicin- and radiation-induced apoptosis with Ki-67 labeling index in human tumors in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Hayakawa, Kayoko; Hasegawa, Masatoshi; Kawashima, Miho; Toda, Hisako; Hayakawa, Kazushige; Mitsuhashi, Norio; Niibe, Hideo [Gunma Univ., Maebashi (Japan). School of Medicine


    In the use of doxorubicin and radiation for treatment of human malignant tumors in vivo, the relationship between treatment-induced apoptosis and Ki-67 labeling index was investigated. Four human tumor xenografts (ependymoblastoma, NNE; primitive neuroectodermal tumor, YKP; small cell lung carcinoma, GLS; glioblastoma, KYG) were transplanted under the skin of thigh of the nude mice (BALB/cA JcL-nu). The mice were given a single radiation dose of 1 Gy, or doxorubicin alone intraperitoneally at a dose of 8 mg/kg. After treatment, sections of tumor specimens were prepared from paraffin-embedded tissues. Hematoxylin and eosin staining, TUNEL staining, and immunohistochemical analysis of Ki-67 were performed. In NNE, apoptotic cells appeared most frequently after treatment compared with all other tumors, and the incidence of apoptosis in the radiation-treated group was much higher than in the doxorubicintreated group. As the incidence of apoptosis in NNE increased, the Ki-67 labeling index tended to decrease. In GLS and KYG, there was a low incidence of treatment-induced apoptosis, although the Ki-67 labeling index decreased transiently after treatment. In YKP, few apoptotic cells appeared and Ki-67 the labeling index was unchanged throughout the time course after treatment. Ki-67 labeling index in malignant tumors after treatment may be affected by various kinds of cell deaths and treatment methods. (author)

  10. Suprabasal expression of Ki-67 as a marker for the severity of oral epithelial dysplasia and oral squamous cell carcinoma

    Directory of Open Access Journals (Sweden)

    Nidhi Dwivedi


    Full Text Available Background: Transition of the normal oral epithelium to dysplasia and to malignancy is featured by increased cell proliferation. To evaluate the hypothesis of distributional disturbances in proliferating and stem cells in oral epithelial dysplasia and oral squamous cell carcinoma (OSCC. Aim: To evaluate layer wise expression of Ki-67 in oral epithelial dysplasia and in OSCC. Materials and Methods: Thirty histologically confirmed cases of oral epithelial dysplasia, fifteen cases of OSCC and five cases of normal buccal mucosa were immunohistochemically examined and nuclear expression of Ki-67 was counted according to basal, parabasal, and suprabasal layers in epithelial dysplasia and number of positive cells per 100 cells in OSCC as labeling index (LI. Results: Suprabasal expression of Ki-67 increased according to the severity of epithelial dysplasia and the difference was statistically significant ( P < 0.001. The mean Ki-67LI was 12.78 for low risk lesions, 28.68 for high risk lesions, 39.45 for OSCC and 13.6 for normal buccal mucosa. Conclusion: The results of the present study demonstrate the use of proliferative marker Ki-67 in assessing the severity of epithelial dysplasia. Suprabasal expression of Ki-67 provides an objective criteria for determining the severity of epithelial dysplasia and histological grading of OSCC.

  11. Characterization of human papillomavirus infection, P53 and Ki-67 expression in cervix cancer of Mozambican women. (United States)

    Carrilho, Carla; Gouveia, Patricia; Cantel, Martha; Alberto, Matos; Buane, Landim; David, Leonor


    In this study, we aimed at evaluating the distribution of HPV types and the expression of P53 and Ki-67 in cervix carcinomas of Mozambican women. Fourty-seven invasive carcinomas, 10 CIN III, and 10 normal cervix were studied. P53 and Ki-67 expression was examined immunohistochemically. HPV infection and HPV types were detected by PCR (GP5+/bio-GP6+) and enzyme-immunoassay, respectively. Expression of P53 and Ki-67 and detection of HPV were as follows: normal cervix--0%, 10%, and 0%, respectively; CIN III--10%, 0%, and 100%, respectively; invasive carcinomas--50%, 55.5%, and 70%, respectively. HPV 16 was identified in 54% of invasive carcinomas, HPV 31, 33, 35, and 45 in 23%, "unidentified" HPV in 19%, and HPV 18 in 4% of invasive carcinomas. No significant associations were observed between P53 expression, Ki-67 expression, and HPV infection. In conclusion, we observed a high frequency of HPV infection in CIN III lesions and invasive carcinomas from Mozambican women, with HPV 16 representing the most frequent viral type. HPV status was not related to P53 and Ki-67 expression. Both P53 and Ki-67 are associated with invasive cervix carcinomas, mainly of the squamous keratinizing histotype.

  12. Quantum dots-based double imaging combined with organic dye imaging to establish an automatic computerized method for cancer Ki67 measurement (United States)

    Wang, Lin-Wei; Qu, Ai-Ping; Liu, Wen-Lou; Chen, Jia-Mei; Yuan, Jing-Ping; Wu, Han; Li, Yan; Liu, Juan


    As a widely used proliferative marker, Ki67 has important impacts on cancer prognosis, especially for breast cancer (BC). However, variations in analytical practice make it difficult for pathologists to manually measure Ki67 index. This study is to establish quantum dots (QDs)-based double imaging of nuclear Ki67 as red signal by QDs-655, cytoplasmic cytokeratin (CK) as yellow signal by QDs-585, and organic dye imaging of cell nucleus as blue signal by 4‧,6-diamidino-2-phenylindole (DAPI), and to develop a computer-aided automatic method for Ki67 index measurement. The newly developed automatic computerized Ki67 measurement could efficiently recognize and count Ki67-positive cancer cell nuclei with red signals and cancer cell nuclei with blue signals within cancer cell cytoplasmic with yellow signals. Comparisons of computerized Ki67 index, visual Ki67 index, and marked Ki67 index for 30 patients of 90 images with Ki67 ≤ 10% (low grade), 10% dye imaging on BC tissues, this study successfully developed an automatic computerized Ki67 counting method to measure Ki67 index.

  13. Pokemon、Ki-67及Livin蛋白在胃癌中的表达及意义%Expression and clinical signofocance of Pokemon, Ki-67 and Livin protein in gastric cancer

    Institute of Scientific and Technical Information of China (English)

    李月锋; 曲宏岩; 张东欣


    Objective: To study the expression of Pokemon、Ki-67、Livin and its clinical significant in gastric cancer and normal of gastric mucosa. Methods: The Pokemon, Ki-67 and Livin in gastric cancer(n=80), normal gastric mucosa(n=80) was detected by Western-blot. Results: Pokemon protein expression was high in of cases in gastric cancer, which was significantly different from the expression in normal gastric mucosa, expression of Pokemon was relative to high and low of hyperplasia of gastric mucosa. It was highly expressed in gastric cancer and was related with stage of TNM and lymphoid node transfer. But were not related with the age,sex and the size of tumor. Conclusion: Pokemon, Ki-67 and Livin protein expression may be correlate with the carcinogenesis and genesis, development and metastasis of gastric cancer. Pokemon may be an effective target gene for therapy of gastric cancer, accordingly, which may develop a new way to gene therapy for gastric cancer.%目的:检测胃癌组织中Pokemon、Ki-67及Livin的表达,探讨其在胃癌的表达及意义.方法:应用Western-Blot方法检测80例胃癌标本及正常胃组织中Pokemon、Ki-67及Livin的表达.结果:胃癌组织中Pokemon、Ki-67及Livin的表达分别为67.5% (54/80)、42.5% (34/80)和52.5% (42/80).正常胃组织中Pokemon、Ki-67及Livin的表达为12.5% (10/80)、7.5%(6/80)、11.3% (9/80).胃癌组织中Pokemon、Ki-67、Livin的表达与临床TNM分期、淋巴结转移有关,与年龄、性别、部位、Borrman分型无关.Pokemon与Ki-67、Livin的表达呈正相关(P<0.05).结论:Pokemon、Ki-67、Livin共同参与了胃癌的发生、发展和转移,可能成为胃癌临床诊断指标.

  14. 探讨 APC、P53、Ki-67对大肠腺瘤诊断的意义%Investigation of significance of APC, P53, Ki-67 in diagnosis of colorectal adenoma

    Institute of Scientific and Technical Information of China (English)



    目的:探求结肠腺瘤样息肉蛋白(APC)、P53、Ki-67在大肠腺瘤的不同发展阶段中的表达,总结 APC、P53、Ki-67在大肠腺瘤诊断中的价值及分析其对临床的指导意义。方法大肠腺瘤患者132例,分别采用 SP 免疫组化染色,比较免疫组化结果。结果在大肠腺瘤伴高级别上皮内瘤变中P53强表达,而在大肠腺瘤伴低级别上皮内瘤变中 APC 强表达(P<0.05);在表达 Ki-67≥20%的患者中 P53强表达,而在 Ki-67<20%的患者中 APC 强表达(P<0.05)。结论APC、P53、Ki-67联合标记对大肠腺瘤的病理诊断起到辅助作用,指导临床对患者预后的判断,做进一步临床处理。%Objective To investigate expressions of adenomatosis polyposis coli (APC), P53, and Ki-67 in different stages of colorectal adenoma, to summarize value of APC, P53 and Ki-67 in diagnosis of colorectal adenoma, and to analyze their clinical guidance significance. Methods A total of 132 colorectal adenoma patients received SP immumohistochemical staining. Their immumohistochemical outcomes were compared. Results P53 showed strong expression in colorectal adenoma complicated with high grade intraepithelial neoplasia, while APC showed strong expression in colorectal adenoma complicated with low grade intraepithelial neoplasia (P<0.05). In patients with Ki-67 ≥ 20%, P53 had its strong expression, while APC showed strong expression in those with Ki-67 <20% (P<0.05). Conclusion Combined marker of APC, P53 and Ki-67 provides assisted effect in pathological diagnosis of colorectal adenoma. This method provides clinical guidance for prognosis judgment and further clinical management.

  15. Malignant adenohypophysis spindle cell oncocytoma with repeating recurrences and a high Ki-67 index (United States)

    Kong, Xiangyi; Li, Dongmei; Kong, Yanguo; Zhong, Dingrong


    Abstract Adenohypophysis spindle cell oncocytoma (ASCO) is a rare tumor recently reported by Roncaroli et al in 2002. This tumor is considered a grade I tumor by the World Health Organization. We report a rare case of malignant ASCO with repeating recurrences and a high Ki-67 index—a challenging diagnosis guided by clinical presentations, radiological signs, and postoperative pathological tests. We represent a 30-year-old man who had suffered from headaches, diplopia, and impaired visual field and acuity. His magnetic resonance imaging revealed an abnormal sellar mass and was originally misdiagnosed as a pituitary macroadenoma. We present detailed analysis of the patient's disease course and review relevant literature. When surgically treated, the specimen revealed a typical histopathology pattern of ASCO. The tumor recurred for several times and the patient underwent 3 surgeries and 1 γ-knife treatment, which was accompanied by a continuously increasing Ki-67 index. This is the first reported case of malignant ASCO (WHO III–IV grade). Despite its rarity, ASCO should be considered in the differential diagnosis of sellar lesions that mimic pituitary adenomas. PMID:28121922

  16. ELISA Detection of Francisella tularensis using Polyclonaland Monoclonal Antibodies

    Directory of Open Access Journals (Sweden)

    Miroslav Pohanka


    Full Text Available The mouse monoclonal and polyclonal antibodies were produced for the detection of intracellular pathogenand potential warfare agent Francisella tularensis. Antibody titers obtained were 1:640 for polyclonal antibodiesand 1:320 for monoclonal antibodies. Both antibodies were used in the indirect enzyme-linked immunosorbentassay (ELISA found to detect F. tularensis whole cells. The limit of detection was 5.4×106 CFU/ml for polyclonalantibodies and 6.9×106 CFU/ml for monoclonal antibodies. The value sample could  be distinguished from anyconcentration of another gram-negative bacterium: Escherichia coli.Defence Science Journal, 2008, 58(5, pp.698-702, DOI:

  17. MDM-2、bcl-2 和Ki-67在子宫颈癌中的表达及其意义%Expression and Evaluation of MDM-2, bcl-2 and Ki-67 in Cervical Carcinoma

    Institute of Scientific and Technical Information of China (English)

    李宏艳; 张建中


    目的 探讨MDM-2、bcl-2和Ki-67在子宫颈癌的发生、发展过程中的作用.方法 采用免疫组化法检测MDM-2、bcl-2和Ki-67在31例子宫颈癌、13例子宫颈非典型性增生、15例正常子宫颈组织中的表达水平.结果 在31例子宫颈癌组织中MDM-2表达阳性11例,bcl-2阳性13例,Ki-67阳性15例.bcl-2、MDM-2和Ki-67三者共同阳性9例(29.03%).对31例子宫颈癌组织中MDM-2与bcl-2共同阳性者、MDM-2与Ki-67共同阳性者及Ki-67与bcl-2共同阳性者进行配对资料χ2检验,P<0.05.子宫颈癌组织中bcl-2、MDM-2和Ki-6阳性表达率彼此间存在着明显的正性相关关系.结论 联合检测MDM-2、bcl-2和Ki-67在子宫颈癌中的表达,显示三者具有明显相关性,与子宫颈癌的发生、发展有关.

  18. Survivin、Ki67在牙源性囊肿上皮组织中的表达及意义%The expression and significance of Survivin and Ki67 in odontogenic jaw cyst epithelium

    Institute of Scientific and Technical Information of China (English)

    刘喜玲; 陶学金; 袁健; 汤国雄; 马净植



  19. Development of monoclonal antibodies that recognize Treponema pallidum.


    Saunders, J M; Folds, J D


    We developed a panel of monoclonal antibodies to Treponema pallidum (Nichols) antigens, some of which recognize treponemal antigens on T. pallidum (Nichols), T. pallidum strain 14, and Treponema phagedenis biotype Reiter. The antibodies were detected by either an enzyme-linked immunosorbent assay or a radioimmunoassay.

  20. Monoclonal antibodies for the control of influenza virus vaccines.

    NARCIS (Netherlands)

    H.J.M. van de Donk; M.F. van Olderen; A.D.M.E. Osterhaus (Albert); J.C. de Jong (Jan)


    textabstractHybridomas producing haemagglutination inhibiting monoclonal antibodies against influenza A/Texas/1/77 H3N2 were developed. One hybridoma producing antibodies reacting with Victoria/3/75, Texas/1/77 Bangkok/1/79 and England/496/80 was selected to determine the potency of influenza virusv


    Directory of Open Access Journals (Sweden)

    Duarte Keila M.R.


    Full Text Available Monoclonal antibodies were obtained against Tomato mosaic tobamovirus (ToMV isolated in Brazil. One antibody (8G7G2 isotyped as IgG2b (kappa light chain showed strong specificity and very low cross reaction with the Tobacco mosaic virus (TMV. It can be used in identification of tomato mosaic virus (ToMV.

  2. Serological comparison of tospovirus isolates using polyclonal and monoclonal antibodies.

    NARCIS (Netherlands)

    Adam, G.; Peters, D.; Goldbach, R.W.


    A test was conducted to compare tospovirus isolates using different poly- and monoclonal antibodies. All isolates and antibodies were compared under identical conditions. From 130 tospovirus isolates, which were obtained from all over the world and included well-characterized isolates from all four

  3. Serological comparison of tospovirus isolates using polyclonal and monoclonal antibodies.

    NARCIS (Netherlands)

    Adam, G.; Peters, D.; Goldbach, R.W.


    A test was conducted to compare tospovirus isolates using different poly- and monoclonal antibodies. All isolates and antibodies were compared under identical conditions. From 130 tospovirus isolates, which were obtained from all over the world and included well-characterized isolates from all four

  4. A new monoclonal antibody against DNA ligase I is a suitable marker of cell proliferation in cultured cell and tissue section samples

    Directory of Open Access Journals (Sweden)

    B Vitolo


    Full Text Available The extensive characterization of the replicative human DNA ligase I (LigI undertaken in the last decade demonstrated that the level of this protein strongly correlates with the rate of cell proliferation. This may allow to expand the repertoire of clinical biomarkers for the analysis of cell proliferation.We have produced a new monoclonal antibody (5H5 against LigI and exploited it as cell proliferation marker in Western blotting and immunofluorescence as well as in immunohistochemistry on paraffin tissue sections. The Western blot analysis showed that the LigI level detected by 5H5 antibody is high in all proliferating cells. On the contrary the protein is down regulated in resting human fibroblast and peripheral blood lymphocytes. Immunofluorescence analysis on cultured HeLa cells showed that 5H5 antibody labels all proliferating cells and displays the same staining pattern of BrdU in S-phase nuclei. Finally the analysis of serial sections of inflamed tonsils and NHL lymph nodes (either frozen or paraffin embedded demonstrated that 5H5 marks the same population of cells as the Ki-67 antibody. Our results demonstrate that 5H5 antibody is a valuable tool for labeling proliferating cells that can be conveniently used in Western blotting, immunocytochemistry and immunohistochemistry.

  5. Clinical significance of Ki-67, Kras and Braf protein expressions in gliomas%脑胶质瘤中Ki-67、Kras和Braf蛋白的表达及临床意义

    Institute of Scientific and Technical Information of China (English)

    姜振富; 李梅; 韦鸿; 王朝晖; 魏明海


    Objective:To study the clinical significance of Ki -67,Kras and Braf protein expressions in gliomas. Methods: Immunohistochemistry method ( SP) was performed to detect the expressions of Ki - 67, Kras and Braf protein in 81 gliomas from operation. We analyzed the correlations of the protein expressions to the clinical pathological a-gents by SPSS statistic software. Results:The positive expression of Ki -67 protein was 54. 3% in gliomas,and it were 0,37.0% ,65.4% and 73. 9% respectively in grade 1,2,3 and 4,there was significant difference in the four groups (P0.05). The expressions of Kras and Braf protein were 53. 1% and 61.7% .their expressions had no significant relation to the pathology grades,sex and age of patients (P>0. 05). The expression of Kras protein had no significant correlation to that of Braf (P >0.05). The expressions of Kras and Braf protein had no significant correlation to that of Ki - 67 (P > 0.05). Conclusion: The expression of Ki - 67 protein had no significant relation to the malignant degree of gliomas; Detection of Ki - 67 protein expression may be helpful for us to judge the malignant degree of gliomas ; In gliomas, the expressions of Kras and Braf protein were independent, it would suggest that there may have many mechanisms in the process of gliomas.%目的:了解Ki-67、Kras和Braf蛋白在脑胶质瘤中的表达状况,并探讨其临床意义.方法:免疫组织化学SP法检测81例手术切除脑胶质瘤组织的Ki-67、Kras和Braf蛋白表达情况.分析Kras、Braf和Ki-67蛋白的表达与临床病理学参数之间的关系.结果:胶质瘤中Ki-67蛋白表达阳性率为54.3%,在1、2、3、4级胶质瘤中分别为0、37.0%、65.4%、73.9%,各组间差异显著(P<0.05),但其表达与患者性别、年龄无明确相关关系(P >0.05);Kras蛋白表达阳性率为53.1%,Braf蛋白表达阳性率为61.7%,二者表达与肿瘤病理分级及患者性别、年龄无关(P>0.05).Kras与Braf蛋白表

  6. 宫颈鳞癌组织中Ki-67和P53的表达及其意义%Expressions and clinical significances of Ki-67 and P53 in cervical squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    齐斯琴; 德胜; 李冬梅; 石搏; 黄可欣


    目的:探讨宫颈鳞癌组织中Ki-67和P53蛋白的表达及其与临床病理特征的关系.方法:采用免疫组织化学技术,检测60例子宫颈鳞癌、40例宫颈原位癌(CINⅢ)和20例癌旁正常宫颈上皮(NCE)中Ki-67和P53的表达,分析其表达与宫颈鳞癌的临床分期、病理类型及有无淋巴结转移的关系.结果:Ki-67在宫颈鳞癌、原位癌和NCE中的阳性表达率分别为78.33%、52.50%、35.00%;P53为71.67%、55.00%、0.00%.NCE组与鳞癌组及原位癌组相比较差异均有统计学意义(P<0.01).Ki-67和P53的阳性表达率与临床分期、病理类型及有无淋巴结转移均有明显相关性(P<0.01);Ki-67和P53在宫颈鳞癌的表达中呈正相关(P<0.01).结论:Ki-67和P53在宫颈癌组织中表达水平的上调可能在宫颈鳞癌浸润和转移中起重要作用.%Objective: To explore the relationship between expressions of Ki -67 and P53 in cervical squamous cell carcinoma and the clinicopathological characteristics. Methods: Immunohistochemical method was used to detect the expression levels of Ki -67 and P53 in 60 cases with cervical squamous cell carcinoma, 40 cases with cervical carcinoma in situ ( CIN Ⅲ) and 20 cases with normal cervical epithelium adjacent to carcinoma (NCE) . The relationship between their expression levels and clinical stages, pathological types, and lymph node metastasis or not was analyzed. Results: The positive expression rates of Ki - 67 in cervical squamous cell carcinoma, cervical carcinoma in situ and NCE were 78. 33% , 52. 5% , and 35. 00% , respectively; the positive expression rates of P53 in cervical squamous cell carci- noma, cervical carcinoma in situ, and NCE were 71. 67% , 55. 00% , and 0.00% , respectively. There were statistically significant differences in positive expression rates of Ki - 67 and P53 between NCE group and cervical squamous cell carcinoma group, cervical carcinoma in situ group (P <0. 01) . There was a significant

  7. 视网膜母细胞瘤组织中 Ki-67、PTEN 表达的变化%Expressions of Ki-67 and PTEN in the Retinoblastoma

    Institute of Scientific and Technical Information of China (English)



    Objective To investigate the expressions of Ki-67 and PTEN in the retinoblastoma and the signifi-cance. Methods Immunohistochemical S-P method was used to detect the expressions of Ki-67 and PTEN in the 97 patients with retinoblastoma and 28 patients with normal retina tissues,their relationship with clinicopathological pa-rameters were analyzed. Results The positive rate of Ki-67 was 66. 0%(64 / 97)in the retinoblastoma,and was 14. 3%(4 / 28)in the normal retina tissues( χ2 = 23. 406,P < 0. 05). The positive rate of Ki-67 was 53. 6%(52 / 97)in the retinoblastoma,and was 92. 9%(26 / 28)in the normal retina tissues(χ2 = 14. 266,P < 0. 05). The expressions of Ki-67 and PTEN in the retinoblastoma were related with cell differentiation degree and clinical stage (P < 0. 05). In the retinoblastoma,the expression of Ki-67 was negatively related with PTEN(r = - 0. 493,P <0. 05). Conclusion Abnormal expressions of Ki-67 and PTEN may be related to the infiltration and development of retinoblastoma.%目的:探讨视网膜母细胞瘤组织中 Ki-67、PTEN 表达的变化及其临床意义。方法采用免疫组化S-P 法检测97例视网膜母细胞瘤和28例正常视网膜组织中 Ki-67、PTEN 的表达水平,并分析两者与视网膜母细胞瘤临床病理参数的关系。结果视网膜母细胞瘤组织中 Ki-67的阳性率为66.0%(64/97),高于正常视网膜组织的14.3%(4/28)(χ2=23.406,P <0.05);视网膜母细胞瘤组织中 PTEN 的阳性率为53.6%(52/97),低于正常视网膜组织的92.9%(26/28)(χ2=14.266,P <0.05)。视网膜母细胞瘤组织中 Ki-67、PTEN 的表达均与患者的分化程度、临床分期有关(P <0.05)。视网膜母细胞瘤组织中 Ki-67、PTEN 的表达呈负相关关系(r =-0.493,P <0.05)。结论 Ki-67、PTEN 的异常表达参与了视网膜母细胞瘤的侵袭和病程进展。

  8. High throughput production of mouse monoclonal antibodies using antigen microarrays

    DEFF Research Database (Denmark)

    De Masi, Federico; Chiarella, P.; Wilhelm, H.;


    Recent advances in proteomics research underscore the increasing need for high-affinity monoclonal antibodies, which are still generated with lengthy, low-throughput antibody production techniques. Here we present a semi-automated, high-throughput method of hybridoma generation and identification....... Monoclonal antibodies were raised to different targets in single batch runs of 6-10 wk using multiplexed immunisations, automated fusion and cell-culture, and a novel antigen-coated microarray-screening assay. In a large-scale experiment, where eight mice were immunized with ten antigens each, we generated...

  9. Interobserver reproducibility and accuracy of p16/Ki-67 dual-stain cytology in cervical cancer screening. (United States)

    Wentzensen, Nicolas; Fetterman, Barbara; Tokugawa, Diane; Schiffman, Mark; Castle, Philip E; Wood, Shannon N; Stiemerling, Eric; Poitras, Nancy; Lorey, Thomas; Kinney, Walter


    Dual-stain cytology for p16 and Ki-67 has been proposed as a biomarker in cervical cancer screening. The authors evaluated the reproducibility and accuracy of dual-stain cytology among 10 newly trained evaluators. In total, 480 p16/Ki-67-stained slides from human papillomavirus-positive women were evaluated in masked fashion by 10 evaluators. None of the evaluators had previous experience with p16 or p16/Ki-67 cytology. All participants underwent p16/Ki-67 training and subsequent proficiency testing. Reproducibility of dual-stain cytology was measured using the percentage agreement, individual and aggregate κ values, as well as McNemar statistics. Clinical performance for the detection of cervical intraepithelial neoplasia grade 2 or greater (CIN2+) was evaluated for each individual evaluator and for all evaluators combined compared with the reference evaluation by a cytotechnologist who had extensive experience with dual-stain cytology. The percentage agreement of individual evaluators with the reference evaluation ranged from 83% to 91%, and the κ values ranged from 0.65 to 0.81. The combined κ value was 0.71 for all evaluators and 0.73 for cytotechnologists. The average sensitivity and specificity for the detection of CIN2+ among novice evaluators was 82% and 64%, respectively; whereas the reference evaluation had 84% sensitivity and 63% specificity, respectively. Agreement on dual-stain positivity increased with greater numbers of p16/Ki-67-positive cells on the slides. Good to excellent reproducibility of p16/Ki-67 dual-stain cytology was observed with almost identical clinical performance of novice evaluators compared with reference evaluations. The current findings suggest that p16/Ki-67 dual-stain evaluation can be implemented in routine cytology practice with limited training. © 2014 American Cancer Society.

  10. The correlation between Ki-67 with other prognostic factors in breast cancer: A study in Iranian patients

    Directory of Open Access Journals (Sweden)

    Seyed-Hamid Madani


    Full Text Available Context: Despite the fact that breast cancer (BC is a major health issue, very few studies describe its characteristics in the Middle East. Aim: The aim of this study was to evaluate the use and value of Ki-67 as a prognostic marker in BC and associations between Ki-67, clinical, and histopathological parameters were evaluated. Subjects and Methods: In a retrospective study, 260 BC women and invasive ductal carcinoma were included to our study in Kermanshah city, Iran. Age, tumor size, lymph node involvement, histological grade, nuclear grade, and vascular invasion were other factors that determined in a lot of patients. Results: The mean age at diagnosis was 47.6 years (range, 24-84 years with 100% female. Of 243 patients that tumor size was determined for them, 207 patients (85.2% had tumor size ≥ 2 cm, and 36 patients (14.8% had size <2 cm and also of 237 patients, 47 patients (19.8%, 140 (59.1%, and 50 (21.1% had histological grades I, II, and III, respectively. There is significant correlation between Ki-67 with nuclear grade, human epidermal growth factor receptor 2 (HER2, and p53 (P < 0.05. Based on this result, more patients with Ki-67 ≥ 20% have higher nuclear grade, p53-positive, and HER2-positive. There was correlation between Ki-67 with type of tumor (P = 0.009. Conclusions: The higher Ki-67 has a direct significant correlation with higher nuclear grade, p53-positive, and HER2-positive. Furthermore, triple negative patients have higher Ki-67 compared to other subtypes.

  11. Prognostic value of Ki-67 in breast cancer patients with positive axillary lymph nodes: a retrospective cohort study.

    Directory of Open Access Journals (Sweden)

    Feng-yan Li

    Full Text Available INTRODUCTION: Ki-67 expression is a biomarker for proliferation. Its prognostic value is recognized in breast cancer (BC patients with negative axillary nodes, but is less clear in BC patients with positive axillary lymph nodes. METHODS: We retrospectively reviewed the medical records of 1131 Chinese BC patients treated from January 2002 to June 2007 and 450 patients met the inclusion criteria: positive nodes, adjuvant therapy, and complete biomarker profile (estrogen receptor (ER, progesterone receptor (PR, HER2, p53, Ki-67. Univariate and multivariate regression analysis were used to correlate biomarkers and tumor characteristics with metastasis free survival (MFS and overall survival (OS. RESULTS: Median follow-up time was 46 months (range 5-76 months. The Ki-67 expression was associated significantly with histological grade, ER, PR, HER2, and P53 status (P<0.05. Tumor stage, nodal stage, and ER status were independent prognostic factors for MFS. Ki-67 status was associated significantly with OS but not MFS. To determine whether the extent of LN involvement in the BC patients influenced the role of Ki-67 in survival rates, we compared these variables in patients with 1-3 positive lymph nodes (N1 to those of patients with ≥ 4 positive lymph nodes. Ki-67 status was an independent prognostic factor for MFS (Hazard Ratio, 3.27, P = 0.026 and overall survival (HR, 10.64, P = 0.007 in patients with 1-3 positive nodes (N1. CONCLUSIONS: The possibility that Ki-67 expression together with clinical factors can improve prediction of the prognosis of BC patients with 1 ∼ 3 positive axillary lymph nodes warrants further studies.

  12. Immunoblotting with monoclonal antibodies: importance of the blocking solution. (United States)

    Hauri, H P; Bucher, K


    Four commonly used blocking agents, i.e., fetal calf serum, mammalian gelatin-Nonidet-P40, fish gelatin-Nonidet-P40, and defatted powdered milk were compared with respect to their efficiency to block the nonspecific background and to promote maximal immunoreactivity of monoclonal antibodies against human intestinal sucrase-isomaltase during immunoblotting. Two of five monoclonal antibodies were found to react with the electroblotted enzyme. However, one of the reacting antibodies gave optimal results with fish gelatin-Nonidet-P40 and the other with defatted powdered milk, while fetal calf serum lead to unacceptably high backgrounds. The results suggest that some of the difficulties encountered with monoclonal antibodies in immunoblotting may be due to inappropriate blocking conditions.

  13. Expression of Ki-67/caspase-3 cocktail double staining in prostatic adenocarcinoma%前列腺腺癌中双染鸡尾酒抗体Ki-67/caspase-3的表达及意义

    Institute of Scientific and Technical Information of China (English)

    孙平丽; 金仁顺


    目的 探讨Ki-67/caspase-3鸡尾酒抗体在前列腺腺癌(PAC)、高级别前列腺上皮内瘤(HGPIN)及良性前列腺增生(BPH)组织中的表达及意义.方法 收集40例PAC、30例HGPIN和BPH标本,按常规方法石蜡包埋,制作蜡块连续切片,进行HE染色及Ki-67/caspase-3鸡尾酒抗体双重免疫组化染色.结果 Ki-67表达阳性率及增殖指数在PAC组织中明显高于HGPIN和BPH组织,且随着Gleason分级增高其增殖指数存在递增趋势(P0.05).Ki-67和caspase-3在PAC和HGPIN组织中的表达呈负相关,而在BPH中的表达呈正相关.结论 Ki-67/caspase-3在不同病变中表达不同,鸡尾酒抗体联合检测可用于前列腺良恶性疾病的诊断、鉴别诊断及预后的判断.

  14. Coarse grained modeling of transport properties in monoclonal antibody solution (United States)

    Swan, James; Wang, Gang

    Monoclonal antibodies and their derivatives represent the fastest growing segment of the bio pharmaceutical industry. For many applications such as novel cancer therapies, high concentration, sub-cutaneous injections of these protein solutions are desired. However, depending on the peptide sequence within the antibody, such high concentration formulations can be too viscous to inject via human derived force alone. Understanding how heterogenous charge distribution and hydrophobicity within the antibodies leads to high viscosities is crucial to their future application. In this talk, we explore a coarse grained computational model of therapeutically relevant monoclonal antibodies that accounts for electrostatic, dispersion and hydrodynamic interactions between suspended antibodies to predict assembly and transport properties in concentrated antibody solutions. We explain the high viscosities observed in many experimental studies of the same biologics.

  15. Comparison between Manual and Automated Methods for Ki-67 Immunoexpression Quantification in Ameloblastomas

    Directory of Open Access Journals (Sweden)

    Rogelio González-González


    Full Text Available Ameloblastoma is a common and unpredictable odontogenic tumor with high relapse rates. Several studies assessing the proliferative capacity of these neoplasms have been published, mainly using the protein Ki-67. Cell counts must be completed to determine the cell proliferation rate. Multiple methods have been developed for this purpose. The most widely used method is the labeling index, which has undergone changes over time to better facilitate cell counting. Here, we compared manual cell counting methods with automated cell counting (ImmunoRatio to determine the relative effectiveness of these methods. The results suggest that ImmunoRatio, a free software tool, may be highly advantageous and provide results similar to manual cell counting methods when used with the appropriate calibration. However, ImmunoRatio has flaws that may affect the labeling index results. Therefore, this automated cell counting method must be supplemented with manual cell counting methods.

  16. DCLK1+/Ki67- cell morphology and distribution in colorectal cancer%结直肠癌组织中DCLK1+/Ki67-肿瘤干细胞样细胞的形态和分布

    Institute of Scientific and Technical Information of China (English)

    王欢; 马法库; 刘斌; 史敏; 肖卫玲


    背景:DCLK1是在有丝分裂后的神经元中发现的一种跨膜微管相关蛋白激酶,近些年来发现其可能是肠道肿瘤干细胞的标志物。目的:观察DCLK1与Ki67在结直肠肿瘤中的表达及其病理学意义,探讨是否可以将DCLK1+/Ki67-作为结直肠癌肿瘤干细胞的标志物。方法:运用免疫组织化学方法检测150例结直肠癌组织中DCLK1与Ki67的表达,并与正常肠黏膜组织、癌旁组织、腺瘤组织比较。结果与结论:DCLK1和Ki67在癌组织中的表达率分别为36.7%,34.7%,其数量明显高于正常肠黏膜和腺瘤组织;DCLK1的表达与部位、浸润深度、淋巴结转移有关(P <0.05),而Ki67的表达仅与浸润深度有关(P <0.05);DCLK1与 Ki67的表达呈负相关(r=-0.460,P=0.000)。结直肠癌组织中DCLK1+/Ki67-细胞数量约为2.01%,主要分布于肠黏膜隐窝的基底部及共壁腺管的共壁处,细胞核大深染且呈圆形或卵圆形,核仁明显,核分裂象少见,细胞质较少。DCLK1+/Ki67-细胞在不同分化程度的腺癌组织中均存在,且分化程度越低,细胞数量越多,故无论从细胞数量和位置,还是细胞形态,都表明DCLK1+/Ki67-细胞符合肿瘤干细胞的特征,可以将DCLK1+/Ki67-作为结直肠癌肿瘤干细胞的标志物。%BACKGROUND:DCLK1 is a transmembrane microtubule-associated kinase in neurons after mitotic division, which may be the intestinal cancer stem cel marker. OBJECTIVE:To observe the expression and pathological significance of DCLK1 and Ki67 in colorectal cancer. METHODS: Expression of Ki67 and DCLK1 in 150 cases of colorectal cancer tissues was detected by immunohistochemical method in contrast to normal colorectal mucosa, para-carcinoma tissue, and adenoma tissue. RESULTS AND CONCLUSION:The expression rates of DCLK1 and Ki67 were 36.7% and 34.7% in cancer tissues, respectively, both of which were significantly higher than those in normal colorectal mucosa

  17. 乳腺多发及单发性纤维腺瘤中PTTG、Ki67的表达及意义%Expression and significance of PTTG and Ki67 in fibroadenomatosis and fibroadenomas

    Institute of Scientific and Technical Information of China (English)

    张熔熔; 黄红宇


    Objective To explore the relationship between pituitary tumor transforming gene(PTIG) and Ki67 expression and their significance in multiple adolescent breast fibroadenomas (fibroadenomatosis)and single persons(fibroadenomas) and their clinical value.Methods A total of 53 fibroadenomatosis and 160 fibroadenomas specimens in our hospital were collected.All the samples were fixed with 10% formalin and then embedded with paraffin.The protein expression of PTTG and Ki67 was detected by immunohistochemical staining.The relation between expression of PTTG and Ki67 in fibroadenomatosis was analyzed.Results The positive rate of PTTG and Ki67 was higher in fibroadenomatosis than in fibroadenomas (P < 0.005).Conclusions High expression of PTTG and Ki67 in fibroadenomatosis leads to the proliferation of tumor cell and upregulates tumor angiogenesis.High expression of PTTG and Ki67 may play important roles in tumorigenesis and pathological process of fibroadenomatosis.%目的 探讨垂体瘤转化基因(pituitary tumor transforming gene,PTTG)、细胞核增殖抗原(Ki67)表达与多发及单发性纤维腺瘤的关系及其临床意义.方法 无锡市第二人民医院2006年至2010年经手术切除的53例多发性纤维腺瘤及160例单发性纤维腺瘤标本,均经10%的福尔马林固定、石蜡包埋.运用免疫组化方法检测213例标本中PTTG、Ki67的表达情况;分析PTTG、Ki67表达与多发性、单发性纤维腺瘤的内在关系.结果 免疫组化实验结果显示PTTG、Ki67在多发性纤维腺瘤组中高表达,在单发性乳腺纤维腺瘤中有一定表达,2者差异有统计学意义(P<0.05).结论 多发性纤维腺瘤与PTTG、Ki67的高表达密切相关,2者的高表达常同时存在;PTTG、Ki67相互协同作用可促进细胞增殖,在纤维腺瘤发生或病理过程中可能起着重要的作用.

  18. Expression of S100P and Ki-67 in human benign and malignant prostatic tissue and their clinical significance%良、恶性前列腺组织中S100P与Ki-67的表达

    Institute of Scientific and Technical Information of China (English)

    李胜水; 许华; 张凤梅; 李双标; 肖丽; 刘岩; 张霖


    目的 观察人良、恶性前列腺组织中S100P与Ki-67的表达及二者的相关性.方法 应用免疫组化SP法检测15例正常前列腺组织、30例前列腺增生(prostatic hyperplasia,BPH)及30例前列腺癌(prostate cancer,PCa)组织中S100P与Ki-67的表达,并分析二者在PCa组织中的表达有无相关性.结果 (1)S100P蛋白在正常前列腺组织及BPH组织中的阳性率(93.33%、90.00%)明显高于前列腺癌组(20.00%)(χ2=38.953,P<0.01);S100P蛋白在高、中、低分化PCa组织中阳性率差异无统计学意义(P>0.05).(2)Ki-67在PCa组织中阳性率(63.33%)明显高于BPH(13.33%)及正常对照组(6.67%)(χ2=22.763,P<0.01);低分化PCa组Ki-67阳性率比高、中分化组明显增高(P<0.05).(3)PCa组织中S100P和Ki-67二者表达呈负相关关系,但无统计学意义(r=-0.077,P=0.689).结论 前列腺发生癌变后S100P与Ki-67表达分别会发生下调与升高,可能预示肿瘤更具增殖能力和侵袭性,联合检测二者有助于判断PCa的发展趋势及预后评估.%Purpose To investigate the expression of S100P protein and Ki-67 in human benign and malignant prostatic tissue, to analyze their expression correlation and clinical significance. Methods The S100P and Ki-67 expression in the tissue of fifteen normal prostate, thirty benign prostatic hyperplasia ( BPH ) patients and thirty prostate cancer ( PC a ) patients was detected by immnnohisto-chemistry. Then we analysed their relationship. Results The positive rate of S100P protein in normal and BPH groups ( 93. 33% , 90.00% ) was strikingly higher than that in the PCa tissue ( 20. 00% ) (x2 =38.953, P<0.01 ). But, there was no notable difference between the expression rate of SI OOP protein in well, moderately and poorly differentiated PCa groups ( P >0. 05 ). The expression of Ki-67 in PCa group ( 63. 33% ) was significantly higher than that in BPH ( 13.33% ) and normal group ( 6. 67% )(x2 =22.763, P< 0. 01 ). The positive rate of Ki-67 in

  19. Changes in immunoexpression of p53, Ki-67, Ets-1, APAF-1 and PTEN in serrated and conventional colon adenomas. (United States)

    Pap, Zsuzsánna; Ilyés, Izabella Ágota; Mocan, Simona Liliana; Dénes, Lóránd; Muică Nagy-Bota, Monica Cristina; Pávai, Zoltán; Szántó, Annamária


    The balance between apoptosis and proliferation is tipped towards a decrease of apoptosis as the colonocyte progresses in the adenoma to carcinoma sequence of colon carcinogenesis. According to literature data, proteins like p53, Ki-67, APAF-1, Ets-1, PTEN contribute to inhibition of apoptosis and stimulation of proliferation. Considering the complex interference among colorectal carcinogenetic mechanisms, our aim was to study the markers Ets-1 and APAF-1 relative to p53, Ki-67 and PTEN expression in colon adenomas/polyps (A/P). We performed immunohistochemistry on 99 colon A/P cases from the material of the Department of Pathology, Emergency County Hospital of Tirgu Mures, Romania. Secondary EnVision Flex/HRP (Horseradish peroxidase) (20 minutes) was used for signal amplification. The majority of A/P show increased Ki-67, p53, Ets-1 expression, decreased APAF-1 expression and preserved PTEN expression. p53, Ki-67, Ets-1 and APAF-1 demonstrated statistically significant correlations with histological type and grade of dysplasia. We also observed that expression of these proteins in the intestinal crypts has a typical distribution according to histological type and grade of dysplasia. In case of hyperplastic polyps APAF-1 expression decreases as p53 and Ki-67 expression increases, followed by a decrease in PTEN expression in serrated adenomas, and an increase of Ets-1 expression in conventional adenomas.

  20. 乳腺癌组织中 CD147和 Ki67的表达及意义

    Institute of Scientific and Technical Information of China (English)



    目的:探讨CD147、ki67与乳癌发生、发展的关系。方法采用免疫组织化学法检测78份乳癌组织中CD147蛋白、ki67表达,并分析其相关性。结果乳腺组织中CD147蛋白与ki67呈正相关( r =0469, P <0.05)。ki67阳性表达与年龄呈负相关( r =-0.469, P <0.05)。结论 CD147、ki67在乳癌中高表达,二者可能共同参与了乳癌的浸润和转移。

  1. Prognostic function of Ki-67 for pathological complete response rate of neoadjuvant chemotherapy in triple-negative breast cancer. (United States)

    Zhang, Guojing; Xie, Wanqing; Liu, Zhaozhe; Lin, Chao; Piao, Ying; Xu, Long; Guo, Fang; Xie, Xiaodong


    Triple-negative breast cancer (TNBC) has fluctuating pathological complete response (pCR) rates to neoadjuvant chemotherapy (NAC) according to published reports. Biomarkers predicting pCR rates of NAC would improve TNBC patients' outcomes. We conducted a meta-analysis to estimate the prognostic function of Ki-67 in relation to pCR rates of NAC in TNBC. Relevant publications in the literature from January 2006 to March 2013 were selected by searching PubMed, SpringerLink, Web of Science, Scopus and the Cochrane Library. The quality of prognostic studies was evaluated according to the standard reported by Hayden et al. Relative risk (RR) and 95% confidence interval (CI) were used to estimate the prognostic function of Ki-67 for pCR rates in TNBC. The fail-safe number was used to detect possible publication bias. Review Manager and MIX software was used to merge extracted data. The pCR rate of TNBC with high Ki-67 expression was 3.36 times that of low Ki-67 expression TNBC. The merged RR was 3.36 (95% CI: 1.61-7.02) and the fail-safe number was 34. No obvious publication bias but heterogeneity of the case series was detected. Ki-67 was a predictor of pCR rates to NAC in TNBC.

  2. Correlation between Ki-67 and telomerase expression with in situ hybridization for high-risk human papillomavirus

    Directory of Open Access Journals (Sweden)

    Vega-Peсa Arianna


    Full Text Available The objective of this research was to evaluate the relationship of Ki-67 and telomerase expression with the progression of cervical intraepithelial neoplasia (CIN and the physical state of the DNA of high-risk human papillomavirus (HR-HPV types. A comparative study was done on 80 biopsies of human (female cervical tissue, distributed in the following manner: 20 CIN-negative biopsies and 60 CIN-positive biopsies of varying grades. The detection of the proteins Ki-67 and telomerase was performed through immunohistochemistry; the detection of HR-HPV, by in situ hybridization. The expression of Ki-67 and telomerase increased with the progression of the CIN lesion (p <0.001. The HR-HPV genome was detected in 75% of the cases with CIN, as well as in 20% of the tissues without histological lesions (p=0.001. A significant association was found between the increase in telomerase and Ki-67 expression and the integration of the DNA of HR-HPV. The overexpression of Ki-67, telomerase and the presence the integration of the DNA of HR-HPV are evidenced by more aggressive lesions that may progress to invasive carcinoma.

  3. PROX-1 and Ki-67 expressions in cholangiocarcinoma tissues and their significance%PROX-1与Ki-67在胆管癌中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    吕品; 蒋波; 王俊; 刘剑鸣; 谭永辉



  4. Effects of cryotherapy on VEGF,Ki67 protein expression in prostate cancer of mice%冷冻对小鼠前列腺癌VEGF、Ki67表达的实验研究

    Institute of Scientific and Technical Information of China (English)

    韩艳萍; 郭志; 司同国; 王海涛; 邹强


    目的:探讨冷冻对小鼠前列腺癌血管生成及肿瘤细胞增殖的影响.方法:建立小鼠前列腺癌肿瘤模型.随机分为对照组和冷冻组,冷冻组实施氩氦冷冻治疗术.两组分别在不同时间点用免疫组化方法检测肿瘤组织的VEGF、Ki67蛋白表达.结果:对照组各时间点VEGF、Ki67蛋白表达无明显变化(P>0.05).冷冻组在冷冻后VEGF、Ki67蛋白表达均出现下降,低峰值出现在第3天,然后缓慢回升,与对照组相比,有显著性差异(P<0.01);冷冻组间VEGF、Ki67经统计分析有显著性差异(P<0.05).冷冻后前列腺癌VEGF蛋白表达水平与Ki67呈显著正相关(r=0.6921, P<0.01).结论:冷冻可明显减低肿瘤VEGF、Ki67的表达水平,可起到抑制肿瘤生长作用.

  5. The Case for Adjunctive Monoclonal Antibody Immunotherapy in Schizophrenia. (United States)

    Miller, Brian J; Buckley, Peter F


    This article presents the case in favor of clinical trials of adjunctive monoclonal antibody immunotherapy in schizophrenia. Evidence for prenatal and premorbid immune risk factors for the development of schizophrenia in the offspring is highlighted. Then key evidence for immune dysfunction in patients with schizophrenia is considered. Next, previous trials of adjunctive anti-inflammatory or other immunotherapy in schizophrenia are discussed. Then evidence for psychosis as a side effect of immunotherapy for other disorders is discussed. Also presented is preliminary evidence for adjunctive monoclonal antibody immunotherapy in psychiatric disorders. Finally, important considerations in the design and implementation of clinical trials of adjunctive monoclonal antibody immunotherapy in schizophrenia are discussed. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Preparation and Identification of Anti-rabies Virus Monoclonal Antibodies

    Institute of Scientific and Technical Information of China (English)

    Wen-juan Wang; Xiong Li; Li-hua Wang; Hu Shan; Lei Cao; Peng-cheng Yu; Qing Tang; Guo-dong Liang


    To provide a foundation for the development of rapid and specific methods for the diagnosis of rabies virus infection,anti-rabies virus monoclonal antibodies were prepared and rabies virus nucleoprotein and human rabies virus vaccine strain (PV strain) were used as immunogens to immunize 6-8 week old female BALB/c mice.Spleen cells and SP2/0 myeloma cells were fused according to conventional methods:the monoclonal cell strains obtained were selected using the indirect immunofluorescence test; this was followed by preparation of monoclonal antibody ascitic fluid; and finally,systematic identification of subclass,specificity and sensitivity was carried out.Two high potency and specific monoclonal antibodies against rabies virus were obtained and named 3B12 and 4A12,with ascitic fluid titers of 1∶8000 and 1∶10000,respectively.Both belonged to the IgG2a subclass.These strains secrete potent,stable and specific anti-rabies virus monoclonal antibodies,which makes them well suited for the development of rabies diagnosis reagents.

  7. Therapeutic monoclonal antibodies in human breast milk: a case study. (United States)

    Ross, Elle; Robinson, Steven E; Amato, Carol; McMillan, Colette; Westcott, Jay; Wolf, Tiffany; Robinson, William A


    Recently, therapeutic monoclonal antibodies have been introduced for the treatment of advanced melanoma and other diseases. It remains unclear whether these drugs can be safely administered to women who are breast feeding because of the potential hazardous side effects for nursing infants. One such therapy for metastatic melanoma is ipilimumab, a human monoclonal antibody that blocks cytotoxic T-lymphocyte-antigen-4, and is the preferred treatment for patients with metastatic melanoma when other molecular therapies are not viable. This study measured ipilimumab levels in the breast milk of a patient undergoing treatment that were enough to raise concerns for a nursing infant exposed to ipilimumab.

  8. ERBB oncogene proteins as targets for monoclonal antibodies. (United States)

    Polanovski, O L; Lebedenko, E N; Deyev, S M


    General properties of the family of tyrosine kinase ERBB receptors are considered in connection with their role in the generation of cascades of signal transduction in normal and tumor cells. Causes of acquisition of oncogene features by genes encoding these receptors and their role in tumorigenesis are analyzed. Anti-ERBB monoclonal antibodies approved for therapy are described in detail, and mechanisms of their antitumor activity and development of resistance to them are reviewed. The existing and the most promising strategies for creating and using monoclonal antibodies and their derivatives for therapy of cancer are discussed.

  9. Quantitative analysis of monoclonal antibodies by cation-exchange chromatofocusing. (United States)

    Rozhkova, Anna


    A robust cation-exchange chromatofocusing method was developed for the routine analysis of a recombinant humanized monoclonal IgG antibody. We compare the chromatofocusing method to the conventional cation-exchange chromatography (CEX) employing a salt gradient and show clear advantages of chromatofocusing over CEX. We demonstrate the suitability of the present chromatofocusing method for its intended purpose by testing the validation characteristics. To our knowledge, this is the first chromatofocusing method developed for the routine analysis of monoclonal antibody charge species.

  10. Expression and significance of CK - mit and Ki - 67 in endometrial carcinoma%线粒体肌酸激酶与Ki-67在子宫内膜癌中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    王彦英; 王淑; 周瑞民


    Objective To investigate expression and diagnostic significance of meitochondrial creatine kinase (CK - mit) and anti - Ki - 67 nuclear angigen (Ki - 67) in endometrial carcinoma.Methods Expression of CK - mit and Ki - 67 were detected in 60 cases with endometrial carcinoma and 10 cases with endometrial dysplasia and 20 cases with endometria hyperplasia by using Sp immunohisto-chemical staining. Results Ck - mit and Ki - 67 had high level expression in edometrial carcinoma. The more poorly differentiated carcinoma showed the more highly expression. The expression of Ck - mit and Ki- 67 was different significantly between endometrial carcinoma and endometrial hyperplasia (P < 0. 01) ,between endometrial dysplasia and endometrial hyperplasia(P <0.05). There wasn't significan-cy difference between endometrial dysplasia and endometrical hyperplasia. Conclusions The expression of Ck - mit and Ki - 67 indexincrease with the deceased histological differentiation in endometrial carci-noma. Ck - mit and Ki - 67 may play an important role in the development of endometrial carcinoma. The examination of Ck - mit and Ki - 67 is significant on the diagnosis, early detection and prognosis determi-nation of endometrial carcinoma.%目的 探讨子宫内膜癌细胞中线粒体肌酸激酶(CK-mit)和Kj-67核抗原抗体的表达及在子宫内膜样癌诊断中的意义.方法 采用免疫组织化学Sp法检测60例子宫内膜癌和10例非典型增生子宫内膜及20例单纯增生子宫内膜的石蜡切片组织细胞中CK-mit和Ki-67的表达.结果 CK-mit和Ki-67在子宫内膜癌中均呈高表达.两种抗体表达结果一致.癌组织分化越低,其表达越高.子宫内膜癌与非典型单纯性增殖差异无统计学意义,癌与单纯型增殖差异有统计学意义(P<0.01).非典型增殖与单纯性增殖差异有统计学意义(P<0.05).结论 CK-mit与K-67在子宫内膜癌中均呈高表达,癌组织分化程度越低,其表达越高.CK-mit和Ki-67在子宫

  11. Role of P57 and Ki-67 protein in the differential diagnosis between partial and complete hydatidiform moles%P57和Ki-67在葡萄胎鉴别诊断中的作用

    Institute of Scientific and Technical Information of China (English)

    袁雪芹; 荆福祥; 袁雪英


    Objective To investigate the role of P57 and Ki-67 protein in the differential diagnosis of partial or complete hydatidiform mole. Methods Tissues of normal placental villi, partial hydatidiform mole, and complete hydatidiform mole ( 12 for each) were collected. Distribution and expression of P57 and Ki-67 proteins were detected by immunohistochemistrical assay. Results P57 expression was located in the nuclei of villous cytotrophoblasts and stromal cells. There was no significant difference in distribution of P57 positive cells between normal villi and partial hydatidiform mole. However,none of the complete hydatidiform mole exhibited P57-positive cells. The Ki-67 expression was observed in the nuclear of cytotrophoblastic cells within placental tissues. There were significant differences in Ki-67-positive rate among normal placenta villi, partial hydatidiform mole, and complete hydatidiform mole (P<0.05). The Ki-67-positive rate in complete hydatidiform mole was higher than that in partial hydatidiform mole (P < 0.01 ). Conclusion The combined use of P57 and Ki-67 protein examination might be helpful to distinguish the complete hydatidiform mole from the partial hydatidiform mole.%目的:探讨P57和Ki-67蛋白在完全性和部分性葡萄胎鉴别诊断中的作用.方法:分别收集正常胎盘绒毛、部分性葡萄胎和完全性葡萄胎石蜡标本各12例,应用免疫组织化学方法检测P57和Ki-67蛋白在这些病变中的分布及表达水平.结果:P57蛋白在正常绒毛及部分性葡萄胎组织中主要分布于绒毛的细胞滋养叶细胞及间质细胞,两组间阳性率未见明显差异;而在完全性葡萄胎中细胞滋养叶细胞及间质细胞P57表达缺失.Ki-67蛋白主要表达于绒毛的细胞滋养叶细胞.正常绒毛组、部分性葡萄胎组和完全性葡萄胎组Ki-67阳性细胞百分率比较差别具有统计学意义(P<0.05).结论:P57结合Ki-67免疫组织化学检测可有效鉴别完全性和部分性葡萄胎.

  12. Clinicopathological and prognostic significance of Ki-67 immunohistochemical expression in gastric cancer: a systematic review and meta-analysis

    Directory of Open Access Journals (Sweden)

    Liu G


    Full Text Available Gang Liu,1,* Disheng Xiong,1,2,* Junjie Zeng,1 Borong Chen,1 Zhengjie Huang1,2 1Department of Gastrointestinal Surgery, Xiamen Cancer Hospital of the First Affiliated Hospital of Xiamen University, Xiamen, 2Department of Gastrointestinal Surgery, The First Clinical Medical College of Fujian Medical University, Fuzhou, People’s Republic of China *These authors contributed equally to this work Abstract: The prognostic significance of Ki-67 in patients with gastric cancer (GC remains controversial. The aim of our meta-analysis is to evaluate its association with clinicopathological characteristics and prognostic value in patients with GC. PubMed, EMBASE, and Web of Science were systematically searched up to May 2017. Twenty-two studies including 3,825 patients with GC were analyzed. The meta-analysis indicated that the incidence difference of Ki-67 expression in GC patients was significant when comparing the older group to younger group (odds ratio [OR] =1.44, 95% confidence interval [CI] 1.19, 1.75, lymph node positive group to negative group (OR =1.49, 95% CI 1.20, 1.84, the large size tumor group to the small size tumor group (OR =1.27, 95% CI 1.24, 1.68 and the TNM stage III+IV group to TNM stage I+II group (OR =2.28, 95% CI 1.66, 3.12. However, no statistical differences existed in gender. The detection of Ki-67 significantly correlated with the overall survival of patients (hazard ratio =1.51, 95% CI 1.31, 1.72. Our study suggested that Ki-67 overexpression was associated with poor prognosis in GC patients. Ki-67 positive rates may be associated with age, lymph node metastasis, tumor size, and TNM staging system in GC patients. Keywords: Ki-67, gastric cancer, meta-analysis, prognostic

  13. Performance of p16/Ki-67 immunostaining to detect cervical cancer precursors in a colposcopy referral population. (United States)

    Wentzensen, Nicolas; Schwartz, Lauren; Zuna, Rosemary E; Smith, Katie; Mathews, Cara; Gold, Michael A; Allen, R Andy; Zhang, Roy; Dunn, S Terence; Walker, Joan L; Schiffman, Mark


    Cytology-based screening has limited sensitivity to detect prevalent cervical precancers. Human papilloma virus (HPV) DNA testing is highly sensitive and provides a high, long-term reassurance of low risk of cervical cancer. However, the specificity of HPV DNA testing is limited, requiring additional, more disease-specific markers for efficient screening approaches. Liquid-based cytology samples were collected from 625 women referred to colposcopy. A slide was stained using the CINtec plus cytology assay. Pap cytology and HPV genotyping were conducted from the same vial. Clinical performance characteristics were calculated for all women, stratified by age, and for women referred with a low-grade squamous intraepithelial lesion (LSIL) Pap. p16/Ki-67 positivity increased with histologic severity, from 26.8% in normal histology, 46.5% in CIN1, 82.8% in CIN2 to 92.8% in CIN3. Among women with CIN3, p16/Ki-67 positivity increased from 77.8% for women younger than 30 years without HPV16 to 100% for women 30 years and older with HPV16. The sensitivity and specificity to detect CIN3+ were 93.2% and 46.1%, respectively, and increased to 97.2% and 60.0% among women 30 years and older. In women with high-risk (HR)-HPV-positive atypical squamous cells of undetermined significance (ASC-US) and LSIL, sensitivity and specificity for detection of CIN3 were 90.6% and 48.6%, respectively. p16/Ki-67 testing could reduce referral to colposcopy by almost half while detecting the most severe cases of CIN3. The high sensitivity of p16/Ki-67 with significantly improved specificity compared with HPV testing makes p16/Ki-67 a viable option for LSIL triage. Further studies are required to evaluate p16/Ki-67 as triage marker in HPV-based screening strategies.

  14. A systematic review of p16/Ki-67 immuno-testing for triage of low grade cervical cytology. (United States)

    Kisser, A; Zechmeister-Koss, I


    Screening for cervical cancer precursors by Papanicolaou cytology is a public health success story; however, its low sensitivity entails unnecessary referrals to colposcopy of healthy women with equivocal (ASCUS) or mild dysplasia (LSIL) cytology. We assessed the accuracy of p16/Ki-67 immuno-testing for triage of low grade cervical cytology. We systematically searched Medline, Embase, CRD and Cochrane databases, and handsearched key references. Eligible studies included women with ASCUS or LSIL cervical cytology who had undergone p16/Ki-67 testing and subsequent verification by colposcopy-directed biopsies and histologic analysis. We extracted data on patient characteristics and test conduct, diagnostic accuracy measures and assessed the methodological quality of the studies. R software was used to perform a bivariate analysis of test performance data. Five eligible studies were identified. Four of the studies had high risk of bias. In the LSIL subgroup, the sensitivity of p16/Ki-67 testing ranged from 0.86 to 0.98, compared with 0.92-0.96 of high-risk HPV testing (hrHPV); specificity ranged from 0.43 to 0.68 versus 0.19 to 0.37, respectively. In the ASCUS subgroup, sensitivity ranged from 0.64 to 0.92 (p16/Ki67 test) versus 0.91 to 0.97 (hrHPV); specificity ranged from 0.53 to 0.81 versus 0.26 to 0.44, respectively. p16/Ki-67 testing cannot be recommended for triage women with ASCUS or LSIL cytology due to insufficient high-quality evidence. Further studies on test performance and the impact of p16/Ki-67-based triage on health outcomes are needed for a definitive evaluation of its clinical utility. © 2014 Royal College of Obstetricians and Gynaecologists.

  15. Claudin 4 expression in triple-negative breast cancer: correlation with androgen receptors and Ki-67 expression. (United States)

    Abd-Elazeem, Mona A; Abd-Elazeem, Marwa A


    Breast cancer is the most common malignancy in women and the leading cause of cancer mortality worldwide. Triple-negative breast cancer (TNBC) is an important phenotype of breast cancer that accounts for a relatively small number of breast cancer cases but still represent a focus of increasing interest at the clinical, biological, and epidemiological level. Claudins are the major component of the tight junction, and only a few studies have addressed the role of claudins in breast cancer, especially TNBC. Androgen receptors (ARs), as members of the nuclear receptor superfamily, are known to be involved in a complex network of signaling pathways that collectively regulate cell proliferation. However, roles of AR in breast cancer development and progression have not been very clearly understood. The proliferation marker Ki-67 has been confirmed as an independent predictive and prognostic factor in early breast cancer. The aims of this study are to identify the clinicopathologic associations and prognostic value of claudin 4 expression in TNBC and to correlate claudin 4 expression with AR status and Ki-67 expression. Paraffin blocks obtained from 56 female patients with triple-negative primary invasive ductal breast carcinomas were analyzed for claudin 4, AR, and Ki-67 immunohistochemical expression. High levels of claudin 4 expression were detected in 66.1% of TNBC cases. There was a significant positive correlation with age, tumor size, grade, nodal status, metastasis, and Ki-67 expression (all P negative correlation with AR status (P negative correlation with the all the studied clinicopathologic parameters, claudin 4 and Ki-67 expression. High claudin 4 expression, negative AR expression, and high Ki-67 index would provide a strong prognostic power to differentiate the patients with worse outcome among TNBC patients. Moreover, target treatment for TNBC cells expressing claudin 4 or AR enriched would be valuable for future therapies. Copyright © 2014 Elsevier Inc

  16. Response of Triple Negative Breast Cancer to Neoadjuvant Chemotherapy: Correlation between Ki-67 Expression and Pathological Response. (United States)

    Elnemr, Gamal M; El-Rashidy, Ahmed H; Osman, Ahmed H; Issa, Lotfi F; Abbas, Osama A; Al-Zahrani, Abdullah S; El-Seman, Sheriff M; Mohammed, Amrallah A; Hassan, Abdelghani A


    Triple-negative breast cancers constitute about 15% of all cases, but despite their higher response to neoadjuvant chemotherapy, the tumors are very aggressive and associated with a poor prognosis as well as a higher risk of early recurrence. This study was retrospectively performed on 101 patients with stage II and III invasive breast cancer who received 6-8 cycles of neo-adjuvant chemotherapy. Out of the total, 23 were in the triple negative breast cancer subgroup. Nuclear Ki-67 expression in both the large cohort group (n=101) and triple negative breast cancer subgroup (n=23) and its relation to the pathological response were evaluated. The purpose of the study was to identify the predictive value of nuclear protein Ki-67 expression among patients with invasive breast cancers, involving the triple negative breast cancer subgroup, treated with neoadjuvant chemotherapy in correlation to the rate of pathological complete response. The proliferation marker Ki-67 expression was highest in the triple negative breast cancer subgroup. No appreciable difference in the rate of Ki-67 expression in triple negative breast cancer subgroup using either a cutoff of 14% or 35%. Triple negative breast cancer subgroup showed lower rates of pathological complete response. Achievement of pathological complete response was significantly correlated with smaller tumor size and higher Ki-67 expression. The majority of triple negative breast cancer cases achieved pathological partial response. The study concluded that Ki-67 is a useful tool to predict chemosensitivity in the setting of neoadjuvant chemotherapy for invasive breast cancer but not for the triple negative breast cancer subgroup.

  17. Correlation of survivin, p53 and Ki-67 in laryngeal cancer Hep-2 cell proliferation and invasion

    Institute of Scientific and Technical Information of China (English)

    Shi-Geng Pei; Ju-Xiang Wang; Xue-Ling Wang; Qing-Jun Zhang; Hong Zhang


    Objective:To investigate the mechanism of survivin, p53 and Ki-67 on Hep-2 human laryngeal cancer endothelial cell proliferation and invasion.Methods:Laryngeal squamous cell carcinoma and paracancerous normal tissues were collected, total RNA was extracted from tissues,survivin,p53and Ki-67gene mRNA expression levels in laryngeal cancer and the adjacent tissues were detected by Real-time PCR. Human laryngeal cancer Hep-2 epithelial cells were selected,survivin gene was overexpressed, and cell proliferation was detected by MTT.p53 andKi-67gene expression changes in overexpressedsurvivin gene were detected by Western blot. Changes in Hep-2 cell invasive ability were studied whensurvivin was overexpressed as detected by Transwell invasion assay.Results: In the adjacent tissues, survivin,p53andKi-67 gene relative expression levels were 1.72 ± 0.9, 13.7 ± 5.7 and 5.7 ± 1.3, respectively; while in cancer tissues, gene relative expression levels were 53.7 ± 8.3, 66.7 ± 5.2 and 61.0 ± 3.1, respectively, which was significantly increased. As detected by MTT, relative cell survival rate within 12 h ofsurvivinoverexpression were: load control group, (88.5±1.6)%; overexpressed group, (90.3±1.9)%. Transwell invasion assay results indicated that overexpressedsurvivincould significantly increase the relative survival rate of cells. Conclusions:Expressions ofp53,Ki67 and survivin are increased in cancer; and there is a positive correlation betweensurvivin, p53andKi67 expressions in laryngeal carcinoma.

  18. Correlation of survivin, p53 and Ki-67 in laryngeal cancer Hep-2 cell proliferation and invasion. (United States)

    Pei, Shi-Geng; Wang, Ju-Xiang; Wang, Xue-Ling; Zhang, Qing-Jun; Zhang, Hong


    To investigate the mechanism of survivin, p53 and Ki-67 on Hep-2 human laryngeal cancer endothelial cell proliferation and invasion. Laryngeal squamous cell carcinoma and paracancerous normal tissues were collected, total RNA was extracted from tissues, survivin, p53 and Ki-67 gene mRNA expression levels in laryngeal cancer and the adjacent tissues were detected by Real-time PCR. Human laryngeal cancer Hep-2 epithelial cells were selected, survivin gene was overexpressed, and cell proliferation was detected by MTT. p53 and Ki-67 gene expression changes in overexpressed survivin gene were detected by Western blot. Changes in Hep-2 cell invasive ability were studied when survivin was overexpressed as detected by Transwell invasion assay. In the adjacent tissues, survivin, p53 and Ki-67 gene relative expression levels were 1.72 ± 0.9, 13.7 ± 5.7 and 5.7 ± 1.3, respectively; while in cancer tissues, gene relative expression levels were 53.7 ± 8.3, 66.7 ± 5.2 and 61.0 ± 3.1, respectively, which was significantly increased. As detected by MTT, relative cell survival rate within 12 h of survivin overexpression were: load control group (88.5 ± 1.6)%; overexpressed group (90.3 ± 1.9)%. Transwell invasion assay results indicated that overexpressed survivin could significantly increase the relative survival rate of cells. Expressions of p53, Ki67 and survivin are increased in cancer; and there is a positive correlation between survivin, p53 and Ki67 expressions in laryngeal carcinoma. Copyright © 2015 Hainan Medical College. Production and hosting by Elsevier B.V. All rights reserved.

  19. Surface activity of a monoclonal antibody. (United States)

    Mahler, Hanns-Christian; Senner, Frank; Maeder, Karsten; Mueller, Robert


    The development of high concentration antibody formulations presents a major challenge for the formulation scientist, as physical characteristics and stability behavior change compared to low concentration protein formulations. The aim of this study was to investigate the potential correlation between surface activity and shaking stress stability of a model antibody-polysorbate 20 formulation. The surface activities of pure antibody and polysorbate 20 were compared, followed by a study on the influence of a model antibody on the apparent critical micelle concentration (CMC) of polysorbate 20 over a protein concentration range from 10 to 150 mg/mL. In a shaking stress experiment, the stability of 10, 75, and 150 mg/mL antibody formulations was investigated containing different concentrations of polysorbate 20, both below and above the CMC. The antibody increased significantly the apparent CMC of antibody-polysorbate 20 mixtures in comparison to the protein-free buffer. However, the concentration of polysorbate required for stabilization of the model antibody in a shaking stress experiment did not show dependence on the CMC. A polysorbate 20 level of 0.005% was found sufficient to stabilize both at low and high antibody concentration against antibody aggregation and precipitation.

  20. Expression of Bcl-2 and Ki-67 in the endometria diseases%Bcl-2和Ki-67对子宫内膜病变价值的探讨

    Institute of Scientific and Technical Information of China (English)

    文华清; 耿源源; 胡红文; 王海涛; 尹伟; 罗应彪


    Objective:To explore the expression of the Bcl-2 and Ki-67 in the endometrial. Methods:Detecting the expression of Bcl-2 and Ki-67 in the endometrial, including 25 cases of normal endometrial tissues, 150 cases of hyperplasia endometrial tissues, 75 cases of endometrial polyps tissues and 50 cases of endometrial cancer by the technology of immunohistochemistry. Results:In normal endometria, hyperplasia endometrial, endometrial polyps, endometrial cancer the posive expression rates of Bcl-2 was 84%, 62.67%, 70.67%, 38%; while those of Ki-67 was 12%, 37.33%, 17.33%, 82%. Both the expression of Bcl-2 and Ki-67 were correlated with histologic differentiation(P0.05). Conclusion:The positive expression in endometrial cancers were correlated with histologic differentiation, and it concluded that Bcl-2 and Ki-67 could be related with the endometrial cancers and normal endometrial. The expression of Bcl-2 and Ki-67 in endometrial carcinoma,in the existence of negative correlation,suggesting the application of immunohistochemical SP method for simultaneous detection of endometrial tissue in the Bcl-2,Ki-67 in abnormal protein expression, may contribute to the early detection of endometrial cancer for the early prediction of endometrial cancer and found a reference of the indicators.%目的:通过检测子宫内膜病变组织中Bcl-2和Ki-67蛋白的表达,探讨应用Bcl-2和Ki-67蛋白测定在鉴别子宫内膜病变方面的价值.方法:应用免疫组化技术SP法检测25例正常子宫内膜组织、150例子宫内膜增生症组织、75例子宫内膜息肉组织、50例子宫内膜癌组织中Bcl-2和Ki-67蛋白的表达情况.结果:Bcl-2蛋白在正常子宫内膜组织、子宫内膜增生症(单纯型、复杂型、不典型)组织、子宫内膜息肉组织、子宫内膜癌组织的阳性表达率呈递减趋势,而Ki-67的阳性表达率呈递增趋势.子宫内膜癌组织与正常子宫内膜组织、子宫内膜增生症组织、

  1. The Utility of p16INK4a and Ki-67 as a Conjunctive Tool in Uterine Cervical Lesions


    Lee, Sangho; Kim, Hyunchul; Kim, Hyesun; Kim, Chulhwan; Kim, Insun


    Background Immunohistochemical staining for p16INK4a and Ki-67 has been used to improve the accuracy in making a diagnosis of the uterine cervix cancer on biopsy. This study was conducted to examine the usefulness of these markers in the pathological diagnosis based on cervical biopsy. Methods We selected a consecutive series of 111 colposcopically directed cervical punch biopsies. Using these biopsy samples, we performed an immunohistochemical staining for p16INK4a and Ki-67 to establish a d...

  2. Antibody discovery: sourcing of monoclonal antibody variable domains. (United States)

    Strohl, William R


    Historically, antibody variable domains for therapeutic antibodies have been sourced primarily from the mouse IgG repertoire, and typically either chimerized or humanized. More recently, human antibodies from transgenic mice producing human IgG, phage display libraries, and directly from human B lymphocytes have been used more broadly as sources of antibody variable domains for therapeutic antibodies. Of the total 36 antibodies approved by major maket regulatory agencies, the variable domain sequences of 26 originate from the mouse. Of these, four are marketed as murine antibodies (of which one is a mouse-rat hybrid IgG antibody), six are mouse-human chimeric antibodies, and 16 are humanized. Ten marketed antibodies have originated from human antibody genes, three isolated from phage libraries of human antibody genes and seven from transgenic mice producing human antibodies. Five antibodies currently in clinical trials have been sourced from camelids, as well as two from non-human primates, one from rat, and one from rabbit. Additional sources of antibody variable domains that may soon find their way into the clinic are potential antibodies from sharks and chickens. Finally, the various methods for retrieval of antibodies from humans, mouse and other sources, including various display technologies and amplification directly from B cells, are described.

  3. A monoclonal antibody for G protein-coupled receptor crystallography. (United States)

    Day, Peter W; Rasmussen, Søren G F; Parnot, Charles; Fung, Juan José; Masood, Asna; Kobilka, Tong Sun; Yao, Xiao-Jie; Choi, Hee-Jung; Weis, William I; Rohrer, Daniel K; Kobilka, Brian K


    G protein-coupled receptors (GPCRs) constitute the largest family of signaling proteins in mammals, mediating responses to hormones, neurotransmitters, and senses of sight, smell and taste. Mechanistic insight into GPCR signal transduction is limited by a paucity of high-resolution structural information. We describe the generation of a monoclonal antibody that recognizes the third intracellular loop (IL3) of the native human beta(2) adrenergic (beta(2)AR) receptor; this antibody was critical for acquiring diffraction-quality crystals.

  4. Recent Progress toward Engineering HIV-1-Specific Neutralizing Monoclonal Antibodies


    Ming Sun; Yue Li; Huiwen Zheng; Yiming Shao


    The recent discoveries of broadly potent neutralizing human monoclonal antibodies represent a new generation of antiretrovirals for the treatment and prophylaxis. Antibodies are generally considered more effective and safer and have been proved to provide passive protection against mucosal challenge in humanized mice and macaques. Several neutralizing Abs could protect animals against HIV-1 but are not effective when used in an established infected model for therapy. In order to overcome the ...

  5. Expression and clinical significance of Pokemon and Ki-67 proteins in gastric cancer%Pokemon、Ki-67蛋白在胃癌中的表达和临床意义

    Institute of Scientific and Technical Information of China (English)

    韩晓峰; 李月锋; 曲宏岩


    目的 探讨Pokemon、Ki-67蛋白在胃正常黏膜与胃癌组织中的表达及其意义.方法 应用RT-PCR法检测25例胃正常黏膜及125例胃癌组织中Pokemon、Ki-67蛋白的表达,并联系临床病理学指标进行相关分析.结果 在正常胃黏膜中Pokemon和Ki-67不表达或弱表达,在胃癌组织中表达增高,并与TNM分期及有无淋巴转移有关,与年龄、性别、肿瘤大小无关.结论 Pokemon、Ki-67蛋白的表达与胃癌的发生、发展和转移密切相关,可能共同参与了胃癌的发生、发展.Pokemon有望成为胃癌治疗的一个有效靶基因,从而为胃癌基因治疗开辟新的途径.%Objective To study the expression of Pokemon、Ki -67 and its clinical significance in gastric cancer and normal gastric mucosa. Methods The Pokemon and Ki - 67 in gastric cancer(n=125), normal gastric mucosa( n = 25 )was detected hy RT - PCR. Results Pokemon protein expression was high in gastric cancer group,which was significantly different from the expression in normal gastric mucosa. It was highly expressed in gastric cancer and was related with stage of TNM and lymph node metastasis,but were not related with the age , sex and the size of tumor. Conclusion Pokemon and Ki - 67 protein expression may be correlate with the carcinogenesis and genesis, development and metastasis of gastric cancer. Pokemon may be an effective target gene for therapy of gastric cancer, accordingly , which may develop a new way of gene therapy for gastric cancer.

  6. Effect of Moxibustion on DPC4 and ki67 Expression of Pancreatic Carcinoma in Nude Mice%艾灸对胰腺癌裸鼠移植瘤DPC4、ki67表达的影响

    Institute of Scientific and Technical Information of China (English)

    仲丽丽; 孙忠人


    目的:观察艾灸对胰腺癌荷瘤裸鼠移植瘤DPC4、ki67表达影响的实验研究,并探讨艾灸抗胰腺癌的可能作用机制。方法:将16只裸鼠均接种BxPC-3细胞构建胰腺癌裸鼠移植瘤的动物模型,随机分为模型组和艾灸组。接种后次日给予麦粒灸裸鼠大椎穴至14天后结束治疗,取出肿瘤组织。采用免疫组化法检测肿瘤组织中DPC4、ki67蛋白的表达。结果:与模型组相比,艾灸组中DPC4蛋白表达升高,而ki67表达显著降低( P<0.05)。结论:艾灸对肝癌裸鼠移植瘤的生长有明显的抑制作用,其机制可能与上调肿瘤组织中DPC4、下调ki67表达水平有关。%Objective:To observe the efficacy of DPC4 and ki67 in the expression of pancreatic carcinoma in nude mice treated by moxibustion and explore the possible mechanism of antitumor through moxibustion.Meth-ods:16 nude mice were inoculated with BxPC-3 cells to construct pancreatic carcinoma xenograft models and were divided into a model group and a moxibustion group randomly.On the next day give mice moxibustion at Dazhui point.After the treatment for 14 days,the tumor tissues in nude mice were removed.Use immunohisto-chemical method to detect the protein expression of DPC4 and ki67 in tumor tissues.Results:Compared with the model group,the DPC4 expression was significantly higher and ki67 expression was lower (P<0.05).Con-clusion:Moxibustion has a significant inhibition on the growth of transplanted tumor and the mechanism may be related to the up-regulation of the expression level of DPC4 and down-regulation of the expression level of ki67 in tumor tissues.

  7. Ki-67 as a prognostic marker in mantle cell lymphoma-consensus guidelines of the pathology panel of the European MCL Network

    DEFF Research Database (Denmark)

    Klapper, W.; Hoster, E.; Determann, O.;


    powerful prognostic biomarker. The pathology panel of the European MCL Network evaluated methods to assess the Ki-67 index including stringent counting, digital image analysis, and estimation by eyeballing. Counting of 2 x 500 lymphoma cells is the gold standard to assess the Ki-67 index since this value.......37 for two methods of digital image analysis, respectively). Counting a reduced number of lymphoma cells (2 x 100 cells) showed high interobserver agreement (CCC = 0.74). Pitfalls of the Ki-67 index are discussed and guidelines and recommendations for assessing the Ki-67 index in MCL are given Udgivelsesdato......Mantle cell lymphoma (MCL) has a heterogeneous clinical course and is mainly an aggressive B cell non-Hodgkin lymphoma; however, there are some indolent cases The Ki-67 index, defined by the percentage of Ki-67-positive lymphoma cells on histopathological slides, has been shown to be a very...

  8. [Neutralizing Monoclonal and Chimeric Antibodies to Human IFN-γ]. (United States)

    Larina, M V; Aliev, T K; Solopova, O N; Pozdnyakova, L P; Korobova, S V; Yakimov, S A; Sveshnikov, P G; Dolgikh, D A; Kirpichnikov, M P


    Autoiminune disorders are chronic diseases characterized by abnormal immune response directed against self-antigens that leads to tissue damage and violation of its normal functioning. Such diseases often result in disability or even death of patients. Nowadays a number of monoclonal antibodies to pro-inflammatory cytokines and their receptors are successfully used for the targeted treatment of autoimmune diseases. One of the perspective targets in autoimmune disease therapy is interferon gamma, a key cytokine in Th1 cells differentiation, activation of macrophages, and inflammation. In the present work, 5 monoclonal antibodies to human IFN-γ were obtained. For the development of potential therapeutic agent, we have performed neutralizing activity and affinity analysis of the antibodies. Based on the data obtained, the monoclonal antibody F1 was selected. This antibody has a dissociation constant 1.7 x 10(-9) M and IC90 = 8.9 ± 2.0 nM measured upon antibody inhibition of the IFN-γ-induced HLA-DR expression on the surface of U937 cells. We have constructed a bicistronic vector for the production of recombinant chimeric Fab fragment F1 chim in E. coli cells. The recombinant chimeric Fab fragment Fl chim neutralizes IFN-γ activity in vitro and has a dissociation constant 1.8 x 10(-9) M.

  9. Heterogeneity of monoclonal antibodies revealed by charge-sensitive methods. (United States)

    Vlasak, J; Ionescu, R


    The expanding field of monoclonal antibody-based pharmaceuticals has triggered increased interest in analytical characterization of these large proteins and in understanding of their heterogeneity and degradation pathways. As a result, a large number of enzymatic modifications as well as chemical and physical degradations have been reported in monoclonal antibodies in recent years. Most heterogeneity is related to changes in the surface charge of the antibody, either directly, as a change in the number of charged residues, or indirectly as a chemical or physical alteration that changes surface-charge distribution. This review presents an overview of the sources of charge-related heterogeneity in monoclonal antibodies and the methods used for their detection. A detailed section is dedicated to deamidation of asparagine and isomerization of aspartic acid residues, two ubiquitous degradation pathways detected in antibodies and other proteins as well. Finally, kinetic modeling of the accumulation of antibody variants is presented as a tool to determine the expected fraction of molecules that have undergone one or more degradation reactions.

  10. Characterization of Binding Epitopes of CA125 Monoclonal Antibodies

    DEFF Research Database (Denmark)

    Marcos-Silva, Lara; Narimatsu, Yoshiki; Halim, Adnan


    The most used cancer serum biomarker is the CA125 immunoassay for ovarian cancer that detects the mucin glycoprotein MUC16. Several monoclonal antibodies (mAbs) including OC125 and M11 are used in CA125 assays. However, despite considerable efforts, our knowledge of the molecular characteristics...

  11. Production and potential use of monoclonal antibodies against polio viruses.

    NARCIS (Netherlands)

    A.D.M.E. Osterhaus (Albert); A.L. van Wezel; G. van Steenis (Bert); A.G. Hazendonk


    textabstractLymphocyte hybridomas secreting monoclonal antibodies against different strains of polio virus type 1, 2, or 3 have been produced. For this purpose Balb/C mice were immunized with purified and inactivated virus suspensions and their splenocytes were fused with P3X63Ag8 mouse myeloma cell

  12. Immunohistochemical diagnosis of systemic bovine zygomycosis by murine monoclonal antibodies

    DEFF Research Database (Denmark)

    Jensen, H.E.; Aalbaek, B.; Lind, Peter


    Murine monoclonal antibodies (Mabs) against water-soluble somatic antigens (WSSA) and the wall fraction (WF) from Rhizopus arrhizus (Rhizopus oryzae) were produced in vitro by fusion of splenocytes from immunized BALB/c mice with mouse myeloma X63-Ag 8.653 cells. Supernatants reacting only with h...... for the in situ diagnosis of systemic bovine zygomycosis....

  13. A mouse monoclonal antibody against Alexa Fluor 647. (United States)

    Wuethrich, Irene; Guillen, Eduardo; Ploegh, Hidde L


    Fluorophores are essential tools in molecular and cell biology. However, their application is mostly confined to the singular exploitation of their fluorescent properties. To enhance the versatility and expand the use of the fluorophore Alexa Fluor 647 (AF647), we generated a mouse monoclonal antibody against it. We demonstrate its use of AF647 for immunoblot, immunoprecipitation, and cytofluorimetry.

  14. Monoclonal antibodies for the detection of Puccinia striiformis urediniospores

    DEFF Research Database (Denmark)

    Skottrup, Peter Durand; Frøkiær, Hanne; Hearty, Stephen


    The fungal pathogen Pst causes yellow rust disease in wheat plants leading to crop losses. The organism spreads by releasing wind-dispersed urediniospores from infected plants. In this study a library of novel monoclonal antibodies (mAbs) was developed against Pst urediniospores. Nine mAb-produci...

  15. Monoclonal Antibodies to Prevent Use of Mycotoxins as Biological Weapons (United States)


    Mycotoxins as Biological Weapons PRINCIPAL INVESTIGATOR: Marta Feldmesser, M.D. CONTRACTING ORGANIZATION: Albert Einstein College of...Monoclonal Antibodies to Prevent Use of Mycotoxins as Biological Weapons 5b. GRANT NUMBER W81XWH-06-1-0085 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR

  16. Monoclonal antibodies specific for the organophosphate pesticide azinphos-methyl

    NARCIS (Netherlands)

    Jones, WT; Harvey, D; Jones, SD; Ryan, GB; Wynberg, H; TenHoeve, W; Reynolds, PHS


    2-(2-Mercapto-5-methyl-1,3,2-dioxaphosphorinan-5-yl,2-sulphide) methoxyacetic acid has been synthesized and used to prepare an azinphos hapten and protein conjugates. Monoclonal antibodies of high affinity against the pesticide azinphos-methyl were prepared from mice immunized with the

  17. Generation and Characterization of Novel Human IRAS Monoclonal Antibodies

    Directory of Open Access Journals (Sweden)

    Bo Wang


    Full Text Available Imidazoline receptors were first proposed by Bousquet et al., when they studied antihypertensive effect of clonidine. A strong candidate for I1R, known as imidazoline receptor antisera-selected protein (IRAS, has been cloned from human hippocampus. We reported that IRAS mediated agmatine-induced inhibition of opioid dependence in morphine-dependent cells. To elucidate the functional and structure properties of I1R, we developed the newly monoclonal antibody against the N-terminal hIRAS region including the PX domain (10–120aa through immunization of BALB/c mice with the NusA-IRAS fusion protein containing an IRAS N-terminal (10–120aa. Stable hybridoma cell lines were established and monoclonal antibodies specifically recognized full-length IRAS proteins in their native state by immunoblotting and immunoprecipitation. Monoclonal antibodies stained in a predominantly punctate cytoplasmic pattern when applied to IRAS-transfected HEK293 cells by indirect immunofluorescence assays and demonstrated excellent reactivity in flow immunocytometry. These monoclonal antibodies will provide powerful reagents for the further investigation of hIRAS protein functions.

  18. Monoclonal antibodies specific for the organophosphate pesticide azinphos-methyl

    NARCIS (Netherlands)

    Jones, WT; Harvey, D; Jones, SD; Ryan, GB; Wynberg, H; TenHoeve, W; Reynolds, PHS


    2-(2-Mercapto-5-methyl-1,3,2-dioxaphosphorinan-5-yl,2-sulphide) methoxyacetic acid has been synthesized and used to prepare an azinphos hapten and protein conjugates. Monoclonal antibodies of high affinity against the pesticide azinphos-methyl were prepared from mice immunized with the hapten-ovalbu

  19. Multisite phosphorylation of Pin1-associated mitotic phosphoproteins revealed by monoclonal antibodies MPM-2 and CC-3

    Directory of Open Access Journals (Sweden)

    Vincent Michel


    Full Text Available Abstract Background The peptidyl-prolyl isomerase Pin1 recently revealed itself as a new player in the regulation of protein function by phosphorylation. Pin1 isomerizes the peptide bond of specific phosphorylated serine or threonine residues preceding proline in several proteins involved in various cellular events including mitosis, transcription, differentiation and DNA damage response. Many Pin1 substrates are antigens of the phosphodependent monoclonal antibody MPM-2, which reacts with a subset of proteins phosphorylated at the G2/M transition. Results As MPM-2 is not a general marker of mitotic phosphoproteins, and as most mitotic substrates are phosphorylated more than once, we used a different phosphodependent antibody, mAb CC-3, to identify additional mitotic phosphoproteins and eventual Pin1 substrates by combining affinity purification, MALDI-TOF mass spectrometry and immunoblotting. Most CC-3-reactive phosphoproteins appeared to be known or novel MPM-2 antigens and included the RNA-binding protein p54nrb/nmt55, the spliceosomal protein SAP155, the Ki-67 antigen, MAP-1B, DNA topoisomerases II α and β, the elongation factor hSpt5 and the largest subunit of RNA polymerase II. The CC-3 mitotic antigens were also shown to be Pin1 targets. The fine CC-3- and MPM-2-epitope mapping of the RNA polymerase II carboxy-terminal domain confirmed that the epitopes were different and could be generated in vitro by distinct kinases. Finally, the post-mitotic dephosphorylation of both CC-3 and MPM-2 antigens was prevented when cellular Pin1 activity was blocked by the selective inhibitor juglone. Conclusion These observations indicate that the mitotic phosphoproteins associated with Pin1 are phosphorylated on multiple sites, suggesting combinatorial regulation of substrate recognition and isomerization.

  20. Structure and specificity of lamprey monoclonal antibodies


    Herrin, Brantley R.; Alder, Matthew N; Roux, Kenneth H.; Sina, Christina; Ehrhardt, Götz R. A.; Boydston, Jeremy A.; Turnbough, Charles L.; Cooper, Max D.


    Adaptive immunity in jawless vertebrates (lamprey and hagfish) is mediated by lymphocytes that undergo combinatorial assembly of leucine-rich repeat (LRR) gene segments to create a diverse repertoire of variable lymphocyte receptor (VLR) genes. Immunization with particulate antigens induces VLR-B-bearing lymphocytes to secrete antigen-specific VLR-B antibodies. Here, we describe the production of recombinant VLR-B antibodies specific for BclA, a major coat protein of Bacillus anthracis spores...

  1. P16INK4A及Ki-67在官颈腺癌组织中的表达及意义%Expression and significance of P16INK4A and Ki-67 in cervical adenocarcinoma

    Institute of Scientific and Technical Information of China (English)

    刘爽; 李亚里; 刘爱军; 姜淑芳; 张艾芃


    目的 探讨P 16INK4A及Ki-67蛋白在宫颈腺癌组织中的表达及其临床意义.方法 选取61例宫颈腺癌(官颈腺癌组)及34例正常官颈组织(对照组)的石蜡标本,采用SP免疫组化法测定P16INK4A及Ki-67蛋白的表达,并对61例浸润性宫颈腺癌的临床及病理资料进行回顾性分析.结果 对照组正常宫颈腺上皮P16INK4A及Ki-67表达阳性率分别为23.53%、20.59%,而宫颈腺癌组分别为80.33%、81.97%,两组比较差异有统计学意义(P<0.05).P16INK4A的表达与宫颈腺癌的临床分期、宫颈间质浸润深度、肿瘤直径有关(P<0.05),与宫颈腺癌的病理分型、肿瘤恶性程度及淋巴结转移无关(P>0.05).Ki-67的表达与宫颈腺癌的病理分化程度、肿瘤直径及淋巴结转移有关(P<0.05).子宫颈腺癌组织中P16INK4A与Ki-67的表达相关(P<0.01).结论 联合检测P16INK4A及Ki-67蛋白表达可作为宫颈腺癌的辅助诊断方法,有较好的临床应用价值.%Objective To investigate the expression and clinical implication of P16INK4A and Ki-67 in cervical adenocarcinoma. Methods In this study, 95 cases of cervical paraffin samples were divided into 2 groups: 34 cases with normal cervical glandular epithelium were designated as the control group and 61 cases with cervical adenocarcinoma tissues were designated as cervical adenocarcinoma group. SP immunohistochemistry was used to determine the expressions of P16INK4A and Ki-67. The expressions of P161NK4A and Ki-67 protein in the tissue were detected by SP immunohistochemistry, followed by qualitative and semi-quantitative determination. Furthermore, clinical and pathological data of 61 cases of invasive cervical adenocarcinoma were retrospectively analyzed. Results The positive rate of P16INIC4A and Ki-67 of the normal glandular epithelium in the control group was 23.53% and 20.59%, respectively. The positive rate of P16 and Ki-67 in the cervical adenocarcinoma group was 80.33% and 81

  2. Screening for cervical cancer precursors with p16/Ki-67 dual-stained cytology: results of the PALMS study. (United States)

    Ikenberg, Hans; Bergeron, Christine; Schmidt, Dietmar; Griesser, Henrik; Alameda, Francisco; Angeloni, Claudio; Bogers, Johannes; Dachez, Roger; Denton, Karin; Hariri, Jalil; Keller, Thomas; von Knebel Doeberitz, Magnus; Neumann, Heinrich H; Puig-Tintore, Luis M; Sideri, Mario; Rehm, Susanne; Ridder, Ruediger


    Pap cytology is known to be more specific but less sensitive than testing for human papillomavirus (HPV) for the detection of high-grade cervical intraepithelial neoplasia (CIN2+). We assessed whether p16/Ki-67 dual-stained cytology, a biomarker combination indicative of transforming HPV infections, can provide high sensitivity for CIN2+ in screening while maintaining high specificity. Results were compared with Pap cytology and HPV testing. A total of 27,349 women 18 years or older attending routine cervical cancer screening were prospectively enrolled in five European countries. Pap cytology, p16/Ki-67 immunostaining, and HPV testing were performed on all women. Positive test results triggered colposcopy referral, except for women younger than 30 years with only positive HPV test results. Presence of CIN2+ on adjudicated histology was used as the reference standard. Two-sided bias-corrected McNemar P values were determined. The p16/Ki-67 dual-stained cytology positivity rates were comparable with the prevalence of abnormal Pap cytology results and less than 50% of the positivity rates observed for HPV testing. In women of all ages, dual-stained cytology was more sensitive than Pap cytology (86.7% vs 68.5%; P p16/Ki-67 dual-stained cytology combines superior sensitivity and noninferior specificity over Pap cytology for detecting CIN2+. It suggests a potential role of dual-stained cytology in screening, especially in younger women where HPV testing has its limitations.

  3. Relationship between expression of ER, PR, Her-2, Ki-67 and neoadjuvant chemotherapy effect in breast cancer

    Institute of Scientific and Technical Information of China (English)

    Junping Xu; Hongsheng Yu


    Objective: The purpose of the study was to investigate the relationship between the expression of estrogen re-ceptor (ER), progestogen receptor (PR), human epidermal growth factor receptor (Her-2), Ki-67 and the ef ect of neoadjuvant chemotherapy in breast cancer. Methods:The expression of ER, PR, Her-2 and Ki-67 in 45 breast cancers which received neoadjuvant chemotherapy was detected by immunohistochemistry. Results:The ef ective rates in ER negative and PR negative groups were higher than those in ER positive and PR positive groups (83.3%vs 59. 4%, 82.4%vs 60.6%). There was no significant dif erence of the ef ective rate between Her-2 overexpressed group and Her-2 non-overexpressed group (81.8%vs 64.1%), and the same thing happened between Ki-67 negative group and Ki-67 positive group (67.7%vs 63.2%). Conclusion:In the patients with breast cancer, ER, PR negative ones were more sensitive to neoadjuvant chemotherapy. These patients may get more benefits from chemotherapy. ER, PR could be feasible markers for predicting the ef ective rate of neoadjuvant chemotherapy.

  4. A comparative immunohistochemical analysis of COX-2, p53, and Ki-67 expression in keratocystic odontogenic tumors

    NARCIS (Netherlands)

    Mendes, R.A.; Carvalho, J.F.C.; van der Waal, I.


    Objective. The aim of the present study was to investigate the association between the expression of cyclooxygenase-2 (COX-2) in keratocystic odontogenic tumors (KCOT) and more commonly used markers, such as p53 and Ki-67. Study design. Expression of cyclooxygenase-2 (COX-2) in 20 biopsy specimens o

  5. Low power laser irradiation stimulates cell proliferation via proliferating cell nuclear antigen and Ki-67 expression during tissue repair (United States)

    Prabhu, Vijendra; Rao, Bola Sadashiva Satish; Mahato, Krishna Kishore


    Low power laser irradiation (LPLI) is becoming an increasingly popular and fast growing therapeutic modality in dermatology to treat various ailments without any reported side effects. In the present study an attempt was made to investigate the proliferative potential of red laser light during tissue repair in Swiss albino mice. To this end, full thickness excisional wounds of diameter 15 mm created on mice were exposed to single dose of Helium-Neon laser (632.8 nm; 7 mW; 4.02 mWcm-2; Linear polarization) at 2 Jcm-2 and 10 Jcm-2 along with un-illuminated controls. The granulation tissues from all the respective experimental groups were harvested on day 10 post-wounding following euthanization. Subsequently, tissue regeneration potential of these laser doses under study were evaluated by monitoring proliferating cell nuclear antigen and Ki-67 following the laser treatment and comparing it with the un-illuminated controls. The percentages of Ki-67 or PCNA positive cells were determined by counting positive nuclei (Ki-67/PCNA) and total nuclei in five random fields per tissue sections. Animal wounds treated with single exposure of the 2 Jcm-2 indicated significant elevation in PCNA (Ptested experimental groups as evidenced by the microscopy results in the study. In summary, the findings of the present study have clearly demonstrated the regulation of cell proliferation by LPLI via PCNA and Ki-67 expression during tissue regeneration.

  6. Ki67 expression and the effect of neo-adjuvant chemotherapy on luminal HER2-negative breast cancer. (United States)

    Horimoto, Yoshiya; Arakawa, Atsushi; Tanabe, Masahiko; Sonoue, Hiroshi; Igari, Fumie; Senuma, Koji; Tokuda, Emi; Shimizu, Hideo; Kosaka, Taijiro; Saito, Mitsue


    Patients with luminal HER2-negative tumours have a favourable prognosis. However, there is a subpopulation in which poorer outcomes are obtained with endocrine therapy alone. This subpopulation is considered to benefit from chemotherapy. However, the significance of chemotherapy for those with luminal tumours has decreased due to recent changes in treatment strategies. Thus, it is often difficult to determine whether we should recommend chemotherapy to such patients in clinical practice. We investigated Ki67 expression, as a means of predicting the responses of luminal HER2-negative breast cancer patients to neo-adjuvant chemotherapy (NAC), in order to identify a subpopulation that would benefit from these treatments. We enrolled 114 luminal HER2-negative breast cancer patients undergoing surgery after NAC. Biomarkers were examined using biopsy specimens obtained prior to treatment, to avoid any chemotherapy-related effects. Chemotherapy effects were determined employing operative specimens and we defined pathological complete response (pCR) as invasive nest disappearance, based only on the primary breast tumour. We applied receiver operating characteristic curve analysis to data from our 114 patients, to investigate Ki67 expression as a predictor of pCR. The pCR rate was significantly higher for tumours with high Ki67 expression (p negative subpopulation with Ki67 expression higher than 35% benefiting from chemotherapy, as evidenced by improved survival.

  7. Analysis of human mammary fibroadenoma by Ki-67 index in the follicular and luteal phases of menstrual cycle. (United States)

    Rego, M F; Navarrete, M A L H; Facina, G; Falzoni, R; Silva, R; Baracat, E C; Nazario, A C P


    Fibroadenoma is the most common benign mammary condition among women aged 35 or younger. Expression of Ki-67 antigen has been used to compare proliferative activity of mammary fibroadenoma epithelium in the follicular and luteal phases of the menstrual cycle. Ninety eumenorrheic women were selected for tumour excision; they were assigned to either of the two groups, according to their phase of menstrual cycle. At the end of the study, 75 patients with 87 masses were evaluated by epithelial cell Ki-67 expression, blind (no information given concerning group to which any lesion belonged). Both groups were found to be homogeneous relative to age, menarche, body mass index, previous gestation, parity, breastfeeding, number of fibroadenomas, family history of breast cancer and tabagism. Median tumour size was 2.0 cm and no relationship between proliferative activity and nodule diameter was observed. No typical pattern was observed in the expression of Ki-67 in distinct nodules of the same patient. Average values for expression of Ki-67 (per 1000 epithelial cells) in follicular and luteal phases were 27.88 and 37.88, respectively (P = 0.116). Our findings revealed that proliferative activities in the mammary fibroadenoma epithelium did not present a statistically significant difference in the follicular and luteal phases. The present study contributes to clarifying that fibroadenoma is a neoplasm and does not undergo any change in the proliferative activity during the menstrual cycle.

  8. Plasma cell growth fraction using Ki-67 antigen expression identifies a subgroup of multiple myeloma patients displaying short survival within the ISS stage I. (United States)

    Gastinne, Thomas; Leleu, Xavier; Duhamel, Alain; Moreau, Anne-Sophie; Franck, Genevieve; Andrieux, Joris; Lai, Jean-Luc; Coiteux, Valerie; Yakoub-Agha, Ibrahim; Bauters, Francis; Harousseau, Jean-Luc; Zandecki, Marc; Facon, Thierry


    The current most powerful prognostic model in Multiple Myeloma (MM) combines beta-2 microglobulin (b2m) with albumin, corresponding to the International Staging System (ISS). However, the prognosis of patients within the ISS stage I (high albumin and low b2m) may vary. Ki-67 is a nuclear protein associated with cell proliferation. We retrospectively evaluated the percentage of bone marrow plasma cells expressing Ki-67 antigen (Ki-67 index) in a series of 174 untreated MM patients at diagnosis. Median survival was 51, 41 and 20 months respectively, and median Ki-67 index was 3.0%, 6.1% and 6.5% in ISS stages I, II, and III respectively. Independently of ISS, Ki-67 index > or =4% was highly predictive of adverse prognosis. Ki-67 index correlated with markers of intrinsic malignancy and with markers of tumour burden. Within ISS stage I, median survival was of 31 months (RR of death 2.65) in patients with Ki-67 index > or =4%. Eventually, the combination of Ki-67 with b2m produced an efficient prognostic model, which appeared most effective in our series when compared with b2m and KI-67 with chromosome 13 deletion models. In this series, we demonstrated that a proliferation marker provides clear-cut additional survival prognostic information to b2m into the ISS model.

  9. High‐throughput automated scoring of Ki67 in breast cancer tissue microarrays from the Breast Cancer Association Consortium (United States)

    Howat, William J; Daley, Frances; Zabaglo, Lila; McDuffus, Leigh‐Anne; Blows, Fiona; Coulson, Penny; Raza Ali, H; Benitez, Javier; Milne, Roger; Brenner, Herman; Stegmaier, Christa; Mannermaa, Arto; Chang‐Claude, Jenny; Rudolph, Anja; Sinn, Peter; Couch, Fergus J; Tollenaar, Rob A.E.M.; Devilee, Peter; Figueroa, Jonine; Sherman, Mark E; Lissowska, Jolanta; Hewitt, Stephen; Eccles, Diana; Hooning, Maartje J; Hollestelle, Antoinette; WM Martens, John; HM van Deurzen, Carolien; Investigators, kConFab; Bolla, Manjeet K; Wang, Qin; Jones, Michael; Schoemaker, Minouk; Broeks, Annegien; van Leeuwen, Flora E; Van't Veer, Laura; Swerdlow, Anthony J; Orr, Nick; Dowsett, Mitch; Easton, Douglas; Schmidt, Marjanka K; Pharoah, Paul D; Garcia‐Closas, Montserrat


    Abstract Automated methods are needed to facilitate high‐throughput and reproducible scoring of Ki67 and other markers in breast cancer tissue microarrays (TMAs) in large‐scale studies. To address this need, we developed an automated protocol for Ki67 scoring and evaluated its performance in studies from the Breast Cancer Association Consortium. We utilized 166 TMAs containing 16,953 tumour cores representing 9,059 breast cancer cases, from 13 studies, with information on other clinical and pathological characteristics. TMAs were stained for Ki67 using standard immunohistochemical procedures, and scanned and digitized using the Ariol system. An automated algorithm was developed for the scoring of Ki67, and scores were compared to computer assisted visual (CAV) scores in a subset of 15 TMAs in a training set. We also assessed the correlation between automated Ki67 scores and other clinical and pathological characteristics. Overall, we observed good discriminatory accuracy (AUC = 85%) and good agreement (kappa = 0.64) between the automated and CAV scoring methods in the training set. The performance of the automated method varied by TMA (kappa range= 0.37–0.87) and study (kappa range = 0.39–0.69). The automated method performed better in satisfactory cores (kappa = 0.68) than suboptimal (kappa = 0.51) cores (p‐value for comparison = 0.005); and among cores with higher total nuclei counted by the machine (4,000–4,500 cells: kappa = 0.78) than those with lower counts (50–500 cells: kappa = 0.41; p‐value = 0.010). Among the 9,059 cases in this study, the correlations between automated Ki67 and clinical and pathological characteristics were found to be in the expected directions. Our findings indicate that automated scoring of Ki67 can be an efficient method to obtain good quality data across large numbers of TMAs from multicentre studies. However, robust algorithm development and rigorous pre‐ and post

  10. p16(INK4a) /Ki-67 co-expression specifically identifies transformed cells in the head and neck region. (United States)

    Prigge, Elena-Sophie; Toth, Csaba; Dyckhoff, Gerhard; Wagner, Steffen; Müller, Franziska; Wittekindt, Claus; Freier, Kolja; Plinkert, Peter; Hoffmann, Jürgen; Vinokurova, Svetlana; Klussmann, Jens Peter; von Knebel Doeberitz, Magnus; Reuschenbach, Miriam


    p16(INK4a) immunohistochemical overexpression is an overall reliable surrogate marker of human papillomavirus (HPV)-associated head and neck squamous cell carcinomas (HNSCC). However, cases of ambiguous p16(INK4a) overexpression are regularly detected in the head and neck: p16(INK4a) expression can be observed in non-malignant tissue, such as tonsillar crypt epithelium and a proportion of branchial cleft cysts. Additionally, diverse patterns of p16(INK4) expression can complicate interpretation of "p16(INK4a) -positivity". These aspects impede the unrestricted application of p16(INK4a) as a diagnostic marker in the head and neck. We hypothesized that combined detection of p16(INK4a) and the proliferation marker Ki-67 could support clarification of ambiguous p16(INK4a) expression in the head and neck by specifically indicating p16(INK4a) -expressing cells with proliferative activity. p16(INK4a) /Ki-67 co-expression in a combined staining procedure was correlated to distinct p16(INK4a) expression patterns and HPV status (HPV DNA followed by E6*I oncogene mRNA detection) in 147 HNSCC and 50 non-malignant head and neck samples. p16(INK4a) /Ki-67 co-expression only occurred in transformed cells of the head and neck. Co-expression was never detected in non-transformed cells. Combined p16(INK4a) /Ki-67 expression was stringently associated with a diffuse p16(INK4a) expression pattern. All HPV oncogene-expressing HNSCC showed p16(INK4a) /Ki-67 co-expression. We demonstrate that p16(INK4a) /Ki-67 co-expression occurs exclusively in transformed cells of the head and neck. Our findings indicate a substantial impact of combined p16(INK4a) /Ki-67 expression in the assessment of ambiguous p16(INK4a) expression in the head and neck by specifically identifying p16(INK4a) -expressing cells with proliferative activity. This property will be of considerable significance for head and neck histo- and cytopathology.

  11. Development of a Ki-67-based clinical trial assay for neoadjuvant endocrine therapy response monitoring in breast cancer. (United States)

    Goncalves, Rodrigo; DeSchryver, Katherine; Ma, Cynthia; Tao, Yu; Hoog, Jeremy; Cheang, Maggie; Crouch, Erika; Dahiya, Neha; Sanati, Souzan; Barnes, Michael; Sarian, Luis Otávio Zanatta; Olson, John; Allred, Donald Craig; Ellis, Matthew J


    The recent publication of the ACOSOG Z1031 trial results demonstrated that Ki-67 proliferation marker-based neoadjuvant endocrine therapy response monitoring could be used for tailoring the use of adjuvant chemotherapy in ER+HER2-negative breast cancer patients. In this paper, we describe the development of the Ki-67 clinical trial assay used for this study. Ki-67 assay assessment focused on reproducing a 2.7% Ki-67 cut-point (CP) required for calculating the Preoperative Endocrine Prognostic Index and a 10% CP for poor endocrine therapy response identification within the first month of neoadjuvant endocrine treatment. Image analysis was assessed to increase the efficiency of the scoring process. Clinical outcome concordance for two independent Ki-67 scores was the primary performance metric. Discordant scores led to a triage approach where cases with complex histological features that software algorithms could not resolve were flagged for visual point counting (17%). The final Ki-67 scoring approach was run on T1/2 N0 cases from the P024 and POL trials (N = 58). The percent positive agreement for the 2.7% CP was 87.5% (95% CI 61.7-98.5%); percent negative agreement 88.9% (95% CI: 65.3-98.6%). Minor discordance did not affect the ability to predict similar relapse-free outcomes (Log-Rank P = 0.044 and P = 0.055). The data for the 10% early triage CP in the POL trial were similar (N = 66), the percentage positive agreement was 100%, and percent negative agreement 93.55% (95% CI: 78.58-99.21%). The independent survival predictions were concordant (Log-rank P = 0.0001 and P = 0.01). We have developed an efficient and reproducible Ki-67 scoring system that was approved by the Clinical Trials Evaluation Program for NCI-supported neoadjuvant endocrine therapy trials. Using the methodology described here, investigators are able to identify a subgroup of patients with ER+HER2-negative breast cancer that can be safely managed without the need of adjuvant

  12. Development of Biodegradable Nanocarriers Loaded with a Monoclonal Antibody

    Directory of Open Access Journals (Sweden)

    Andrew Gdowski


    Full Text Available Treatments utilizing monoclonal antibody therapeutics against intracellular protein-protein interactions in cancer cells have been hampered by several factors, including poor intracellular uptake and rapid lysosomal degradation. Our current work examines the feasibility of encapsulating monoclonal antibodies within poly(lactic-co-glycolic acid (PLGA nanoparticles using a water/oil/water double emulsion solvent evaporation technique. This method can be used to prepare protective polymeric nanoparticles for transporting functional antibodies to the cytoplasmic compartment of cancer cells. Nanoparticles were formulated and then characterized using a number of physical and biological parameters. The average nanoparticle size ranged from 221 to 252 nm with a low polydispersity index. Encapsulation efficiency of 16%–22% and antibody loading of 0.3%–1.12% were observed. The antibody molecules were released from the nanoparticles in a sustained manner and upon release maintained functionality. Our studies achieved successful formulation of antibody loaded polymeric nanoparticles, thus indicating that a PLGA-based antibody nanoformulation is a promising intracellular delivery vehicle for a large number of new intracellular antibody targets in cancer cells.

  13. Significado clínico-patológico das expressões citofotométricas do Ki-67 e Caspase-3 no carcinoma de células escamosas do esôfago Clinicopathologic significance of the Ki-67 and Caspase-3 cytophotometric expressions in the esophageal squamous cell carcinomal

    Directory of Open Access Journals (Sweden)

    Gilmar Pereira Silva


    se mostraram in-tensas sendo que a da Caspase-3 foi superior ao Ki-67 mas sem correlação com as características clínico-patológicas.BACKGROUND: The esophageal squamous cell carcinoma treatment strategy is still based on the tumor staging, where tumor histopathologic charac-teristics are the major determinants. In parallel, studies have been developed in order to better understand the tumor biology using immunohistochemical meth-ods with manual quantification evaluating the proliferative and apoptotic activi-ties of the cells. The disadvantages related to the manual method rose the de-velopment of computerized ways to do the image analysis. OBJETIVES: To verify the expressions of the markers Ki-67 (proliferative and Caspase-3 (apoptotic and to correlate them with the clinic and pathologic characteristics of the tumor. METHODS: Twenty-nine paraffin embedded blocks were studied, each one con-taining tissue samples from patients with esophageal squamous cell carcinoma submitted to esophagectomies. The clinic and pathological data were obtained from histopathologic informations and from medical records. The slides were prepared following the routine immunohistochemical method until the point to utilize the specific antibodies (MIB-1 and CPP32. Positive quantification of the immunoreactivity to the proteins Ki-67 and Caspase-3 was performed by the software for computerized image analysis SAMBA (Systeme d' Analyse Micro-photometrique a Balayage Automatique. Statistical analysis was done having P3cm; and lesions located in the lower third of the organ. The mean score indexes found were 62.05% for Ki-67 and 86.06% for Caspase-3 and there was no correlation with the clinic or pathologi-cal characteristics as gender, age and tumor staging. There was significant dif-ference of Ki-67 expression among the histological grades (P=0.047 and corre-lation between the evaluated indexes (r=0.41 and P=0.032. CONCLUSION: The protein expressions were high and the Caspase-3 protein

  14. p16、Ki67、PHH3和HPV-L1在宫颈脱落细胞中的表达及临床意义%Expression and clinical significance of p16, Ki67, PHH3 and HPV-L1 in exfoliated cervical cells

    Institute of Scientific and Technical Information of China (English)

    黄平; 张雪; 陈光元; 金瑛


    目的:观察p16基因、Ki67蛋白、核分裂特异性抗体(PHH3)及人乳头瘤病毒L1型(HPV-L1)基因在宫颈病变脱落细胞的表达规律,并探讨其临床意义。方法选取2015年4月至2016年4月期间本院妇科收治的100例妇科病患者作为研究对象,取宫颈脱落细胞,行杂交捕获Ⅱ代法人乳头瘤病毒(HPV HC-Ⅱ)检测和液基薄层细胞学检查,检测并分析HPV-L1、PHH3、Ki67、p16的表达水平。结果宫颈脱落细胞学检出未见上皮内细胞病变(NILM)8例,非典型鳞状上皮细胞/低级别鳞状上皮内病变(ASCUS/LSIL)42例,非典型鳞状上皮细胞不能排除高级别鳞状上皮内病变/高级别鳞状上皮内病变(ASC-H/HSIL)41例,鳞状细胞癌(SCC)9例。ASCUS/LSIL患者的Ki67、PHH3、p16阳性检出率分别为19.05%、4.76%、26.19%,均明显低于ASC-H/HSIL患者的85.37%、88.89%、88.89%,差异均有统计学意义(P<0.05);p16阳性表达对宫颈癌及癌前病变的诊断敏感性、特异度分别为90.32%(56/62)、86.84%(33/38),Ki67分别为83.87%(52/62)、89.47%(34/38),PHH3分别为70.97%(44/62)、100.00%(38/38);ASCUS/LSIL患者的HPV-L1阳性表达率为69.05%,明显高于ASC-H/HSIL (0),差异有显著统计学意义(P<0.01)。结论 PHH3、Ki67、p16联合检测可作为筛查宫颈脱落细胞的辅助手段,其能够有效提高宫颈癌及癌前病变的诊断准确性,检测HPV-L1能够对ASCUS/SLIL病变程度进行有效分流,为临床治疗提供一定的指导。%Objective To observe the expression of p16 gene and Ki67 protein, mitosis-specific antibody anti-phosphohistone-H3 (PHH3) and human papilloma virus (HPV) L1 protein (HPV-L1) gene in exfoliated cervical epithelial cells for cervical lesions, and to explore its clinical significance. Methods A total of 100 patients of gy-necological disease in Department of Gynecology from April 2015 to April 2016 were selected as research subjects. The exfoliated cervical cells were

  15. The use of combinations of monoclonal antibodies in clinical oncology. (United States)

    Henricks, Linda M; Schellens, Jan H M; Huitema, Alwin D R; Beijnen, Jos H


    Treatment with monoclonal antibodies is becoming increasingly important in clinical oncology. These antibodies specifically inhibit signaling pathways in tumor growth and/or induce immunological responses against tumor cells. By combining monoclonal antibodies several pathways may be targeted simultaneously, potentially leading to additive or synergistic effects. Theoretically, antibodies are very suitable for use in combination therapy, because of limited overlapping toxicity and lack of pharmacokinetic interactions. In this article an overview is given of preclinical and clinical data on twenty-five different combinations of antibodies in oncology. Some of these combinations have proven clinical benefit, for example the combination of trastuzumab and pertuzumab in HER2-positive breast cancer, which exemplifies an additive or synergistic effect on antitumor activity in clinical studies and the combination of nivolumab and ipilimumab, which results in significant increases in progression-free and overall survival in patients with advanced melanoma. However, other combinations may lead to unfavorable results, such as bevacizumab with cetuximab or panitumumab in advanced colorectal cancer. These combinations result in shorter progression-free survival and increased toxicity compared to therapy with a single antibody. In summary, the different published studies showed widely varying results, depending on the combination of antibodies, indication and patient population. More preclinical and clinical studies are necessary to unravel the mechanisms behind synergistic or antagonistic effects of combining monoclonal antibodies. Most research on combination therapies is still in an early stage, but it is expected that for several tumor types the use of combination therapy of antibodies will become standard of care in the near future.

  16. [Single B cell monoclonal antibody technologies and applications]. (United States)

    Chi, Xiangyang; Yu, Changming; Chen, Wei


    Monoclonal antibodies (mAbs) contribute a lot to the development of numerous fields in life science as a pivotal tool in modern biological research. Development of the PCR methods and maturation of antibody production have made it possible to generate mAbs from single human B cells by single cell RT-PCR with successional cloning and expression in vitro. Compared to traditional monoclonal antibody technologies, single B cell technologies require relatively fewer cells, which are highly efficient in obtaining specific mAbs in a rapid way with preservation of the natural heavy and light chain pairing. With so many advantages, single B cell technologies have been proved to be an attractive approach for retrieval of naive and antigen-experienced antibody repertoires generated in vivo, design of rationale structure-based vaccine, evaluation and development of basic B cell biology concepts in health and autoimmunity, and prevention of infectious diseases by passive immunization and therapy for disorders. Accordingly, this review introduced recent progresses in the single B cell technologies for generating monoclonal antibodies and applications.

  17. Library of monoclonal antibodies against brush border membrane epithelial antigens

    Energy Technology Data Exchange (ETDEWEB)

    Behar, M.; Katz, A.; Silverman, M.


    A purified fraction of proximal tubule brush border membranes (BBM) was prepared from dog kidney and used to immunize mice. The standard technique of hybridoma production was followed as described by Kohler and Milstein. Production of antibodies was detected by indirect immunofluorescence on dog kidney slices and by immunodot against the purified fraction on nitrocellulose. Five hybrids exhibited anti BBM activity. These were cloned twice and yielded stable cell lines producing IgG type monoclonal antibodies against BBM. They were designated A/sub 1/, C/sub 7/, D/sub 3/, D/sub 7/ and H/sub 4/. As a family these five monoclonals have broad tissue specificity, i.e. positive staining of the surface mucosa of intestinal kidney proximal tubules. D/sub 3/ exhibits even broader specificity for epithelium reacting with bile canaliculi and choroid plexus. The authors have verified that at least 4/5 antibodies are directed against BBM protein as revealed by immunoprecipitation of solubilized BBM and detected by Coomassie blue staining or autoradiography of lactoperoxidase labelled BBM. Most interestingly all antibodies bind to the surface of LL CPK/sub 1/ cells, a continuous pig kidney cell line of undefined origin but exhibiting many characteristics of proximal tubule cells. The library of monoclonal antibodies obtained provide important probes with which to study membrane biogenesis and polarization in epithelial cells.

  18. Adsorption of monoclonal antibodies to glass microparticles. (United States)

    Hoehne, Matthew; Samuel, Fauna; Dong, Aichun; Wurth, Christine; Mahler, Hanns-Christian; Carpenter, John F; Randolph, Theodore W


    Microparticulate glass represents a potential contamination to protein formulations that may occur as a result of processing conditions or glass types. The effect of added microparticulate glass to formulations of three humanized antibodies was tested. Under the three formulation conditions tested, all three antibodies adsorbed irreversibly at near monolayer surface coverages to the glass microparticles. Analysis of the secondary structure of the adsorbed antibodies by infrared spectroscopy reveal only minor perturbations as a result of adsorption. Likewise, front-face fluorescence quenching measurements reflected minimal tertiary structural changes upon adsorption. In contrast to the minimal effects on protein structure, adsorption of protein to suspensions of glass microparticles induced significant colloidal destabilization and flocculation of the suspension.

  19. Skp2 expression is associated with high risk and elevated Ki67 expression in gastrointestinal stromal tumours

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    Terracciano Luigi


    Full Text Available Abstract Background Gastrointestinal stromal tumors (GIST exhibit an unpredictable clinical course and can rapidly progress to lethality. Predictions about the biological behavior of GIST are based on a number of canonical clinical and pathologic parameters whose validity in distinguishing between a benign and a malignant tumour is still imperfect. The aim of our study was to investigate the role of morphologic parameters and expression of cells cycle regulators as prognosticators in GIST. Methods We performed an immunohistochemical analysis for Ki67, p27Kip1, Jab1, and Skp2, on a Tissue Microarray (TMA containing 94 GIST. Expression of the above proteins was correlated to classically used prognosticators, as well as to risk groups. Clinical significance of histologic and immunohistochemical features were evaluated in 59 patients for whom follow-up information was available. Results Overexpression of Ki67 and Skp2, and p27Kip1 loss directly correlated with the high risk group (p = 0.03 for Ki67 and Skp2, p = 0.05 for p27Kip1. Jab1 expression did not exhibit correlation with risk. In 59 cases provided with clinical follow-up, high cellularity, presence of necrosis, and Ki67 overexpression were predictive of a reduced overall survival in a univariate model. The same parameters, as well as mitotic rate, tumour size, and p27Kip1 loss were indicative of a shortened relapse free survival interval. High cellularity, and high mitotic rate retained their prognostic significance by multivariate analysis. Conclusion Our data suggest that a number of histologic parameters in combination with immunohistochemical expression of cell cycle regulators can facilitate risk categorization and predict biologic behavior in GIST. Importantly this study demonstrates, for the first time, that Skp2 expression correlates with Ki67 expression and high risk in GIST.

  20. Importance of P53, Ki-67 expression in the differential diagnosis of benign/malignant phyllodes tumors of the breast

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    Ulku Kucuk


    Full Text Available Background: Conventionally growth pattern, stromal overgrowth, stromal cellularity and stromal mitotic activity are the main parameters in the grading of phyllodes tumors (PTs. Recent studies revealed that both p53 and Ki-67 expressions are correlated with grade of PTs of the breast. Expression of hormone receptors and overexpression/amplification of HER2 has been studied in PTs to discover the roles of these markers as new treatment modalities. Materials and Method: We studied 26 PT cases. Seventeen benign and nine malignant PTs were re-evaluated as regards stromal cellularity mitotic activity, p53/Ki-67 expression rates and the relation between these parameters. Estrogen receptor and progesterone receptor (ER, PR positivity were determined by counting nuclear staining in five high-power fields. Also, the presence of any HER2 staining and staining patterns were documanted. Results: Stromal cellularity, mitotic rate, p53 and Ki-67 expression rates were all correlated with benign and malignant histologic subgroups (P = 0.000-0.001. Ki-67 and p53 expressions were statistically significantly correlated with histologic subgroups, stromal cellularity and mitotic rate (P < 0.005. ER and PR expressions in the epithelial component were not statistically significant between the two groups. HER2 showed different staining patterns in the epithelial component, and there was no staining in the stromal component. Conclusion: Ki-67 and p53 expression rates were statistically significantly correlated with grade of mammary PTs; therefore, they can be used in the determination of tumor grade, especially for the differential diagnosis of benign and malignant tumors. Malignant and benign tumors did not differ significantly in terms of hormone receptor and HER2 expression. HER2 expression showed different patterns in the epithelial component of the PTs.

  1. Evaluation of Ki67, p16 and CK17 Markers in Differentiating Cervical Intraepithelial Neoplasia and Benign Lesions

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    Fatemeh Sari Aslani


    Full Text Available Background: Cervical intraepithelial neoplasia (CIN is a premalignant lesion capable of progressing to cervical cancer. Despite the existing well-defined criteria, the histomorphologic diagnosis is subject to high rates of discordance among pathologists. The aim of this study was to evaluate Ki-67 (MIB-1, CK17 and p16 INK4a (p16 markers by immunohistochemical methods in differentiating CIN from benign cervical lesions. Methods: The present study reviewed and re-classified 77 cervical biopsies, originally diagnosed as 31 non-CIN, and 46 CIN, as 54 non-CIN, and 23 CIN based on at least two similar diagnoses. Immunostaining by Ki67, p16 and CK17 markers was performed on all cases and the results were compared with pervious and consensus diagnosis.Results: The overall agreement between pervious and consensus diagnosis was 67.5% (Kappa=0.39, P<0.001. The sensitivity and specificity of Ki67 immunostaining were 95.6% and 85.1% respectively, while for p16 the corresponding values were 91.3% and 98.1%. The overall agreement, for both p16 and Ki67, with consensus diagnosis were significant (P<0.001. The sensitivity and specificity of CK17 negative staining in CIN detection were 39.1% and 40.7% respectively.Conclusion: Ki67 and p16 markers are recommended as complementary tests for differentiating between dysplastic and non-dysplastic lesions. CK17 does not discriminate between immature metaplasia with and without dysplasia.

  2. [P16(INK4alpha)/Ki-67 immunocytochemical dual staining for detection of cervical lesions associated to papillomavirus infection]. (United States)

    Toro de Méndez, Morelva; Ferrández Izquierdo, Antonio; Llombart-Bosch, Antonio


    We aimed to explore the expression pattern of p16(INK4alpha)/Ki-67 immunocytochemical dual-staining and to establish the potential clinical utility for early detection of cervical lesions. Liquid-based cytologies of cervical specimens of cervical cancer screening were processed for p16(INK4alpha)/Ki-67 immunocytochemical dual-staining using the CINtec Plus Kit. HPV testing was performed with the INNO-LiPA HPV genotyping Extra Reverse Hybridization Line Probe Assay kit. One hundred and fifteen cervical cytologies were analyzed with the following results: 11(9.6%) were negative for intraepithelial lesions or malignancy (NILM); 32 (27.8%) presented atypical squamous cells of undetermined significance (ASC-US); 62 (53.9%) exhibited low grade squamous intraepithelial lesions (LSIL) and 10 (8.7%) showed high grade squamous intraepithelial lesions (HSIL). No cases of cervical cancer were detected. The overall prevalence of DNA HPV detection was 81.7% (94/115). The following specific HPV genotypes were identified in 42 (45.0%) cases: HPV16 (26.2%), HPV51 (21.4%), HPV52 (14.3%) and HPV66 (7.1%). Viral sequences of an unknown single HPV were detected in 23.8% of the cases. A total of 42/115 (36.5%) were p16(IVK4alpha)/Ki-67 dual-staining-positive, being more frequent in HSIL (70.0%), decreasing in LSIL (44.0%), detected in a minority of ASC-US (25.0%) and negative in NILM cases (p p16(INK4alpha)/Ki-67 dual-staining, including 6/32 (18.8%) ASC-US, 26/62 (42.0%) LSIL and 8/10 (80.0%) HSIL, which represent a strong association between positivity for HPV, p16(INK4alpha)/Ki-67 staining and severe cytological abnormalities (p cervical lesions.

  3. 乳腺癌的超声BI-RADS与Ki-67表达的相关性

    Institute of Scientific and Technical Information of China (English)

    郑琼; 李胜利; 陈琮瑛; 王文韬; 梁冠男; 袁鹰; 余蓉


    目的探讨应用乳腺影像学报告及数据系统(BI-RADS)描述乳腺癌超声指标与Ki-67表达的相关性及其临床意义。方法对174名在南方医科大学附属深圳妇幼保健院乳腺外科住院治疗并经病理证实的乳腺癌患者共计194个乳腺癌病灶的超声BIRADS指标与免疫组织化学方法测定的肿瘤Ki-67表达结果进行对比分析。结果 194个乳腺癌病灶中,超声诊断乳腺癌的敏感度为86.1%,Ki-67阳性表达123例,阳性表达率63.4%(123/194)。单因素分析显示Ki-67阳性表达的乳腺癌肿块中边缘成角、边缘毛刺、内部微钙化、血流丰富指标出现比率高于阴性肿块(P<0.05)。多因素分析显示边缘毛刺和血流丰富进入回归模型(P<0.05)。结论乳腺癌超声BI-RADS指标与Ki-67表达具有一定的相关性,可根据乳腺癌的超声表现初步预测其预后,为临床治疗方案的选择提供一定的帮助。

  4. Expression of Ki-67, p53, and K-ras in chronic pancreatitis and pancreatic ductal adenocarcinoma

    Institute of Scientific and Technical Information of China (English)

    Seok Jeong; Young Bae Kim; Don Haeng Lee; Jung Il Lee; Jin-Woo Lee; Kye Sook Kwon; Pum-Soo Kim; Hyung Gil Kim; Yong Woon Shin; Young Soo Kim


    AIM: To examine surgical specimens of pancreas with either chronic pancreatitis or pancreatic cancer in order to study whether ductal hyperplasia and dysplasia in pancreas represent precursor lesions for pancreatic cancer.METHODS: We examined expression of Ki-67, CEA,p53, and K-ras, in the surgical specimens of pancreas with adenocarcinomas (n = 11) and chronic pancreatitis (n = 12). Cellular proliferation was assessed by Ki-67proliferation index using the proliferation marker Ki-67.In specimens with pancreas cancer, we divided pancreas epithelium into normal (n=7), ductal hyperplasia (n=3), dysplasia (n=4), and cancerous lesion (n=11) after hematoxylin and eosin staining, Ki-67, and CEA immunohistochemical staining. In cases with chronic pancreatitis, the specimen was pathologically examined as in cases with pancreas cancer, and they were also determined as normal (n=10), ductal hyperplasia (n=4), or dysplasia (n= 5). p53 and K-ras expression were also studied by immunohistochemical staining.RESULTS: In pancreatic cancer, the Ki-67 index was 3.73±3.58 in normal site, 6.62±4.39 in ductalhyperplasia, 13.47±4.02 in dysplasia and 37.03±10.05in cancer tissue, respectively. Overall, p53 was positive in normal ducts, ductal hyperplasia, dysplasia, and carcinoma cells in 0 of 14 (0%), 0 of 7 (0%), 7 of 9 (78%),and 10 of 11 (91%), respectively, and K-ras was positive in 0 of 8 (0%), 1 of 3 (33%), 4 of 6 (67%), 4 of 5 (80%),respectively.CONCLUSION: Our results favorably support the hypothesis that ductal hyperplasia and dysplasia of the pancreas might be precursor lesions for pancreas cancer.Further evaluation of oncogenes by the molecular study is needed.

  5. Predictive value of the combined p16 and Ki-67 immunocytochemistry in low-grade squamous intraepithelial lesions. (United States)

    Ziemke, Peter; Marquardt, Katrin; Griesser, Henrik


    The reliability of cytological diagnoses, especially for low-grade squamous intraepithelial lesions (LSIL), is limited. This leads to uncertainty in patient management. The application of adjunctive biomarkers is meant to improve this situation. Therefore, we examined the prognostic value of p16/Ki-67 immunostaining of LSIL cytology specimens. We analyzed the p16(INK4a) and Ki-67 immunocytochemistry (CINtec® PLUS, dual stain) of 260 patients with LSIL. Cytology and dual-stain results were correlated with histology at the time of treatment or with cytological follow-up. After an average duration of 24.9 months (1-58) and a histology rate of 36.2% [cervical intraepithelial neoplasia, grade 2 or higher (CIN2+) as positive], the statistical evaluation for cytology and dual stain resulted in a sensitivity of 98.3 and 90.0%, respectively, a specificity of 74.5% for dual stain, a positive predictive value (PPV) of 22.8 and 51.4%, and a negative predictive value (NPV) of 96.1% for dual stain. The combined immunocytochemical investigation of p16(INK4a) and Ki-67 leads to a significantly better PPV and a very good NPV for CIN2+ in LSIL, especially in women 30 years of age and older. An objective individualized prognosis may not be achieved with p16(INK4a)/Ki-67. Statistical data from our study, however, indicate that patient management can be significantly improved by the application of combined p16/Ki-67 immunocytochemistry as an adjunct to cytology. © 2014 S. Karger AG, Basel.

  6. Monoclonal IgA Antibodies for Aflatoxin Immunoassays



    Antibody based techniques are widely used for the detection of aflatoxins which are potent toxins with a high rate of occurrence in many crops. We developed a murine monoclonal antibody of immunoglobulin A (IgA) isotype with a strong binding affinity to aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2) and aflatoxin M1 (AFM1). The antibody was effectively used in immunoaffinity column (IAC) and ELISA kit development. The performance of the IACs was compatible ...

  7. Sperm-immobilizing monoclonal antibody to human seminal plasma antigens. (United States)

    Shigeta, M; Watanabe, T; Maruyama, S; Koyama, K; Isojima, S


    Rat spleen cells immunized to human azoospermic semen (a mixture of seminal plasma components) and mouse myeloma cells (P3/X63 Ag8U1; P3U1) (Marguilies et al., 1976) were successfully fused with polyethylene glycol (PEG 1500) and 19 of 89 fused cell cultures were found to produce sperm-immobilizing antibody. The cells that produced antibody indicating the highest sperm-immobilizing activity were distributed into wells for further recloning and 10 clones producing sperm-immobilizing antibody were established. The clone (1C4) producing the highest antibody titre was found to produce a large amount of IgG in culture supernatants and to contain a mixture of rat and mouse chromosomes. It was proved by immunodiffusion test that the monoclonal antibody was produced to the human seminal plasma antigen No. 7 which is common to human milk protein. Using this hybridoma which produced a large amount of monoclonal sperm-immobilizing antibody, a new method could be developed for purifying human seminal plasma antigen by immunoaffinity chromatography with bound antibody from the hybridoma. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:6783353

  8. Generation of monoclonal antibodies against highly conserved antigens.

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    Hongzhe Zhou

    Full Text Available BACKGROUND: Therapeutic antibody development is one of the fastest growing areas of the pharmaceutical industry. Generating high-quality monoclonal antibodies against a given therapeutic target is very crucial for the success of the drug development. However, due to immune tolerance, some proteins that are highly conserved between mice and humans are not very immunogenic in mice, making it difficult to generate antibodies using a conventional approach. METHODOLOGY/PRINCIPAL FINDINGS: In this report, the impaired immune tolerance of NZB/W mice was exploited to generate monoclonal antibodies against highly conserved or self-antigens. Using two highly conserved human antigens (MIF and HMGB1 and one mouse self-antigen (TNF-alpha as examples, we demonstrate here that multiple clones of high affinity, highly specific antibodies with desired biological activities can be generated, using the NZB/W mouse as the immunization host and a T cell-specific tag fused to a recombinant antigen to stimulate the immune system. CONCLUSIONS/SIGNIFICANCE: We developed an efficient and universal method for generating surrogate or therapeutic antibodies against "difficult antigens" to facilitate the development of therapeutic antibodies.

  9. Monoclonal IgA Antibodies for Aflatoxin Immunoassays

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    Özlem Ertekin


    Full Text Available Antibody based techniques are widely used for the detection of aflatoxins which are potent toxins with a high rate of occurrence in many crops. We developed a murine monoclonal antibody of immunoglobulin A (IgA isotype with a strong binding affinity to aflatoxin B1 (AFB1, aflatoxin B2 (AFB2, aflatoxin G1 (AFG1, aflatoxin G2 (AFG2 and aflatoxin M1 (AFM1. The antibody was effectively used in immunoaffinity column (IAC and ELISA kit development. The performance of the IACs was compatible with AOAC performance standards for affinity columns (Test Method: AOAC 991.31. The total binding capacity of the IACs containing our antibody was 111 ng, 70 ng, 114 ng and 73 ng for AFB1, AFB2, and AFG1 andAFG2, respectively. Furthermore, the recovery rates of 5 ng of each AF derivative loaded to the IACs were determined as 104.9%, 82.4%, 85.5% and 70.7% for AFB1, AFB2, AFG1 and AFG2, respectively. As for the ELISA kit developed using non-oriented, purified IgA antibody, we observed a detection range of 2–50 µg/L with 40 min total test time. The monoclonal antibody developed in this research is hitherto the first presentation of quadruple antigen binding IgA monoclonal antibodies in mycotoxin analysis and also the first study of their utilization in ELISA and IACs. IgA antibodies are valuable alternatives for immunoassay development, in terms of both sensitivity and ease of preparation, since they do not require any orientation effort.

  10. Monoclonal IgA Antibodies for Aflatoxin Immunoassays (United States)

    Ertekin, Özlem; Pirinçci, Şerife Şeyda; Öztürk, Selma


    Antibody based techniques are widely used for the detection of aflatoxins which are potent toxins with a high rate of occurrence in many crops. We developed a murine monoclonal antibody of immunoglobulin A (IgA) isotype with a strong binding affinity to aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2) and aflatoxin M1 (AFM1). The antibody was effectively used in immunoaffinity column (IAC) and ELISA kit development. The performance of the IACs was compatible with AOAC performance standards for affinity columns (Test Method: AOAC 991.31). The total binding capacity of the IACs containing our antibody was 111 ng, 70 ng, 114 ng and 73 ng for AFB1, AFB2, and AFG1 andAFG2, respectively. Furthermore, the recovery rates of 5 ng of each AF derivative loaded to the IACs were determined as 104.9%, 82.4%, 85.5% and 70.7% for AFB1, AFB2, AFG1 and AFG2, respectively. As for the ELISA kit developed using non-oriented, purified IgA antibody, we observed a detection range of 2–50 µg/L with 40 min total test time. The monoclonal antibody developed in this research is hitherto the first presentation of quadruple antigen binding IgA monoclonal antibodies in mycotoxin analysis and also the first study of their utilization in ELISA and IACs. IgA antibodies are valuable alternatives for immunoassay development, in terms of both sensitivity and ease of preparation, since they do not require any orientation effort. PMID:27187470

  11. Monoclonal IgA Antibodies for Aflatoxin Immunoassays. (United States)

    Ertekin, Özlem; Pirinçci, Şerife Şeyda; Öztürk, Selma


    Antibody based techniques are widely used for the detection of aflatoxins which are potent toxins with a high rate of occurrence in many crops. We developed a murine monoclonal antibody of immunoglobulin A (IgA) isotype with a strong binding affinity to aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2) and aflatoxin M1 (AFM1). The antibody was effectively used in immunoaffinity column (IAC) and ELISA kit development. The performance of the IACs was compatible with AOAC performance standards for affinity columns (Test Method: AOAC 991.31). The total binding capacity of the IACs containing our antibody was 111 ng, 70 ng, 114 ng and 73 ng for AFB1, AFB2, and AFG1 andAFG2, respectively. Furthermore, the recovery rates of 5 ng of each AF derivative loaded to the IACs were determined as 104.9%, 82.4%, 85.5% and 70.7% for AFB1, AFB2, AFG1 and AFG2, respectively. As for the ELISA kit developed using non-oriented, purified IgA antibody, we observed a detection range of 2-50 µg/L with 40 min total test time. The monoclonal antibody developed in this research is hitherto the first presentation of quadruple antigen binding IgA monoclonal antibodies in mycotoxin analysis and also the first study of their utilization in ELISA and IACs. IgA antibodies are valuable alternatives for immunoassay development, in terms of both sensitivity and ease of preparation, since they do not require any orientation effort.

  12. DAPI nuclear stain and Ki-67 expression in atypical thyroid adenoma and their clinical significance%甲状腺非典型腺瘤中DAPI核染色、Ki-67表达及意义

    Institute of Scientific and Technical Information of China (English)

    邢益祥; 孟刚


    目的 探讨4',6-二脒基-2-苯基吲哚(4',6-diamidino-2-phenylindole,DAPI)细胞核染色、Ki-67在甲状腺非典型腺瘤(atypical thyroid adenoma,ATA)中的表达及临床意义.方法 采用免疫组化EnVision两步法和免疫荧光染色法分别检测ATA、甲状腺滤泡癌(follicular thyroid carcinoma,FC)中Ki-67增殖指数及DAPI核染色情况.结果 DAPI在低、高核级ATA组织中的阳性率分别为12.50% (2/16)、16.13%(5/31),两组相比差异无统计学意义(x2=0.01,P>0.05);其在对照组FC中的阳性率为85.71%(6/7),两组相比差异有统计学意义(x2=13.17,P<0.01);Ki-67在低、高核级ATA组织中的阳性率分别为6.25%(1/16)、6.45%(2/31),二者相比差异无统计学意义(x2 =0.36,P>0.05);而在对照组FC中的阳性率为85.71% (6/7),二者相比差异有统计学意义(x2=22.19,P<0.01);Ki-67、DAPI表达与ATA患者年龄、结节数量、肿块大小无关.结论 联合检测Ki-67和DAPI可能对ATA与FC的鉴别诊断有一定价值.Ki-67、DAPI与ATA核级别、患者年龄、肿块大小和结节数量无关.

  13. Generation of HER2 monoclonal antibodies using epitopes of a rabbit polyclonal antibody. (United States)

    Hu, Francis Jingxin; Uhlen, Mathias; Rockberg, Johan


    One of the issues in using polyclonal antibodies is the limited amount of reagent available from an immunisation, leading to batch-to-batch variation and difficulties in obtaining the same antibody performance when the same antigen is re-immunised into several separate animals. This led to the development of hybridoma technology allowing, at least theoretically, for an unlimited production of a specific binder. Nevertheless, polyclonal antibodies are widely used in research and diagnostics and there exists a need for robust methods to convert a polyclonal antibody with good binding performance into a renewable monoclonal with identical or similar binding specificity. Here we have used precise information regarding the functional recognition sequence (epitope) of a rabbit polyclonal antibody with attractive binding characteristics as the basis for generation of a renewable mouse monoclonal antibody. First, the original protein fragment antigen was used for immunisation and generation of mouse hybridoma, without obtaining binders to the same epitope region. Instead a peptide designed using the functional epitope and structural information was synthesised and used for hybridoma production. Several of the monoclonal antibodies generated were found to have similar binding characteristics to those of the original polyclonal antibody. These monoclonal antibodies detected native HER2 on cell lines and were also able to stain HER2 in immunohistochemistry using xenografted mice, as well as human normal and cancer tissues.

  14. Proliferation Determined by Ki-67 Defines Different Pathologic Response to Neoadjuvant Trastuzumab-Based Chemotherapy in HER2-Positive Breast Cancer. (United States)

    Sánchez-Muñoz, Alfonso; Navarro-Perez, Victor; Plata-Fernández, Yessica; Santonja, Angela; Moreno, Ignacio; Ribelles, Nuria; Alba, Emilio


    This study aimed to assess the role of proliferation measured by Ki-67 as a predictive factor for pathologic complete response (pCR) to trastuzumab-based chemotherapy in patients with human epidermal growth factor receptor 2 (HER2)-positive (HER2(+)) breast cancer (BC). A total of 81 patients with HER2(+) BC were treated with a sequential schedule consisting of 4 cycles of cyclophosphamide (600 mg/m(2)) and doxorubicin (60 mg/m(2)) every 3 weeks, followed by 4 cycles of weekly paclitaxel (80 mg/m(2)) or docetaxel (100 mg/m(2)) every 3 weeks combined with trastuzumab (loading dose of 8 mg/kg and then 6 mg/kg every 3 weeks) as neoadjuvant treatment. Histologic subgroups classified by hormone receptor (HR) expression and Ki-67 index were 17% HR(+)/Ki-67 ≥ 50%, 41% HR(+)/Ki-67 < 50%, 25% HR-negative (HR(-)) Ki-67 ≥ 50%, and 17% HR(-)/Ki-67 < 50%. pCR, defined as the absence of invasive cells in the breast and axillary lymph node, was achieved in 33 patients (41%). The median Ki-67 expression was significantly higher in tumors with pCR (53%) compared with tumors without pCR (30%) (P < .001). Receiver operating characteristic (ROC) curve methodology suggested that 50% was the optimal Ki-67 cutoff point to best identify patients who achieved a pCR. The pCR rate was significantly different between histologic subgroups: HR(-)/Ki-67 ≥ 50% (70%), HR(+)/Ki-67 ≥ 50% (71%), HR(-)/Ki-67 < 50% (22%), and HR(+)/Ki-67 < 50% (18%) (P < .001). A multivariate analysis revealed that a Ki-67 marker ≥ 50% was the only independent predictive factor of pCR (P = .003; odds ratio [OR], 0.133; 95% confidence interval [CI], 0.036-0.5). The median follow-up was 32 months (range, 14-48 months). Patients who achieved a pCR had significantly lower recurrence (P = .001) and higher overall survival (OS) (P = .013) compared with those who did not. There were no statistically significant differences in disease-free survival (DFS) and OS in relation to HRs, the Ki-67 marker as a continuous or

  15. The Use of Monoclonal Antibodies in Human Prion Disease (United States)

    Bodemer, Walter

    Detection of PrP and its pathological isoform(s) is the key to understanding the etiology and pathogenesis of transmissible spongiform encephalopathy. There is ample evidence that PrP isoforms constitute a major component of an unknown and perhaps unconventional infectious agent. An etiological relationship between human and zoonotic transmissible spongiform encephalopathies may be revealed with monoclonal antibodies. Knowledge of the conformational transition rendering a nonpathogenic, almost ubiquitous cellular protein into a pathogenic one is crucial to defining pathomechanisms. The stepwise or even continuous formation of pathogenic molecules can be monitored. Any improvement in the early diagnosis could help to conceive new therapeutic measures which are not currently available. Determination of PrP isoforms in tissue, cells, or body fluids may be of prognostic value. Many experimental approaches in molecular medicine and molecular biology of the prion protein already rely on monoclonal antibodies. Recombinant antibodies such as the single-chain Fv may soon replace traditional hybridoma techniques. Binding affinity can easily be manipulated by a number of techniques, including in vitro mutagenesis - a step which could never be carried out using the traditional hybridoma technology. Monoclonal antibodies are and will remain an essential support for ongoing research on the prion protein in general and on the unconventional infectious prions.

  16. Challenges and opportunities for monoclonal antibody therapy in veterinary oncology. (United States)

    Beirão, Breno C B; Raposo, Teresa; Jain, Saurabh; Hupp, Ted; Argyle, David J


    Monoclonal antibodies (mAbs) have come to dominate the biologics market in human cancer therapy. Nevertheless, in veterinary medicine, very few clinical trials have been initiated using this form of therapy. Some of the advantages of mAb therapeutics over conventional drugs are high specificity, precise mode of action and long half-life, which favour infrequent dosing of the antibody. Further advancement in the field of biomedical sciences has led to the production of different forms of antibodies, such as single chain antibody fragment, Fab, bi-specific antibodies and drug conjugates for use in diagnostic and therapeutic purposes. This review describes the potential for mAbs in veterinary oncology in supporting both diagnosis and therapy of cancer. The technical and financial hurdles to facilitate clinical acceptance of mAbs are explored and insights into novel technologies and targets that could support more rapid clinical development are offered. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. A recombinant, fully human monoclonal antibody with antitumor activity constructed from phage-displayed antibody fragments

    NARCIS (Netherlands)

    Huls, GA; Heijnen, IAFM; Cuomo, ME; Koningsberger, JC; Boel, E; de Vries, ARV; Loyson, SAJ; Helfrich, W; Henegouwen, GPV; van Meijer, M; de Kruif, J; Logtenberg, T


    A single-chain Fv antibody fragment specific for the tumor-associated Ep-CAM molecule was isolated from a semisynthetic phage display library and converted into an intact, fully human IgG1 monoclonal antibody (huMab), The purified huMab had an affinity of 5 nM and effectively mediated tumor cell kil

  18. Evaluation of p16/Ki-67 dual staining in detection of cervical precancer and cancers: a multicenter study in China. (United States)

    Yu, Lu-Lu; Chen, Wen; Lei, Xiao-Qin; Qin, Yu; Wu, Ze-Ni; Pan, Qin-Jing; Zhang, Xun; Chang, Bai-Feng; Zhang, Shao-Kai; Guo, Hui-Qin; Qiao, You-Lin


    To analyze the clinical performance of p16/Ki-67 dual-stained cytology identifying high-grade cervical intraepithelial neoplasia (CIN2+) in Chinese women. 1079 women attending ongoing cervical cancer screening and 211 "enriched" women aged ≥30yrs with biopsy-confirmed CIN2+ from five Chinese hospitals were enrolled during year 2014-2015. Cervical specimens were collected for high-risk human papillomavirus (HR-HPV) DNA analysis, Liquid-based cytology (LBC) and p16/Ki-67 dual staining. Colposcopy and biopsy were performed on women with any abnormal result. p16/Ki-67 positivity increased with histologic severity. It was 18.4%(183/996) in normal histology, 54.0%(34/63) in CIN1, 81.0%(34/42) in CIN2, 93.3%(111/119) in CIN3, 71.4% (5/7) in adenocarcinoma and 95.2%(60/63) in squamous cell carcinoma. Compared with the HR-HPV negatives, p16/Ki-67 expression was significantly higher in the HPV16/18 positive (OR: 35.45(95%CI: 23.35-53.84)) and other 12 HR-HPV types positive group (OR: 8.01(95%CI: 5.81-11.05). The sensitivity and specificity of p16/Ki-67 to detect CIN2+ in the entire population were 90.9% and 79.5%, respectively. In women with ASC-US and LSIL, sensitivity and specificity for detection of CIN2+ were 87.5% and 66.4%, respectively, with a referral rate of 43.8%. In women who tested positive for HR-HPV, sensitivity and specificity of dual-staining for detection of CIN2+ were 92.7% and 52.7%, respectively, and the referral rate was 68.7%. p16/Ki-67 dual-stained cytology provided a high sensitivity and moderate specificity to detect underlying cervical precancer and cancers in various settings, and might be considered as an efficient screening tool in China.

  19. Current status of cancer immunodetection with radiolabeled human monoclonal antibodies. (United States)

    De Jager, R; Abdel-Nabi, H; Serafini, A; Pecking, A; Klein, J L; Hanna, M G


    The use of radiolabeled murine monoclonal antibodies (MoAbs) for cancer immunodetection has been limited by the development of human antimouse antibodies (HAMA). Human monoclonal antibodies do not elicit a significant human antihuman (HAHA) response. The generation and production of human monoclonal antibodies met with technical difficulties that resulted in delaying their clinical testing. Human monoclonal antibodies of all isotypes have been obtained. Most were immunoglobulin (Ig) M directed against intracellular antigens. Two antibodies, 16.88 (IgM) and 88BV59 (IgG3k), recognize different epitopes on a tumor-associated antigen, CTA 16.88, homologous to cytokeratins 8, 18, and 19. CTA 16.88 is expressed by most epithelial-derived tumors including carcinomas of the colon, pancreas, breast, ovary, and lung. The in vivo targeting by these antibodies is related to their localization in nonnecrotic areas of tumors. Repeated administration of 16.88 over 5 weeks to a cumulative dose of 1,000 mg did not elicit a HAHA response. Two of 53 patients developed a low titer of HAHA 1 to 3 months after a single administration of 88BV59. Planar imaging of colorectal cancer with Iodine-131 (131I)-16.88 was positive in two studies in 9 of 12 and 16 of 20 patients preselected by immunohistochemistry. Tumors less than 2 cm in diameter are usually not detected. The lack of immunogenicity and long tumor residence time (average = 17 days) makes 16.88 a good candidate for therapy. Radioimmunlymphoscintigraphy with indium-111 (111In)-LiLo-16.88 administered by an intramammary route was used in the presurgical staging of primary breast cancer. The negative predictive value of lymph node metastases for tumors less than 3 cm was 90.5%. Planar and single photon emission computed tomography imaging of colorectal carcinoma with technetium-99m (99mTc) 88BV59 was compared with computed tomography (CT) scan in 36 surgical patients. The antibody scan was more sensitive than the CT scan in detecting

  20. Immunohistological expression of HIF-1α, GLUT-1, Bcl-2 and Ki-67 in consecutive biopsies during chemoradiotherapy in patients with rectal cancer

    DEFF Research Database (Denmark)

    Havelund, Birgitte Mayland; Sørensen, Flemming Brandt; Pløen, John;


    receiving preoperative CRT (>50.4 Gy and Uracil/Tegafur). Immunohistological expressions of HIF-1α, GLUT-1, Bcl-2 and Ki-67 were investigated in biopsies taken before treatment, after 2, 4 and 6 weeks of CRT and in specimens from the operation. Decreasing expressions of HIF-1α, Bcl-2 and Ki-67 were observed...

  1. Evaluation of p16/Ki-67 dual-stained cytology as triage test for high-risk human papillomavirus-positive women

    NARCIS (Netherlands)

    Ebisch, R.M.F.; Horst, J.; Hermsen, M.; Rijstenberg, L.L.; Vedder, J.E.; Bulten, J.; Bosgraaf, R.P.; Verhoef, V.M.; Heideman, D.A.; Snijders, P.J.; Meijer, C.J.W.; Kemenade, F.J. van; Massuger, L.F.; Melchers, W.J.; Bekkers, R.L.M.; Siebers, A.G.


    The aim of this study was to evaluate the clinical utility of p16/Ki-67 dual staining, for the identification of CIN in high-risk HPV-positive women from a non-responder screening cohort. P16/Ki-67 dual staining, Pap cytology, and HPV16/18 genotyping were performed on physician-taken liquid-based sa

  2. 宫颈癌组织中ADAM-19、Ki-67的表达及临床意义%The expression and clinical significance of ADAM-19,Ki-67 in cervical cancer

    Institute of Scientific and Technical Information of China (English)

    逯仁波; 王小川; 马荣; 耿晓星


    目的 研究早期宫颈癌组织中去整合素基质金属蛋白酶-19(a disintegrin and metalloproteinase,ADAM-19)、癌细胞增殖指数(Ki-67)的表达及临床意义.方法 应用免疫组化SP法检测18例正常宫颈上皮(normal cervical epithelium,NCE)、22例宫颈上皮内瘤变(cervical intraepithelial neoplasm,CIN)和82例宫颈早期浸润癌(invasive cervix carsinoma,ICC)组织中ADAM-19和Ki-67的表达情况.结果 在宫颈癌中ADAM-19表达于癌细胞浆和或细胞膜;Ki-67表达于细胞核.从正常宫颈上皮(normal cervical epithelium,NCE)到宫颈上皮内瘤变(cervical intraepithelial neoplasm,CIN)再到宫颈浸润癌(invasive carsinoma cervix,ICC),ADAM-19、Ki-67的阳性表达率逐步升高(P<0.05).ADAM-19在宫颈浸润癌中的表达与盆腔淋巴结转移、脉管浸润、间质浸润、国际妇产科联盟(FIGO)分期、组织学分级和Ki-67表达有关(P<0.05);但与年龄和组织学类型无明显相关性(P>0.05).有盆腔淋巴结转移、脉管浸润、突破深层间质浸润、FIGO分期为Ⅱ期、组织学分级为Ⅲ级及Ki-67高度表达者,其ADAM-19阳性表达率显著高于无盆腔淋巴结转移、无脉管浸润、浸润深度在浅层间质以内、FIGO分期为Ⅰ期、组织学分级未超过Ⅱ级及Ki-67表达在中度以内者(P<0.05).结论 ADAM-19阳性表达可能在癌细胞增殖和侵袭转移中起重要作用.ADAM-19过度表达者,癌细胞增殖活跃,更易发生侵袭转移,但并非唯一决定因素.检测宫颈癌中ADAM-19表达对进一步了解宫颈癌生物学行为和判断其预后有一定价值.%Objective To evaluate the expression and clinical significance of A disintegrin and metalloproteinase( ADAM - 19 ),Ki - 67 in early invasive cervical cancer. Methods Expression of ADAM - 19 and Ki - 67 in 18 cases of normal cervical epithelium( NCE ),22 cases of cervical intraepithelial neoplasm( CIN ) and 82 cases of invasive carcinoma cervix( ICC )were detected by

  3. Ki67与核酸荧光共染在细胞周期研究中的应用%Application of Ki67 and nuclear acid fluorescent dye co-staining in analysis of cell cycle

    Institute of Scientific and Technical Information of China (English)



    病原微生物与宿主细胞的相互作用是感染过程中的一个重要环节,也是研究肿瘤生物学的一项重要内容。动态研究细胞周期变化对了解病原体作用于细胞具有重要意义。本研究用Ki67/4′,6‐二脒基‐2‐苯基吲哚(4′,6‐diamidino‐2‐phenylindole ,DAPI)和Ki67/碘化丙啶(propidium iodide ,PI)共染技术分析了小鼠脾细胞的细胞周期变化,并介绍其具体应用。%The interaction between pathogenic microbes and host cells is crucial for studies on infections and tumor cell biology . The kinetic studies of cell cycle are of significance for understanding the mechanisms mediated by microbes versus host cells .In this paper ,a novel method for cell cycle analysis in mouse splenic cells by Ki67 and 4′,6‐diamidino‐2‐phenylindole (DAPI) or propidium iodide (PI) co‐staining is introduced .

  4. P53和Ki67在胃癌中的表达及其临床意义%Clinical significance of P53 and Ki67 expression in gastric cancer

    Institute of Scientific and Technical Information of China (English)

    刘伟; 余英豪; 欧阳学农; 王烈; 武一曼; 陈娟; 熊喜生


    AIM: To investigate the correlation of P53 and Ki67 protein expression with the clinicopathological parameters in gastric cancer.METHODS: A total of 1 107 surgical specimens of gastric cancer were collected from the Fuzhou General Hospital of Nanjing Military Command,The expression of P53 and Ki67 was examined by immunohistochemistry in 775 gastric cancer specimens taken from patients with complete clinical data.RESULTS: The positive rates of P53 and Ki67 protein expression were 46.84% and 78.97%, respectively. Higher positive rates of P53 and Ki67 protein expression were noted in intestinal-type gastric cancer than in diffuse-type gastric cancer (54.7% vs 38.0%, 86.0% vs 70.0%, both P < 0.05)and in well-differentiated cancer than in poorlydifferentiated cancer (54.7 vs 35.4%, 87.0% vs 67.2%, both P < 0.05). A higher positive rate of P53 protein expression was noted in gastroesophageal junction (GEJ) cancer than in gastric cancer (53.7% vs 41.9%, P < 0.05) and in cancer without distant metastasis than in that with distant metastasis (49.0% vs 30.3%, P < 0.05). A higher positive rate of Ki67 protein expression was noted in invasive cancer than in less invasive cancer ( 80.3% vs 60.4%, P < 0.05) and in cancer with lymph node metastasis than in that without lymph node metastasis (81.4% vs 72.6 %, P < 0.05).CONCLUSION: P53 and Ki67 protein expression might be associated with the development, invasion, and metastasis of gastric cancer, and can be used to evaluate the malignant behavior and prognosis of gastric cancer.%目的:探讨胃癌中P53和Ki67蛋白表达与临床病理参数的相关性及其临床意义.方法:收集在中国人民解放军南京军区福州总医院手术切除的胃癌组织标本1 107例,采用EliVision plus免疫组织化学染色方法检测其中临床资料完整的775例胃腺癌标本中Ki67结果:在775例胃腺癌组织标本中,P53和Ki67蛋白的阳性表达率分别为46.84%和78.97%,

  5. Production of monoclonal antibodies to human glomerular basement membrane.

    Directory of Open Access Journals (Sweden)



    Full Text Available Using the technique of somatic cell fusion, we produced monoclonal antibodies to collagenase-digested human glomerular basement membrane (GBM. Fourteen monoclonal antibodies which reacted with normal human kidney in indirect immunofluorescence (IIF studies were produced. An analysis of the binding patterns indicated that the antigens recognized could be divided into six broad groups. Monoclonal antibody B3-H10 (Group 1 reacted with only GBM in a fine granular pattern. A5-B12 and B5-C2 (Group 2 reacted with GBM and peritubular capillary in a linear pattern. B2-A12 (Group 3 reacted with only epithelial cells. Al-C9 and A4-E2 (Group 4 showed a mesangial pattern in glomerulus and a lineal pattern in tubular basement membrane (TBM, Bowman's capsule and peritubular capillary. A1-E1, A1-E11, A2-E6, A3-B6, A4-F8 and B5-H2 (Group 5 recognized determinants common to GBM, TBM, Bowman's capsule and/or peritubular capillary. A3-F1 and B5-E10 (Group 6 reacted with TBM and Bowman's capsule. The staining pattern of B3-H10 (Group 1 was characteristic because it was not linear, but finely granular along the GBM. The staining pattern of B2-A12 (Group 3 was also characteristic because only epithelial cells were stained, and processes of epithelial cells were observed as fine fibrils. To the best of our knowledge, these two types of monoclonal antibodies have not been reported previously.

  6. Strategies for Treating Autoimmune Disease With Monoclonal Antibodies


    Wofsy, David


    There is no safe and reliable therapy for most serious autoimmune diseases, such as systemic lupus erythematosus. Severe cases usually require treatment with corticosteroids or cytotoxic drugs or both, which frequently provide inadequate disease control and can cause serious complications. These therapies are not restricted in their effects to cells of the immune system, but rather have a broad range of toxic effects on cells throughout the body. The development of monoclonal antibodies has l...

  7. Quantification of Moraxella bovis haemagglutinating adhesins with monoclonal antibodies. (United States)

    Gil-Turnes, C; Aleixo, J A


    Six monoclonal antibodies (MAbs) against Moraxella bovis GF 9 were used to quantify haemagglutinating adhesins of 16 strains of this organism. The amount of each MAb necessary to inhibit one haemagglutinating unit of each strain varied between 4 and 0.007 times that required by strain GF 9. Five strains reacted with six MAbs, one with five, two with four, one with three, two with two and three with none. The procedures used enabled to detect dominant strains candidates for vaccines.

  8. Monoclonal antibody to native P39 protein from Borrelia burgdorferi.


    Sullivan, T J; Hechemy, K E; Harris, H L; Rudofsky, U H; Samsonoff, W A; Peterson, A J; Evans, B. D.; Balaban, S L


    We have produced, by using a sonicate of Borrelia burgdorferi, a monoclonal antibody (MAb), NYSP39H, that is specific for the P39 protein band. This MAb reacted with 13 isolates of B. burgdorferi but not with eight different spirochetes (four borrelias, two leptospiras, and two treponemas). Surface labeling of B. burgdorferi with biotin and subsequent treatment with Nonidet P-40 showed that P39 was not biotinylated but was extracted with Nonidet P-40, indicating that it is present within the ...

  9. Monoclonal Antibody-Based Therapeutics for Melioidosis and Glanders

    Directory of Open Access Journals (Sweden)

    Hyung-Yong Kim


    Full Text Available Problem statement: Burkholderia Pseudomallei (BP and B. Mallei (BM were two closely related pathogenic gram-negative bacteria. They were the causative agents of melioidosis and glanders, respectively and are recognized by CDC as category B select agents. Significant efforts had been devoted to developing the diagnostic and therapeutic measures against these two pathogens. Monoclonal antibody-based therapeutic was a promising targeted therapy to fight against melioidosis and glanders. Valuable findings have been reported by different groups in their attempt to identify vaccine targets against these two pathogens. Approach: Our group has generated neutralizing Monoclonal Antibodies (MAbs against BP and BM and characterized them by both in vitro and in vivo experiments. We present an overview of the MAb-based therapeutic approaches against BP and BM and demonstrate some of our efforts for developing chimeric and fully human MAbs using antibody engineering. Results: Throughout conventional mouse hybridoma technique and antibody engineering (chimerization and in vitro antibody library techniques, we generated 10 chimeric MAbs (3 stable MAbs and 7 transient MAbs and one fully human MAb against BP and BM. In addition, we present the reactive antigen profiles of these MAbs. Our approaches had potentials to accelerate the development of therapeutics for melioidosis and glanders in humans. Conclusion: Our experience and findings presented here will be valuable for choosing the best antigenic targets and ultimately for the production of effective vaccines for these two pathogens.

  10. Nucleolus disassembly in mitosis and apoptosis: dynamic redistribution of phosphorylated-c-Myc, fibrillarin and Ki-67

    Directory of Open Access Journals (Sweden)

    C Soldani


    Full Text Available The nucleolus may undergo disassembly either reversibly during mitosis, or irreversibly in apoptosis, thus allowing the redistribution of the nucleolar proteins.We investigated here by immunocytochemistry the fate of three representative proteins, namely phosphorylated c-Myc, fibrillarin and Ki-67, and found that they behave independently in both processes: they relocate in distinct compartments during mitosis, whereas during apoptosis they may either be cleaved (Ki-67 or be extruded into the cytoplasm with a different kinetics and following an ordered, non chaotic program. The separation of these nucleolar proteins which occurs in early apoptotic nuclei continues also in the cytoplasm, and culminates in the final formation of apoptotic blebs containing different nucleolar proteins: this evidence confirms that the apoptotic bodies may be variable in size, content and surface reactivity, and include heterogeneous aggregates of nuclear proteins and/or nucleic acids.

  11. Elevated expression of Ki-67 identifies aggressive prostate cancers but does not distinguish BRCA1 or BRCA2 mutation carriers

    DEFF Research Database (Denmark)

    Mitra, A V; Jameson, C; Barbachano, Y;


    Prostate cancers in men with germline BRCA1 and BRCA2 mutations are more aggressive than morphologically similar cancers in men without these mutations. This study was performed to test the hypothesis that enhanced expression of Ki-67, as a surrogate of cell proliferation, is a characteristic...... feature of prostate cancers occurring in BRCA1 or BRCA2 mutation carriers. The study cohort comprised 20 cases of prostate cancer in mutation carriers and 126 control sporadic prostate cancers. Of the combined sample cohort, 65.7% stained only within malignant tissues while 0.7% stained in both malignant...... a background of BRCA1 or BRCA2 mutations or as sporadic disease. The data suggest that, since elevated Ki-67 does not distinguish prostate cancers occurring in BRCA1 or BRCA2 mutation carriers from sporadic prostatic malignancies, the effects of these genetic mutations are probably independent. While all...

  12. Monoclonal antibodies in animal production; their use in diagnostics and passive immunization.

    NARCIS (Netherlands)

    Booman, P.


    One of the landmarks in immunology was the invention and development of monoclonal antibody-secreting hybridomas by Milstein and his coworkers. The enormous promise of monoclonal antibody technology, which became apparent soon after its discovery, may explain the unusual speed with which monoclonal

  13. Ki-67 Is an Independent Predictor of Metastasis and Cause-Specific Mortality for Prostate Cancer Patients Treated on Radiation Therapy Oncology Group (RTOG) 94-08

    Energy Technology Data Exchange (ETDEWEB)

    Verhoven, Bret [University of Wisconsin Carbone Cancer Center, Madison, Wisconsin (United States); Yan, Yan [RTOG Statistical Center, Philadelphia, Pennsylvania (United States); Ritter, Mark, E-mail: [University of Wisconsin Carbone Cancer Center, Madison, Wisconsin (United States); Khor, Li-Yan [Case Medical Center, Cleveland, Ohio (United States); Hammond, Elizabeth [LDS Hospital, Salt Lake City, Utah (United States); Jones, Christopher [Radiological Associates of Sacramento, Sacramento, California (United States); Amin, Mahul [Cedars-Sinai Medical Center, Los Angeles, California (United States); Bahary, Jean-Paul [Centre Hospitalier de l' Université de Montréal-Notre Dame, Montreal, Ontario (Canada); Zeitzer, Kenneth [Albert Einstein Medical Center, Philadelphia, Pennsylvania (United States); Pollack, Alan [University of Miami Miller School of Medicine, Miami, Florida (United States)


    Purpose: The association of Ki-67 staining index (Ki67-SI) with overall survival (OS), disease-specific mortality (DSM), distant metastasis (DM), and biochemical failure (BF) was examined in men with favorable- to intermediate-risk prostate cancer receiving radiation therapy (RT) alone or with short-term androgen deprivation (ADT) in Radiation Therapy Oncology Group (RTOG) 94-08. Methods and Materials: 468 patients (23.6%) on RTOG 94-08 had sufficient tissue for Ki67-SI analysis. The median follow-up time was 7.9 years. Ki67-SI was determined by immunohistochemistry and quantified manually and by image analysis. Correlative analysis versus clinical outcome was performed using the third quartile (≥Q3) cutpoint. A proportional hazards multivariable analysis (MVA) dichotomized covariates in accordance with trial stratification and randomization criteria. Results: In MVAs adjusted for all treatment covariates, high Ki67-SI (≥Q3) was correlated with increased DSM (hazard ratio [HR] 2.48, P=.03), DM (HR 3.5, P=.002), and BF (HR 3.55, P<.0001). MVA revealed similar Ki67-associated hazard ratios in each separate treatment arm for DSM, DM, and BF; these reached significance only for DM in the RT-alone arm and for BF in both arms. Ki67-SI was not a significant predictor of intraprostatic recurrence assessed by repeated biopsy 2 years after treatment. Patients with a high or low Ki67-SI seemed to experience a similar relative benefit from the addition of ADT to radiation. Conclusions: High Ki67-SI independently predicts for increased DSM, DM, and protocol BF in primarily intermediate-risk prostate cancer patients treated with RT with or without ADT on RTOG 94-08 but does not predict for local recurrence or for increased relative benefit from ADT. This and prior studies lend support for the use of Ki67-SI as a stratification factor in future trials.

  14. Sometimes it takes darkness to see the light: pitfalls in the interpretation of cell proliferation markers (Ki-67 and PCNA). (United States)

    Castilla, Carmen; McDonough, Patrick; Tumer, Gizem; Lambert, Peter C; Lambert, W Clark


    The degree of cell proliferation in a tumor is often associated with metastatic risk and mortality. Proliferating cell nuclear antigen (PCNA) and Ki-67 are proliferation markers that can be used to assess malignant potential in cutaneous lesions and pathological cell proliferation in psoriasis. These markers are elevated during periods of cell proliferation; however, they are also upregulated following UV irradiation. This upregulation may be problematic, as many skin lesions are subject to sun exposure in an everyday setting.

  15. Ki-67 and MCM-2 in Dental Follicle and Odontogenic Cysts: The Effects of Inflammation on Proliferative Markers

    Directory of Open Access Journals (Sweden)

    Nurhan Güler


    Full Text Available The aim of this study was to investigate whether there is any association between inflammation and the expression of markers of cell cycle entry (Ki-67 and MCM-2 in dental follicle (DF of asymptomatic impacted teeth and odontogenic cysts. The study consisted of 70 DFs and 20 odontogenic cysts (radicular cyst (RC, dentigerous cyst (DC and keratocytic odontogenic tumor (KCOT located at posterior mandibular region. Histological findings of inflammation for all specimen and mucous cell prosoplasia, squamous metaplasia, glandular epithelium for all DFs were stained with hematoxyline and eosin, periodic acid schiff, alcian blue, and mucin. Epithelial cell proliferation was determined by using immunohistochemical labeling for Ki-67 and MCM-2. The histologic examinations showed 16% mucous cell prosoplasia, 54% squamous metaplasia, 20% glandular epithelium, 37% inflammation. Inflammation was detected in all RCs and %62 in DF, %43 in DC and KCOT. Positive correlation was found between the inflammation of DF and odontogenic cysts (P<0.01. The mean Ki-67 and MCM-2 expressions were found 9,64±5,99 and 6,34±3,81 in DF, 11,85±9,01 and 13,6±9,94 in odontogenic cysts, respectively. While the mean Ki-67 expressions were statistically significant in DF and KCOT (P<0.01, MCM-2 were significant in RC and KCOT (P<0.01. MCM-2 expresion in RCs were statistically significant than KCOT (P<0.01. The results of this study indicated that the higher MCM-2 expressions in RC than the KCOT might be related to the inflammation and this protein might be more sensitive to inflammation.

  16. Expression of COX-2, PCNA, Ki-67 and p53 in gastrointestinal stromal tumors and its relationship with histopathological parameters

    Institute of Scientific and Technical Information of China (English)

    Derya Gumurdulu; Seyda Erdogan; Fazilet Kayaselcuk; Gulsah Seydaoglu; Cem K Parsak; Orhan Demircan; Ilhan Tuncer


    AIM: To investigate the expression of Cyclooxygenase-2(COX-2), proliferating cell nuclear antigen (PCNA), Ki-67and p53 in gastrointestinal stromal tumors (GISTs) and its relationship with histopathological parameters.METHODS: Twenty-five GISTs were examined by light microscopy and immunohistochemistry. c-kit, CD34,SMA, S-100 protein, COX-2, PCNA, Ki-67 and p53 were detected immunohistochemically and the relationship was evaluated among histopathologic parameters such as mitotic index (MI), tumor grade, tumor size, COX-2,PCNA, Ki-67 and p53.RESULTS: COX-2 protein expression was found in 19 of 25 (76%) of the tumors, and expression was noted in the cytoplasm of the tumor cells. p53 was significantly related to MI and tumor grade but no relationship was found between COX-2, proliferation markers and MI,tumor grade and tumor size.CONCLUSION: COX-2 is expressed in most GISTs and it may play an important role in the proliferation and progression of these tumors or a useful marker to identify GIST. Although immunohistochemical assessment of p53 can be used for distinguishing the risk groups of GISTs, tumor size and mitotic rate should be considered at the same time.

  17. CD147 and Ki-67 overexpression confers poor prognosis in squamous cell carcinoma of oral tongue: A tissue microarray study (United States)

    Yu, Yau-Hua; Morales, Jose; Feng, Lei; Lee, J. Jack; El-Naggar, Adel K.; Vigneswaran, Nadarajah


    Squamous cell carcinoma of the oral tongue (SCCOT) exhibits high risk for recurrence and regional metastasis even after surgical resection. We assessed the clinicopathologic and prognostic significance of a group of functionally related biomarkers. We used a tissue microarray consisting SCCOT from 32 patients for this study. These patients were treated at the UT- M.D. Anderson Cancer Center from 1995 to 2008. Biomarker expression levels were examined by immunohistochemistry and graded semiquantitatively to determine their prognostic significance. CD147 and Tp63 expressions were significantly associated with a higher T-stage and Ki-67 labelling index as well as shorter overall survival (OS). Expression of Tp63 associated positively with poorly-differentiated histology. There was significant association of Tp63 with the expression levels of CD147 and Glut-1. Glut-1 overexpression was marginally associated with a higher T-stage. There was no prognostic significance of CD44v6 expression in SCCOT. SCCOT with CD147 overexpression in combination with high Ki-67 labelling index had poor OS. CD147 and Ki-67 overexpression is associated with aggressive disease with poor prognosis in SCCOT. PMID:25747176

  18. p53 is not related to Ki-67 immunostaining in the epithelial and mesenchymal components of female genital tract carcinosarcomas. (United States)

    Bałon, Beata; Kaznowska, Ewa; Ignatov, Atanas; Steć, Anna; Semczuk-Sikora, Anna; Schneider-Stock, Regine; Jóźwik, Maciej; Sulkowski, Stanisław; Cybulski, Marek; Szumiło, Justyna; Semczuk, Andrzej


    Carcinosarcomas (CSs) are composed of two separate histological components and are rare neoplasms of the female genital tract. Therefore, CS pathogenesis has not yet been fully elucidated. In the present study, immunohistochemical techniques were used to determine the role of p53 and Ki-67 overexpression in female genital tract CSs. The study group was comprised of 36 patients with CSs originating from the uterus (n=31), cervix (n=3) and ovary (n=2), as well as 3 metastatic tissues. p53 was overexpressed in the epithelial component of 23 out of 36 (64%) tumors, and in the mesenchymal component of 20 out of 36 (56%) tumors. In both CS components, there was a significant correlation between p53 overexpression and patient age and ovarian metastases. Ki-67 overexpression was detected in the epithelial component in 15 out of 36 (42%) cases, and in the mesenchymal component in 13 out of 36 (36%) neoplasms. There was a significant correlation of p53 overexpression between the carcinomatous and sarcomatous components (R=0.884, Ptumor components. In conclusion, based on immunohistochemical results, p53 was overexpressed in more than half of the female genital tract CSs included in the present study, either at the epithelial or mesenchymal component. The correlation between p53 or Ki-67 overexpression in both tumor components supports the combination theory of histogenesis in the majority of these tumors.

  19. Ofatumumab: a novel monoclonal anti-CD20 antibody

    Directory of Open Access Journals (Sweden)

    Thomas S Lin


    Full Text Available Thomas S LinGlaxoSmithKline Oncology R&D, Collegeville, PA, USAAbstract: Ofatumumab, a novel humanized monoclonal anti-CD20 antibody, was recently approved by the FDA for the treatment of fludarabine and alemtuzumab refractory chronic lymphocytic leukemia (CLL. Ofatumumab effectively induces complement-dependent cytotoxicity (CDC in vitro, and recent studies demonstrated that ofatumumab also effectively mediates antibody-dependent cellular cytotoxicity (ADCC. Pharmacokinetic studies indicated that increased exposure to the antibody correlated with improved clinical outcome in CLL. Thus, pharmacogenomics may be important in identifying which patients are more likely to respond to ofatumumab therapy, although such studies have not yet been performed. Patients with the high-affinity FCGR3a 158 V/V polymorphism may be more likely to respond to therapy, if ADCC is the primary in vivo mechanism of action of ofatumumab. Patients with increased expression of the complement defense proteins CD55 and CD59 may be less likely to respond if ofatumumab works in vivo primarily via CDC. Patients with increased metabolism and clearance of ofatumumab may have lower exposure and be less likely to respond clinically. Thus, pharmacogenomics may determine the responsiveness of patients to ofatumumab therapy.Keywords: monoclonal antibody, CD20, CLL, NHL, lymphoma

  20. Monoclonal Antibody Production against Human Spermatozoal Surface Antigens

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    M Jedi-Tehrani


    Full Text Available Introduction: As monoclonal antibodies are potential tools for characterization of soluble or cellular surface antigens, use of these proteins has always been considered in infertility and reproduction research. Therefore, in this study, monoclonal antibodies against human sperm surface antigens were produced. Material and Methods: To produce specific clones against human sperm surface antigens, proteins were extracted using solubilization methods. Balb/c mice were immunized intraperitoneally with the proteins using complete Freund’s adjuvant in the first injection and incomplete Adjuvant in the following booster injections. Hybridoma cells producing ASA were cloned by limiting dilution. Results: Five stable ASA producing hybridoma clones were achieved and their antibody isotypes were determined by ELISA. All the isotypes were of IgG class. Their cross reactivity with rat and mice spermatozoa was examined but they did not have any cross reactivity. Conclusion: The produced antibodies can be used in further studies to characterize and evaluate each of the antigens present on human sperm surface and determining their role in fertilization.

  1. Prediction and Reduction of the Aggregation of Monoclonal Antibodies. (United States)

    van der Kant, Rob; Karow-Zwick, Anne R; Van Durme, Joost; Blech, Michaela; Gallardo, Rodrigo; Seeliger, Daniel; Aßfalg, Kerstin; Baatsen, Pieter; Compernolle, Griet; Gils, Ann; Studts, Joey M; Schulz, Patrick; Garidel, Patrick; Schymkowitz, Joost; Rousseau, Frederic


    Protein aggregation remains a major area of focus in the production of monoclonal antibodies. Improving the intrinsic properties of antibodies can improve manufacturability, attrition rates, safety, formulation, titers, immunogenicity, and solubility. Here, we explore the potential of predicting and reducing the aggregation propensity of monoclonal antibodies, based on the identification of aggregation-prone regions and their contribution to the thermodynamic stability of the protein. Although aggregation-prone regions are thought to occur in the antigen binding region to drive hydrophobic binding with antigen, we were able to rationally design variants that display a marked decrease in aggregation propensity while retaining antigen binding through the introduction of artificial aggregation gatekeeper residues. The reduction in aggregation propensity was accompanied by an increase in expression titer, showing that reducing protein aggregation is beneficial throughout the development process. The data presented show that this approach can significantly reduce liabilities in novel therapeutic antibodies and proteins, leading to a more efficient path to clinical studies. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  2. Efficient generation of human IgA monoclonal antibodies. (United States)

    Lorin, Valérie; Mouquet, Hugo


    Immunoglobulin A (IgA) is the most abundant antibody isotype produced in humans. IgA antibodies primarily ensure immune protection of mucosal surfaces against invading pathogens, but also circulate and are present in large quantities in blood. IgAs are heterogeneous at a molecular level, with two IgA subtypes and the capacity to form multimers by interacting with the joining (J) chain. Here, we have developed an efficient strategy to rapidly generate human IgA1 and IgA2 monoclonal antibodies in their monomeric and dimeric forms. Recombinant monomeric and dimeric IgA1/IgA2 counterparts of a prototypical IgG1 monoclonal antibody, 10-1074, targeting the HIV-1 envelope protein, were produced in large amounts after expression cloning and transient transfection of 293-F cells. 10-1074 IgAs were FPLC-purified using a novel affinity-based resin engrafted with anti-IgA chimeric Fabs, followed by a monomers/multimers separation using size exclusion-based FPLC. ELISA binding experiments confirmed that the artificial IgA class switching of 10-1074 did not alter its antigen recognition. In summary, our technical approach allows the very efficient production of various forms of purified recombinant human IgA molecules, which are precious tools in dissecting IgA B-cell responses in physiological and pathophysiological conditions, and studying the biology, function and therapeutic potential of IgAs.

  3. Radiolabelled peptides and monoclonal antibodies for therapy decision making in inflammatory diseases

    NARCIS (Netherlands)

    Malviya, G.; Signore, A.; Lagana, B.; Dierckx, R. A.


    Radiolabelled peptides and monoclonal antibodies are an emerging class of radiopharmaceuticals for imaging inflammation with clinical implications for several chronic inflammatory disorders for diagnosis, therapy decision making and follow up. In the last decades, a number of novel monoclonal antibo

  4. Interval sentinel lymph nodes in melanoma: a digital pathology analysis of Ki67 expression and microvascular density. (United States)

    Marinaccio, Christian; Giudice, Giuseppe; Nacchiero, Eleonora; Robusto, Fabio; Opinto, Giuseppina; Lastilla, Gaetano; Maiorano, Eugenio; Ribatti, Domenico


    The presence of interval sentinel lymph nodes in melanoma is documented in several studies, but controversies still exist about the management of these lymph nodes. In this study, an immunohistochemical evaluation of tumor cell proliferation and neo-angiogenesis has been performed with the aim of establishing a correlation between these two parameters between positive and negative interval sentinel lymph nodes. This retrospective study reviewed data of 23 patients diagnosed with melanoma. Bioptic specimens of interval sentinel lymph node were retrieved, and immunohistochemical reactions on tissue sections were performed using Ki67 as a marker of proliferation and CD31 as a blood vessel marker for the study of angiogenesis. The entire stained tissue sections for each case were digitized using Aperio Scanscope Cs whole-slide scanning platform and stored as high-resolution images. Image analysis was carried out on three selected fields of equal area using IHC Nuclear and Microvessel analysis algorithms to determine positive Ki67 nuclei and vessel number. Patients were divided into positive and negative interval sentinel lymph node groups, and the positive interval sentinel lymph node group was further divided into interval positive with micrometastasis and interval positive with macrometastasis subgroups. The analysis revealed a significant difference between positive and negative interval sentinel lymph nodes in the percentage of Ki67-positive nuclei and mean vessel number suggestive of an increased cellular proliferation and angiogenesis in positive interval sentinel lymph nodes. Further analysis in the interval positive lymph node group showed a significant difference between micro- and macrometastasis subgroups in the percentage of Ki67-positive nuclei and mean vessel number. Percentage of Ki67-positive nuclei was increased in the macrometastasis subgroup, while mean vessel number was increased in the micrometastasis subgroup. The results of this study suggest

  5. Studies on Purification of Methamidophos Monoclonal Antibodies and Comoarative Immunoactivity of Purified Antibodies

    Institute of Scientific and Technical Information of China (English)



    Objective To purify Methamidophos (Met) monoclonal antibodies with two methods andcompare immune activity of purified antibodies. Method Caprylic acid ammonium sulphateprecipition (CAASP) method and Sepharose protein-A (SPA) affinity chromatography method wereused to purify Met monoclonal antibodies, UV spectrum scanning was used to determine proteincontent and recovery of purified antibodies, sodium dodecylsulphate polyacrylamide gelelectrophoresis (SDS-PAGE) was used to analyze the purity of purified antibodies, and enzyme-linkedimmunosorbent assay (ELISA) was used to determine immune activity of purified antibodies.Results Antibody protein content and recovery rate with CAASP method were 7.62 mg/mL and8.05% respectively, antibody protein content and recovery rate with SPA method were 6.45 mg/mLand 5.52% respectively. Purity of antibodies purified by SPA method was higher than that by CAASPmethod. The half-maximal inhibition concentration (IC50) of antibodies purified by SPA to Met was181.26 μg/mL, and the linear working range and the limit of quantification (LOD) were 2.43-3896.01μg/mL and 1.03 μg/mL, respectively. The IC50 of antibodies purified by CAASP to Met was 352.82μg/mL, and the linear working range and LOD were 10.91-11412.29 ug/mL and 3.42 μg/mL,respectively. Conclusion Antibodies purified by SPA method are better than those by CAASPmethod, and Met monoclonal antibodies purified by SPA method can be used to prepare gold-labelledtesting paper for analyzing Met residue in vegetable and drink water.

  6. Modulation of desmin intermediate filament assembly by a monoclonal antibody (United States)


    We have used a monoclonal antibody against desmin to examine the assembly of intermediate filaments (IF) from their building blocks, the tetrameric protofilaments. The antibody, designated D76, does not cross react with any other IF proteins (Danto, S.I., and D.A. Fischman. 1984. J. Cell Biol. 98:2179-2191). It binds to a region amino-terminal to cys- 324 of avian desmin that is resistant to chymotrypsin and trypsin digestion, and in the electron microscope appears to bind to the ends of tetrameric protofilaments. In combination, these findings suggest that the epitope of the antibody resides at the amino-terminal end of the alpha-helical rod domain. Preincubation of desmin protofilaments with an excess of D76 antibodies blocks their subsequent assembly into IF. In the presence of sub-stoichiometric amounts of antibodies, IF are assembled from protofilaments but they are morphologically aberrant in that (a) they are capped by IgG molecules at one or both ends; (b) they are unraveled to varying degree, revealing a characteristic right- handed helical arrangement of sub-filamentous strands of different diameters. The antibody binds only to the ends but not along the length of desmin IF. The most straightforward explanation for this is that the epitope resides in a part of the desmin molecule that becomes buried within the core of the filament upon polymerization and is therefore inaccessible to the antibody. PMID:2450097

  7. Monitoring monoclonal antibody delivery in oncology: the example of bevacizumab.

    Directory of Open Access Journals (Sweden)

    Guillaume Nugue

    Full Text Available Developing therapeutic monoclonal antibodies paves the way for new strategies in oncology using targeted therapy which should improve specificity. However, due to a lack of biomarkers, a personalized therapy scheme cannot always be applied with monoclonal antibodies. As a consequence, the efficacy or side effects associated with this type of treatment often appear to be sporadic. Bevacizumab is a therapeutic monoclonal antibody targeting Vascular Endothelial Growth Factor (VEGF. It is used to limit tumor vascularization. No prognosis or response biomarker is associated with this antibody, we therefore assessed whether the administration protocol could be a possible cause of heterogeneous responses (or variable efficacy. To do this, we developed a bevacizumab assay with a broad sensitivity range to measure blood bevacizumab concentrations. We then analyzed bevacizumab concentrations in 17 patients throughout the first quarter of treatment. In line with previously published data, average blood concentrations were 88+/-27 mg/L following the first dose administered, and 213+/-105 mg/L after the last (6(th dose administered. However, the individual values were scattered, with a mean 4-fold difference between the lowest and the highest concentration for each dose administered. We demonstrated that the bevacizumab administration schedule results in a high inter-individual variability in terms of blood concentrations. Comparison of assay data with clinical data indicates that blood concentrations above the median are associated with side effects, whereas values below the median favor inefficacy. In conclusion, bevacizumab-based therapy could benefit from a personalized administration schedule including follow-up and adjustment of circulating bevacizumab concentrations.

  8. Estudo citofotométrico da expressão dos marcadores tumorais Caspase-3 e Ki-67 no adenocarcinoma gástrico Cytophotometric study of the expression of tumoral markers Caspase-3 and Ki-67 in gastric adenocarcinoma

    Directory of Open Access Journals (Sweden)

    Pedro Manuel Gonzales Cuellar


    Full Text Available RACIONAL: A carcinogênese gástrica é processo complexo e depende de fatores genéticos, ambientais e infecciosos. Nos últimos anos, houve grandes avanços nos campos da genética e da biologia molecular, sobre o desenvolvimento dos tumores. Os marcadores tumorais são substâncias ausentes nos tecidos normais e que podem ser identificadas em tecidos com câncer. Através de procedimentos imunoistoquímicos eles podem ser estudados. OBJETIVOS: Descrever a expressão citofotométrica do marcador tumoral Ki-67 analisando a densidade óptica e o índice de marcagem no adenocarcinoma de estômago. Descrever a expressão citofotométrica do marcador tumoral Caspase-3 analisando a densidade óptica e o índice de marcagem no adenocarcinoma de estômago. Comparar o índice de marcagem e densidade óptica dos marcadores tumorais Ki-67 e Caspase-3 no adenocarcinoma de estômago. MÉTODO: Foram selecionados, inicialmente, 58 blocos com espécime de adenocarcinoma gástrico coletados nos Serviços de Anátomo-Patologia do Hospital do Gama - Brasília (DF e Hospital Dom Orione - Araguaina (TO, e analizados no Laboratório de Citologia e Histopalogia Ltda - CITOLAB, Curitiba (PR. Foram aproveitados 31 blocos para o estudo histológico e imunoistoquímico realizado pelo sistema de análise computarizado SAMBA 4000. RESULTADOS: Das 31 lâminas estudas, 15 (48% foram marcadas pelo marcador Ki-67, 22 (71% foram marcadas pelo marcador Caspase-3 e 14 (45% marcaram com os dois marcadores. CONCLUSÕES: A expressão citofométrica do marcador Ki-67 foi observada em 15 lâminas da amostra estudada e apresentaram média do índice de marcagem de 36,85%, enquanto a densidade óptica apresentou média de 29,33 pixels. A expressão citofotométrica do marcador Caspase-3 foi observada em 22 lâminas da amostra estudada e apresentaram média do índice de marcagem de 87,71% e 60,74 pixels de média para a densidade óptica. Na comparação do índice de marcagem dos

  9. Correlation between semi-quantitative {sup 18}F-FDG PET/CT parameters and Ki-67 expression in small cell lung cancer

    Energy Technology Data Exchange (ETDEWEB)

    Park, So Yeon; Lee, Eun Sub; Eo, Jae Seon [Dept. of Nuclear Medicine, Korea University Guro Hospital, Seoul (Korea, Republic of); Rhee, Seung Hong; Cho, Jae Hyuk; Choi, Sun Ju; Pahk, Kisso; Choe, Jae Gol; Kim, Sung Geun [Dept. of Nuclear Medicine, Korea University Anam Hospital, Seoul (Korea, Republic of); Lee, Si Nae [Dept. of Nuclear Medicine, G Sam Hospital, Gunpo (Korea, Republic of)


    The aim of this study was to evaluate the relationship between semiquantitative parameters on {sup 18}F-FDG PET/CT including maximum standardized uptake value (SUV{sub max}), mean standardized uptake value (SUV{sub mean}), metabolic tumor volume (MTV), and total lesion glycolysis (TLG) and the expression level of Ki-67 in small-cell lung cancer (SCLC). Ninety-four consecutive patients with SCLC were enrolled in this study. They underwent {sup 18}F-FDG PET/CT for initial evaluation of SCLC, and we measured SUV{sub max}, {sub avg}SUV{sub mean}, MTV{sub sum}, and TLGtotal on {sup 18}F-FDG PET/CT images. The protein expression of Ki-67 was examined by immunohistochemical staining. Significant correlations were found between the MTVsum and Ki-67 labeling index (r=0.254, p=0.014) and the TLGtotal and Ki-67 labeling index (r=0.239, p=0.020). No correlation was found between the SUVmax and Ki-67 labeling index (r=0.116, p=0.264) and the {sub avg}SUV{sub mean} and Ki-67 labeling index (r=0.031, p=0.770). Dividing the Ki-67 expression level into three categories, it was suggested that increasing Ki-67 expression level caused a stepwise increase in the MTV{sub sum} and TLGtotal. (p=0.028 and 0.039, respectively), but not the SUV{sub max} and {sub avg}SUV{sub mean} (p=0.526 and 0.729, respectively). In conclusion, the volume-based parameters of {sup 18}F-FDG PET/CT correlate with immunohistochemical staining of Ki-67 in SCLC. Measurement of the MTV{sub sum} and TLGtotal by {sup 18}F-FDG PET/CT might be a simple, noninvasive, and useful method to determine the proliferative potential of cancer cells.

  10. Membrane adsorbers as purification tools for monoclonal antibody purification. (United States)

    Boi, Cristiana


    Downstream purification processes for monoclonal antibody production typically involve multiple steps; some of them are conventionally performed by bead-based column chromatography. Affinity chromatography with Protein A is the most selective method for protein purification and is conventionally used for the initial capturing step to facilitate rapid volume reduction as well as separation of the antibody. However, conventional affinity chromatography has some limitations that are inherent with the method, it exhibits slow intraparticle diffusion and high pressure drop within the column. Membrane-based separation processes can be used in order to overcome these mass transfer limitations. The ligand is immobilized in the membrane pores and the convective flow brings the solute molecules very close to the ligand and hence minimizes the diffusional limitations associated with the beads. Nonetheless, the adoption of this technology has been slow because membrane chromatography has been limited by a lower binding capacity than that of conventional columns, even though the high flux advantages provided by membrane adsorbers would lead to higher productivity. This review considers the use of membrane adsorbers as an alternative technology for capture and polishing steps for the purification of monoclonal antibodies. Promising industrial applications as well as new trends in research will be addressed.

  11. Monoclonal Antibody Therapy and Renal Transplantation: Focus on Adverse Effects

    Directory of Open Access Journals (Sweden)

    Gianluigi Zaza


    Full Text Available A series of monoclonal antibodies (mAbs are commonly utilized in renal transplantation as induction therapy (a period of intense immunosuppression immediately before and following the implant of the allograft, to treat steroid-resistant acute rejections, to decrease the incidence and mitigate effects of delayed graft function, and to allow immunosuppressive minimization. Additionally, in the last few years, their use has been proposed for the treatment of chronic antibody-mediated rejection, a major cause of late renal allograft loss. Although the exact mechanism of immunosuppression and allograft tolerance with any of the currently used induction agents is not completely defined, the majority of these medications are targeted against specific CD proteins on the T or B cells surface (e.g., CD3, CD25, CD52. Moreover, some of them have different mechanisms of action. In particular, eculizumab, interrupting the complement pathway, is a new promising treatment tool for acute graft complications and for post-transplant hemolytic uremic syndrome. While it is clear their utility in renal transplantation, it is also unquestionable that by using these highly potent immunosuppressive agents, the body loses much of its innate ability to mount an adequate immune response, thereby increasing the risk of severe adverse effects (e.g., infections, malignancies, haematological complications. Therefore, it is extremely important for clinicians involved in renal transplantation to know the potential side effects of monoclonal antibodies in order to plan a correct therapeutic strategy minimizing/avoiding the onset and development of severe clinical complications.

  12. Monoclonal anti-elastin antibody labelled with technetium-99m

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Marcia B.N. de; Silva, Claudia R. da; Araujo, Adriano C. de; Bernardo Filho, Mario [Universidade do Estado, Rio de Janeiro, RJ (Brazil). Instituto de Biologia Roberto Alcantara Gomes. Lab. de Radiofarmacia; Porto, Luis Cristovao M.S.; Gutfilen, Bianca [Universidade do Estado, Rio de Janeiro, RJ (Brazil). Instituto de Biologia Roberto Alcantara Gomes; Souza, J.E.Q. [Instituto Nacional do Cancer, Rio de Janeiro, RJ (Brazil). Centro de Pesquisa Basica; Frier, Malcolm [University Hospital, Nottingham (United Kingdom). Dept. of Medical Physics


    Technetium-99m ({sup 99m} Tc) is widely employed in nuclear medicine due to its desirable physical, chemical and biological properties. Moreover, it is easily available and normally is inexpensive. A reducing agent is necessary to label cells and molecules with {sup 99m} Tc and stannous chloride (Sn C L{sub 2}) is usually employed. Elastin is the functional protein component of the elastic fiber and it is related with some diseases such as arteriosclerosis, pulmonary emphysema and others. The present study refers to the preparation of the {sup 99m} Tc labeled monoclonal anti-elastin antibody. The monoclonal antibody was incubated with an excess of 2-iminothiolane. The free thiol groups created, were capable of binding with the reduced technetium. Labeling was an exchange reaction with {sup 99m} Tc-glucoheptonate. The labeled preparation was left at 4 deg C for one hour. Then, it was passed through a Sephadex G50 column. Various fractions were collected and counted. A peak corresponding to the radiolabeled antibody was obtained. Stability studies of the labelled anti-elastin were performed at 0,3 6, 24 hours, at both 4 deg C or room temperature. The biodistribution pattern of the {sup 99m} Tc-anti-elastin was studied in healthy male Swiss mice. The immunoreactivity was also determined. An useful labeled-anti-elastin was obtained to future immunoscintigraphic investigations. (author) 4 refs., 7 figs., 6 tabs.

  13. Characterization of oxidative carbonylation on recombinant monoclonal antibodies. (United States)

    Yang, Yi; Stella, Cinzia; Wang, Weiru; Schöneich, Christian; Gennaro, Lynn


    In the biotechnology industry, oxidative carbonylation as a post-translational modification of protein pharmaceuticals has not been studied in detail. Using Quality by Design (QbD) principles, understanding the impact of oxidative carbonylation on product quality of protein pharmaceuticals, particularly from a site-specific perspective, is critical. However, comprehensive identification of carbonylation sites has so far remained a very difficult analytical challenge for the industry. In this paper, we report for the first time the identification of specific carbonylation sites on recombinant monoclonal antibodies with a new analytical approach via derivatization with Girard's Reagent T (GRT) and subsequent peptide mapping with high-resolution mass spectrometry. Enhanced ionization efficiency and high quality MS(2) data resulted from GRT derivatization were observed as key benefits of this approach, which enabled direct identification of carbonylation sites without any fractionation or affinity enrichment steps. A simple data filtering process was also incorporated to significantly reduce false positive assignments. Sensitivity and efficiency of this approach were demonstrated by identification of carbonylation sites on both unstressed and oxidized antibody bulk drug substances. The applicability of this approach was further demonstrated by identification of 14 common carbonylation sites on three highly similar IgG1s. Our approach represents a significant improvement to the existing analytical methodologies and facilitates extended characterization of oxidative carbonylation on recombinant monoclonal antibodies and potentially other protein pharmaceuticals in the biotechnology industry.

  14. Improved detection of Pneumocystis carinii by an immunofluorescence technique using monoclonal antibodies

    DEFF Research Database (Denmark)

    Orholm, M; Holten-Andersen, W; Lundgren, Jens Dilling


    To assess whether a recently developed indirect immunofluorescent stain using monoclonal antibodies was more sensitive in detecting Pneumocystis carinii than the combination of Giemsa and methenamine silver nitrate stains which has routinely been used in the laboratory, 88 lavage fluid specimens...... and 34 induced sputum specimens were examined. All specimens were stained by five techniques: immunofluorescence using a combination of three monoclonal antibodies (from the National Institutes of Health, USA), immunofluorescence using a single monoclonal antibody (from Dakopatts), Giemsa, methenamine...

  15. High-efficiency screening of monoclonal antibodies for membrane protein crystallography.

    Directory of Open Access Journals (Sweden)

    Hyun-Ho Lim

    Full Text Available Determination of crystal structures of membrane proteins is often limited by difficulties obtaining crystals diffracting to high resolution. Co-crystallization with Fab fragments of monoclonal antibodies has been reported to improve diffraction of membrane proteins crystals. However, it is not simple to generate useful monoclonal antibodies for membrane protein crystallography. In this report, we present an optimized process for efficient screening from immunization to final validation of monoclonal antibody for membrane protein crystallography.

  16. Expressão citofotométrica dos marcadores tumorais Ki-67 e CD34 no adenocarcinoma de estômago Citophotometric expression of tumoral markers: Ki67 and CD34 in stomach adenocarcinoma

    Directory of Open Access Journals (Sweden)

    José Augusto Menezes Freitas de Campos


    Full Text Available RACIONAL: O câncer de estômago ocupa o terceiro lugar das neoplasias malignas em todo o mundo e é a segunda causa de morte por câncer. No entanto, sua incidência está em declínio nos últimos anos, principalmente em paises como Estados Unidos, Inglaterra e Austrália, assim como onde a sua incidência sempre foi alta - Japão, China, Coréia e Leste da Ásia -, com estimativa de 780 casos anuais por 100 mil habitantes. OBEJTIVOS: Descrição da expressão citofotométrica dos marcadores Ki-67 e CD34 e comparação entre a expressão deles no adenocarcinoma de estômago. MÉTODOS: Foram selecionados inicialmente 60 blocos com espécime de adenocarcinoma gástrico coletados nos serviços de patologia do Hospital do Gama - Brasília, DF e Hospital Dom Orione - Araguaina,TO. A imunoistoquímica com os marcadores Ki-67 e CD34 foi feita no Laboratório de Citologia e Histopalogia Ltda - CITOLAB, Curitiba, PR. Foram aproveitados do total 35 blocos para o estudo histológico e imunoistoquímico que foram analisados pelo sistema de citometria de imagem computarizado SAMBA 4000 no Instituto de Pesquisas Médicas do Hospital Universitário Evangélico de Curitiba. Foram estudados o índice de marcagem e a densidade óptica para ambos os marcadores analisados separadamente e depois comparados. RESULTADOS: Das 35 lâminas, 15 (42,85% marcaram o Ki-67; 26 (74,28% o CD34 e 12 lâminas (34,28% marcaram com os dois marcadores. Quando se comparou o índice de marcagem entre Ki-67 e CD34, observou-se que existe diferença significativa (PBACKGROUND: Stomach cancer is rated as the third malignant neoplasm in the world, being the second most frequent cause of death. However, its incidence has been declining in the last few years in countries like the United States, England and Australia, and in countries where its incidence has always been high - Japan, China, Korea and in the Eastern part of Asia, which has an estimated 780 cases per year per 100

  17. Expresión de bcl-2, ki-67 y caspasa-3 en lesiones cancerosas de la mucosa oral: Resultados preliminares Blc-2, ki-67 and caspasa-3 expression in oral cancer lesions: Preliminary results

    Directory of Open Access Journals (Sweden)

    V García García


    Full Text Available El cáncer oral representa del 2 al 4% de todos los cánceres diagnosticados, siendo el carcinoma epidermoide el tumor más común encontrado a nivel histológico. En varios estudios se ha constatado que el uso de la clasificación TNM del cáncer oral no resulta de utilidad a la hora de establecer el pronóstico tumoral. Nos encontramos en la era de los marcadores tumorales que actualmente pueden ser detectados y medidos con las más modernas técnicas de inmunohistoquímica. Material y método: aplicación de la técnica inmunohistoquímica peroxidasa- antiperoxidasa para la detección de las moléculas bcl-2, caspasa-3 y Ki-67 en lesiones cancerosas. Resultados preliminares y discusión: la detección molecular del marcador antiapoptótico bcl-2, el marcador proapoptótico caspasa 3 y el marcador de proliferación celular Ki-67 nos informará sobre la situación más o menos grave del paciente respecto de su proceso canceroso.Oral cancer represents 2%-4% of all types of cancer that are diagnosed and epidermal carcinoma is the most common tumour found in oral cavity. Some experimental studies have shown that TNM classification is not useful when you want to know tumour prognosis. Now we are in the modern inmunohistochemical period with new techniques for cancer diagnose. Material and method: use the peroxidase-antiperoxidase inmunohistochemical technique to detect bcl-2, caspasa-3 and Ki-67 molecules in cancerous lesions. Preliminary results and discussion: molecular detection of the antiapoptotic bcl-2 molecule, the proapoptotic caspasa-3 molecule and the cellular proliferation indicator Ki-67 will show us about the gravity of the patient situation.

  18. Alemtuzumab and Natalizumab: The Monoclonal Antibody Story Continues

    Directory of Open Access Journals (Sweden)

    BL Johnston


    Full Text Available In the July/August 2006 issue of this journal, the infectious complications associated with the use of infliximab, etanercept and adalimumab were reviewed (1. These represent only three of the many monoclonal antibodies either licensed or in clinical trials for therapeutic use in cancer and autoimmune disease or to prevent rejection in both solid organ and hematopoietic stem cell transplantation. While most of these agents have not been associated with increased infection rates, alemtuzumab and natalizumab have gained particular attention related to either the frequency or type of infection seen in some individuals who have received them.

  19. Rapid analysis of small samples containing forskolin using monoclonal antibodies. (United States)

    Yanagihara, H; Sakata, R; Shoyama, Y; Murakami, H


    The effective range of the competitive ELISA test for detection of forskolin content in clonally propagated plant organs of Coleus forskohlii using monoclonal antibodies extends from 5ng to 5 micrograms. A correlation between the forskolin accumulation and the growth rate was investigated using the clonally propagated shoots. An increase of forskolin content was noted, beginning at week 6. Flowers, rachises, leaves, stems, tuberous roots, and roots were analyzed. Tuberous roots and the stem base contained higher amounts of forskolin than other organs. The forskolin content in the stem decreased gradually towards the top of the shoot.

  20. Fluorescence polarization immunoassay for salinomycin based on monoclonal antibodies

    Institute of Scientific and Technical Information of China (English)


    A fluorescence polarization immunoassay(FPIA) for the determination of salinomycin(SAL) was developed by using anti-SAL monoclonal antibodies(mAb).Fluorescein labeled SAL(tracer) was synthesized by the N-hydroxysuccinimide active ester method and purified using thin layer chromatography(TLC).The developed FPIA for SAL had a dynamic range from 0.60 to 2193 ng/mL with an IC50 value of 33.2 ng/mL and a detection limit(LOD) of 0.08 ng/mL.No significant cross-reactivities were observed with other drugs but 67.6% with narasin.

  1. Monoclonal antibody therapy in the treatment of Reye's syndrome. (United States)

    Treon, S P; Broitman, S A


    A role for lipopolysaccharides (endotoxins, LPS) in 7 the pathogenesis of Reye's syndrome (RS) has previously been suggested. Impairment of hepatic LPS clearance can lead to systemic endotoxemia as previous studies by this and other laboratories have suggested for several hepatic disorders including RS. Systemic LPS may mediate many of the clinical findings associated with RS by eliciting monokines such as tumor necrosis factor-alpha, interleukin-1, interleukin-6, and interleukin-8. Monoclonal antibody therapy directed at LPS, and monokines may represent a novel approach to the treatment of RS.

  2. Large-scale production of monoclonal antibodies in suspension culture. (United States)

    Backer, M P; Metzger, L S; Slaber, P L; Nevitt, K L; Boder, G B


    Monoclonal antibodies are being manufactured for clinical trials in suspension culture at the 1300-L scale. Suspension culture offers some advantages relative to high-density mammalian cell culture methods; in particular, the ability to closely monitor the behavior of cells in a homogeneous environment. Computer control and on-line mass spectrography of exit gases provide instantaneous information about the culture metabolic activity. Air sparging and agitation by marine impeller provide aeration sufficient to maintain a constant dissolved oxygen tension at cell concentrations up to 5.0 x 10(6) cells/mL without causing apparent cell damage.

  3. Monoclonal Antibodies as Prophylactic and Therapeutic Agents Against Chikungunya Virus. (United States)

    Clayton, April M


    Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that is responsible for considerable epidemics worldwide and recently emerged in the Americas in 2013. CHIKV may cause long-lasting arthralgia after acute infection. With currently no licensed vaccines or antivirals, the design of effective therapies to prevent or treat CHIKV infection is of utmost importance and will be facilitated by increased understanding of the dynamics of chikungunya. In this article, monoclonal antibodies against CHIKV as viable prophylactic and therapeutic agents will be discussed. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail

  4. Production and Characterization of Monoclonal Antibodies of Shrimp White Spot Syndrome Virus Envelope Protein VP28

    Institute of Scientific and Technical Information of China (English)

    Wan-gang GU; Jun-fa YUAN; Ge-lin XU; Li-juan LI; Ni LIU; Cong ZHANG; Jian-hong ZHANG; Zheng-li SHI


    BALB/c mice were immunized with purified White spot syndrome virus (WSSV).Six monoclonal antibody cell lines were selected by ELISA with VP28 protein expressed in E.coll in vitro neutralization experiments showed that 4 of them could inhibit the virus infection in crayfish.Westernblot suggested that all these monoclonal antibodies were against the conformational structure of VP28.The monoclonal antibody 7B4 was labeled with colloidal gold particles and used to locate the VP28 on virus envelope by immunogold labeling.These monoclonal antibodies could be used to develop immunological diagnosis methods for WSSV infection.

  5. Monoclonal antibodies to intermediate filament proteins of human cells: unique and cross-reacting antibodies. (United States)

    Gown, A M; Vogel, A M


    Monoclonal antibodies were generated against the intermediate filament proteins of different human cells. The reactivity of these antibodies with the different classes of intermediate filament proteins was determined by indirect immunofluorescence on cultured cells, immunologic indentification on SDS polyacrylamide gels ("wester blot" experiments), and immunoperoxidase assays on intact tissues. The following four antibodies are described: (a) an antivimentin antibody generated against human fibroblast cytoskeleton; (b), (c) two antibodies that recognize a 54-kdalton protein in human hepatocellular carcinoma cells; and (d) an antikeratin antibody made to stratum corneum that recognizes proteins of molecular weight 66 kdaltons and 57 kdaltons. The antivimentin antibody reacts with vimentin (58 kdaltons), glial fibrillary acidic protein (GFAP), and keratins from stratum corneum, but does not recognize hepatoma intermediate filaments. In immunofluorescence assays, the antibody reacts with mesenchymal cells and cultured epithelial cells that express vimentin. This antibody decorates the media of blood vessels in tissue sections. One antihepatoma filament antibody reacts only with the 54 kdalton protein of these cells and, in immunofluorescence and immunoperoxidase assays, only recognizes epithelial cells. It reacts with almost all nonsquamous epithelium. The other antihepatoma filament antibody is much less selective, reacting with vimentin, GFAP, and keratin from stratum corneum. This antibody decorates intermediate filaments of both mesenchymal and epithelial cells. The antikeratin antibody recognizes 66-kdalton and 57-kdalton proteins in extracts of stratum corneum and also identifies proteins of similar molecular weights in all cells tested. However, by immunofluorescence, this antibody decorates only the intermediate filaments of epidermoid carcinoma cells. When assayed on tissue sections, the antibody reacts with squamous epithelium and some, but not all

  6. Monoclonal antibody-based candidate therapeutics against HIV type 1. (United States)

    Chen, Weizao; Dimitrov, Dimiter S


    Treatment of HIV-1 infection has been highly successful with small molecule drugs. However, resistance still develops. In addition, long-term use can lead to toxicity with unpredictable effects on health. Finally, current drugs do not lead to HIV-1 eradication. The presence of the virus leads to chronic inflammation, which can result in increased morbidity and mortality after prolonged periods of infection. Monoclonal antibodies (mAbs) have been highly successful during the past two decades for therapy of many diseases, primarily cancers and immune disorders. They are relatively safe, especially human mAbs that have evolved in humans at high concentrations to fight diseases and long-term use may not lead to toxicities. Several broadly neutralizing mAbs (bnmAbs) against HIV-1 can protect animals but are not effective when used for therapy of an established infection. We have hypothesized that HIV-1 has evolved strategies to effectively escape neutralization by full-size antibodies in natural infections but not by smaller antibody fragments. Therefore, a promising direction of research is to discover and exploit antibody fragments as potential candidate therapeutics against HIV-1. Here we review several bnmAbs and engineered antibody domains (eAds), their in vitro and in vivo antiviral efficacy, mechanisms used by HIV-1 to escape them, and strategies that could be effective to develop more powerful mAb-based HIV-1 therapeutics.

  7. Novel neutralizing monoclonal antibodies protect rodents against lethal filovirus challenges

    Directory of Open Access Journals (Sweden)

    Caleb D. Marceau


    Full Text Available Filoviruses are the causative agents of lethal hemorrhagic fever in human and non-human primates (NHP. The family of Filoviridae is composed of three genera, Ebolavirus, Marburgvirus and Cuevavirus. There are currently no approved vaccines or antiviral therapeutics for the treatment of filovirus infections in humans. Passive transfer of neutralizing antibodies targeting the Ebola virus (EBOV glycoprotein (GP has proven effective in protecting mice, guinea pigs and NHP from lethal challenges with EBOV. In this study, we generated two neutralizing monoclonal antibodies (MAbs, termed S9 and M4 that recognize the GP of EBOV or multiple strains of Marburg virus (MARV, respectively. We characterized the putative binding site of S9 as a linear epitope on the glycan cap of the GP1 subunit of the EBOV-GP. The M4 antibody recognizes an unknown conformational epitope on MARV-GP. Additionally, we demonstrated the post-exposure protection potential of these antibodies in both the mouse and guinea pig models of filovirus infection. These data indicate that MAbs S9 and M4 would be good candidates for inclusion in an antibody cocktail for the treatment of filovirus infections.

  8. Immunohistochemical study of ki-67 and bcl-2 expression in some odontogenic cystic lesions with different clinical behaviors

    Directory of Open Access Journals (Sweden)

    Seyed Hossein Tabatabaei


    Full Text Available Background: Cystic lesions with odontogenic epithelial origin and similar clinicoradiographic appearance, show different clinical behaviors. Objective: To compare some factors related to cell proliferation and escape from apoptosis in epithelium covering two groups of odontogenic cystic lesions with different clinical behaviors. Methods: In this cross-sectional study 11 paraffin-embedded samples were selected of each lesions radicular cyst, dentigerous cyst, odontogenic keratocyst, and unicystic ameloblastoma. The sample underwent immunohistochemical staining for investigating the expression of ki-67 antigen and bcl-2 protein. Data analyzed with SPSS17 software and Kruskal–Wallis and chi-square statistical tests. Findings: Most of ki-67 positive cells were observed in parabasal layer of odontogenic keratocyst [35.50±26.29%; P=0.001]. The average of ki-67-LI was more in parabasal layer of aggressive group (26.80±37.79% compared to non-aggressive group (4.04±3.38%, was not being statistically significant. The highest average of bcl-2-LI was 95±6.70% in basal layer of odontogenic keratocyst (P=0.001. In all layers, the average of bcl-2-LI was more in aggressive lesions compared to non-aggressive ones and the highest amount was found in basal layer (72.45±3.94×10% which was statistically significant (P=0.001. Conclusion: According to the results of this study, more expression of the markers related to escape from apoptosis in aggressive lesions group compared to non-aggressive group, suggests that escape from apoptosis had a more critical role in aggressive behavior of odontogenic cystic lesions.

  9. Moving through three-dimensional phase diagrams of monoclonal antibodies. (United States)

    Rakel, Natalie; Baum, Miriam; Hubbuch, Jürgen


    Protein phase behavior characterization is a multivariate problem due to the high amount of influencing parameters and the diversity of the proteins. Single influences on the protein are not understood and fundamental knowledge remains to be obtained. For this purpose, a systematic screening method was developed to characterize the influence of fluid phase conditions on the phase behavior of proteins in three-dimensional phase diagrams. This approach was applied to three monoclonal antibodies to investigate influences of pH, protein and salt concentrations, with five different salts being tested. Although differences exist between the antibodies, this extensive study confirmed the general applicability of the Hofmeister series over the broad parameter range analyzed. The influence of the different salts on the aggregation (crystallization and precipitation) probability was described qualitatively using this Hofmeister series, with a differentiation between crystallization and precipitation being impossible, however.

  10. Expression of COX-2, iNOS, p53 and Ki-67 in gastric mucosa-associated lymphoid tissue lymphoma

    Institute of Scientific and Technical Information of China (English)

    Hong-Ling Li; Bing-Zhong Sun; Fu-Cheng Ma


    AIM:To assess the expression of cyclooxygenase-2 (COX-2),nitric oxide synthase (iNOS), p53 and Ki-67 in gastric mucosaassociated lymphoid tissue (MALT) lymphoma and clarify the relationship between COX-2 expression and iNOS or p53 expression in these patients.METHODS: The expressions of COX-2, iNOS, p53 and Ki-67 were detected in 32 gastric MALT lymphoma specimens and 10 adjacent mucosal specimens by immunohistochemical Envision method.RESULTS: COX-2 and iNOS expressions were significantly higher in gastric MALT lymphoma tissues than those in adjacent normal tissues. The expression of COX-2 was observed in 22 of 32 cases of MALT lymphoma tissues(68.8%). A positive cytoplasmic immunoreactivity for iNOS was detected in 17 of 31 cases (53.1%). COX-2 expression in gastric MALT lymphoma tissues was positively correlated with iNOS expression (r=0.448, P=0.010) and cell proliferative activity analyzed by Ki-67 labeling index (r=0.410, P=0.020).The expression of COX-2 protein did not correlate with age,sex, stage of disease, lymph node metastasis or differentiation.The accumulation of p53 nuclear phosphoprotein was detected in 19(59.4%) of tumors. p53 protein was expressed in 11 of 23 assessed LG tumors and in 8 of 9 assessed HG tumors.The difference of p53 positivity was found statistically significant between LG and HG cases (P=0.0302). The p53 accumulation correlated with advanced clinical stage (stage Ⅲ+Ⅳ vs stage Ⅰ+Ⅱ, P=-0.017). There was a significant positive correlation between COX-2 expression and p53 accumulation status (r=0.403, P=0.022). The mean PI of Ki-67 in each grade group were 36.0±7.73% in HG and 27.4±9.21% in LG. High-proliferation rate correlated with HG tumors (r=0.419, P=0.017). The correlation coefficient showed a significant positive correlation between PI and COX-2 expression in MALT lymphoma patients (r=0.410,P=0.020).CONCLUSION: COX-2 expresses in the majority of gastric MALT lymphoma tissues and correlates with cellular

  11. Belimumab: anti-BLyS human monoclonal antibody, anti-BLyS monoclonal antibody, BmAb, human monoclonal antibody to B-lymphocyte stimulator. (United States)


    Belimumab is a fully human monoclonal antibody that specifically recognizes and inhibits the biological activity of B-lymphocyte stimulator, or BLyS. Belimumab is in phase III trials for the treatment of systemic lupus erythematosus (SLE) and has completed a phase II trial in rheumatoid arthritis (RA); the product may also have potential in the treatment of other autoimmune disorders. In May 2001, Cambridge Antibody Technology (now MedImmune) completed its discovery programme and Human Genome Sciences identified belimumab as a candidate for clinical development. More than 1000 distinct human antibodies specific to BLyS were characterized by the collaboration.B-lymphocyte stimulator is a naturally occurring protein discovered by Human Genome Sciences that stimulates B-lymphocytes to develop into mature B cells. Laboratory studies have indicated that higher than normal levels of B-lymphocyte stimulator may contribute to the pathogenesis of autoimmune diseases, such as SLE and RA. Human Genome Sciences (HGS) and Cambridge Antibody Technology signed a collaborative agreement in August 1999 to study the B-lymphocyte stimulator as a human protein target. HGS is also developing other BLyS products. In March 2000, HGS and Cambridge Antibody Technology expanded their agreement into a 10-year collaboration and product development alliance, providing Human Genome Sciences with the right to use the antibody technology of Cambridge Antibody Technology to fully develop human antibodies for therapeutic and diagnostic purposes. Cambridge Antibody Technology will receive royalty payments on product sales from HGS, as well as the development and milestone payments it has already received. Belimumab will be manufactured in Human Genome Sciences' manufacturing facility, located in Rockville, MD, USA. HGS holds commercial rights to the drug. In July 2005, GlaxoSmithKline (GSK) exercised its co-development and co-promotion option to belimumab. In an agreement made in June 1996, HGS had

  12. The expression and significance of CSE and Ki-67 in liver tissues in rats with cirrhosis%实验性肝硬化大鼠肝组织中CSE和Ki-67的表达及其意义

    Institute of Scientific and Technical Information of China (English)

    阎继攀; 郑勇; 刘浩; 李睿; 张宁; 齐翠花; 宋丽秀; 陈卫刚


    目的 应用正常及制备的肝硬化大鼠模型的肝脏组织观察H2S过载及不足状态下肝细胞的增殖变化.方法 雌性SD大鼠48只,随机分为6组(每组8只):NF组(正常对照组)、SF组(正常H2S增加组)、PF组(正常H2S减少组)、HF组(肝硬化对照组)、SHF组(肝硬化H2S增加组),PHF组(肝硬化H2S减少组).分组比较门静脉压力及门静脉血浆H2S含量,免疫组化法观察CSE及Ki-67的表达,免疫荧光双重染色观察CSE及Ki-67是否在同一细胞表达.结果 随着给予H2S供体及H2S的抑制剂,HF组H2S含量明显低于NF组(P<0.01),SHF组和SF组中,给予H2S供体均能使H2S含量升高(P<0.01),给予H2S供体使H2S含量进一步升高(P<0.01);HF组CSE表达明显低于NF组(P<0.01),SF组H2S表达较SHF组均减少(P<0.01).H2S对正常大鼠无明显作用;HF组Ki-67表达明显高于NF组(P<0.01),SF组H2S表达较SHF组增加(P<0.01),Ki-67表达在正常大鼠无明显作用.结论 NaHS作为H2S供体可能具有抑制实验性肝硬化大鼠肝组织内细胞增殖的作用,H2S可能通过抑制星状细胞和肝血管平滑肌细胞从而对肝硬化起到保护作用.%Objective To observe the proliferation change of liver cells of experimental cirrhotic rats under excessive H2S environment and inadequate H2S environment. Methods All 48 female SD rats were randomly divided into the following 6 groups ( 8 per group) ; NF group ( normal control group ), SF group ( normal group with increasing H2S), PF group (normal group with decreasing H2S), HF group (cirrhosis group), SHF group (cirrhosis group with increasing H2S), PHF group (cirrhosis group with decreasing H2S). Portal venous pressure and the hydrogen sulfide density in portal vein plasma among groups were compared; CSE and Ki-67 expression were observed by immunohisto-chemistry; CSE and Ki-67 expression in the same cell were observed by double immunofluorescence staining. Results With the supply of H2S donor and H2S

  13. The role of Ki-67 in women with a resistant prolactinoma: a retrospective analysis in 199 hospitalized patients over a period of 5 years. (United States)

    Lu, Chao; Ren, Zongming; Huan, Cheng; Cui, Guihua


    Proliferation-associated antigen Ki-67 is used for the histological evaluation of different tumors. Few studies have been conducted on women with a resistant prolactinoma. To better define the characteristics and to evaluate the differences between patients with different Ki-67 labeling index (LI), a retrospective study was designed to recruit 199 females with a resistant prolactinoma. The patients were divided into two groups, patients with Ki-67 LI≥3% and patients with Ki-67 LI3% group were also larger (p=0.043), had a higher rate of invasion (p=0.014), and were associated with more frequent polyuria and polydipsia (p=0.008) compared to the LI3% group remained significantly higher compared to patients with LI3% group were also significantly higher (p=0.037, p=0.041). Additionally, the postoperative PRL normalization rate was lower in patients with LI>3% compared with patients with LI3% and LIprolactinoma.

  14. Monoclonal antibodies directed to fucoidan preparations from brown algae. (United States)

    Torode, Thomas A; Marcus, Susan E; Jam, Murielle; Tonon, Thierry; Blackburn, Richard S; Hervé, Cécile; Knox, J Paul


    Cell walls of the brown algae contain a diverse range of polysaccharides with useful bioactivities. The precise structures of the sulfated fucan/fucoidan group of polysaccharides and their roles in generating cell wall architectures and cell properties are not known in detail. Four rat monoclonal antibodies, BAM1 to BAM4, directed to sulfated fucan preparations, have been generated and used to dissect the heterogeneity of brown algal cell wall polysaccharides. BAM1 and BAM4, respectively, bind to a non-sulfated epitope and a sulfated epitope present in the sulfated fucan preparations. BAM2 and BAM3 identified additional distinct epitopes present in the fucoidan preparations. All four epitopes, not yet fully characterised, occur widely within the major brown algal taxonomic groups and show divergent distribution patterns in tissues. The analysis of cell wall extractions and fluorescence imaging reveal differences in the occurrence of the BAM1 to BAM4 epitopes in various tissues of Fucus vesiculosus. In Ectocarpus subulatus, a species closely related to the brown algal model Ectocarpus siliculosus, the BAM4 sulfated epitope was modulated in relation to salinity levels. This new set of monoclonal antibodies will be useful for the dissection of the highly complex and yet poorly resolved sulfated polysaccharides in the brown algae in relation to their ecological and economic significance.

  15. Monoclonal antibodies directed to fucoidan preparations from brown algae.

    Directory of Open Access Journals (Sweden)

    Thomas A Torode

    Full Text Available Cell walls of the brown algae contain a diverse range of polysaccharides with useful bioactivities. The precise structures of the sulfated fucan/fucoidan group of polysaccharides and their roles in generating cell wall architectures and cell properties are not known in detail. Four rat monoclonal antibodies, BAM1 to BAM4, directed to sulfated fucan preparations, have been generated and used to dissect the heterogeneity of brown algal cell wall polysaccharides. BAM1 and BAM4, respectively, bind to a non-sulfated epitope and a sulfated epitope present in the sulfated fucan preparations. BAM2 and BAM3 identified additional distinct epitopes present in the fucoidan preparations. All four epitopes, not yet fully characterised, occur widely within the major brown algal taxonomic groups and show divergent distribution patterns in tissues. The analysis of cell wall extractions and fluorescence imaging reveal differences in the occurrence of the BAM1 to BAM4 epitopes in various tissues of Fucus vesiculosus. In Ectocarpus subulatus, a species closely related to the brown algal model Ectocarpus siliculosus, the BAM4 sulfated epitope was modulated in relation to salinity levels. This new set of monoclonal antibodies will be useful for the dissection of the highly complex and yet poorly resolved sulfated polysaccharides in the brown algae in relation to their ecological and economic significance.

  16. Fingerprinting of Natural Product by Eastern Blotting Using Monoclonal Antibodies

    Directory of Open Access Journals (Sweden)

    Hiroyuki Tanaka


    Full Text Available We succeeded in developing the fingerprint of natural product by eastern blotting using monoclonal antibodies. After developing and separating them on a TLC plate, solasodine glycosides are oxidized by NaIO4 and reacted with a protein to give conjugates which are recognized with anti-solamargine monoclonal antibody (MAb. Anti-solamargine MAb having wide cross-reactivity can stain and detect all solasodine glycosides by fingerprint. Different sensitivity between solamargine and solasonine was observed. The detection limit was 1.6 ng of solasonine. The hydrolysed products of solamargine were determined by fingerprint of eastern blotting compared to their Rf values depending on the sugar number. Fingerprint by eastern blotting using anti-ginsenoside Rb1 MAb distinguished the formula containing ginseng prescribed in traditional Chinese medicine. By double-staining of ginsenosides it is possible to suggest that the staining color shows the pharmacological activity, such as the purple bands indicate ginsenosides having stimulation activity, and the blue color indicated compound like ginsenosides possessed the depression affect for the central nervous system (CNS, respectively.

  17. [Obtaining monoclonal antibodies against outer membrane glycoproteins of Entamoeba histolytica]. (United States)

    Agundis, C; Isibasi, A; Ortíz, V; Reyes, J L; Paniagua, J; Ramírez, A; Kumate, J


    The goal of this paper was the production of monoclonal antibodies capable of detecting relevant antigens from the surface of Entamoeba histolytica trophozoites, with the purpose of using them as a diagnostic test. The cellular fusion for obtaining the monoclonal antibodies (mAb) was done with spleen cells from BALB/c mice, previously immunized with glycoproteins from the membrane, as well as Sp2/0 cells. The hybridoma supernatants were tested with ELISA, using glycoproteins and lipopeptide phosphoglycans (LPPG) as antigens. Seven hybridomas producing mAb against the glycoproteins were found. Among these, three recognize LPPG. The ability of reacting with the mAb against two molecules disappeared for all the LPPG positive ones when were treated with meta-periodate, and only three reacted against the glycoproteins. All of the mAb were of the Ig M isotypes. They were characterized by Dot blot and Western blot assays. From the results, one may deduce that some mAb recognize as epitopes the polysaccharide portion, and thus infer that they are directed of against the surface and therefore, in the future, could be used with a diagnostic purpose.

  18. Immunotherapy of hepatoma with a monoclonal antibody against murine endoglin

    Institute of Scientific and Technical Information of China (English)

    Guang-Hong Tan; Feng-Ying Huang; Hua Wang; Yong-Hao Huang; Ying-Ying Lin; Yue-Nan Li


    AIM: To explore the capability of a monoclonal antibody(mAb) against murine endoglin to inhibit tumor angiogenesis and suppression of hepatoma growth in murine models.METHODS: A monoclonal antibody against murine endoglin was purified by affinity chromatography and passively transfused through tail veins in two murine hepatoma models. Tumor volume and survival time were observed at three-day intervals for 48 d. Microvessels in tumor tissues were detected by immunohistochemistry against CD31, and angiogenesis in vivo was determined by alginate encapsulated assay. In addition, tumor cell apoptosis was detected by TUNEL assay.RESULTS: Passive immunotherapy with anti-endoglin mAb could effectively suppress tumor growth, and prolonged the survival time of hepatoma-bearing mice.Angiogenesis was apparently inhibited within the tumor tissues, and the vascularization of alginate beads was also reduced in the mice passively transfused with antiendoglin mAb. In addition, increased apoptotic cells were observed within the tumor tissues from the mice passively transfused with anti-endoglin mAb.CONCLUSION: Passive immunotherapy with antiendoglin mAb effectively inhibits tumor growth via inhibiting tumor angiogenesis and increasing tumor cell apoptosis, which may be highly correlated with the blockage of endoglin-related signal pathway induced by anti-endoglin mAb.

  19. Application of the Expression of P16 and Ki67 in Diagnosis of Cervical Le-sions%P16与Ki67的表达在子宫颈病变诊断中的应用

    Institute of Scientific and Technical Information of China (English)



    目的 探究P16与Ki67的表达在子宫颈病变诊断中的应用.方法 采用回顾性分析的方法,选取2014年8月—2015年12月期间在该院妇科进行治疗的90例子宫颈病变患者作为该研究的对象,其中30例为低级别鳞状上皮内病变(LSIL)(包括CINI,扁平湿疣,尖锐湿疣),30例为高级别鳞状上皮内病变(HSIL)(CINII10例,CINIII20例)、30例子宫颈良性反应性病变(包括不成熟鳞化,上皮萎缩,修复性上皮增生)患者,采用免疫组化SP法对这90例患者的P16以及Ki67表达情况进行检测分析.结果经过HE染色分析,可以发现Ki67表达阳性细胞在宫颈鳞状上皮中的位置分布与子宫颈上皮内瘤变有着一定的相关性,LSIL多分布于下1/3的位置,CINII多分布于下1/3~2/3的位置,而CINIII多分布于>2/3位置,子宫颈良性反应性病变分布于下1/3的基底和副基底层;P16的表达水平在LSIL、HSIL、子宫颈良性反应性病变中的差异有统计学意义(P=0.021<0.05).且P16的表达水平与子宫颈上皮内瘤变的级别呈现出正相关的关系,差异有统计学意义(P=0.013<0.05).结论 P16与Ki67的表达可水平可用于子宫颈良性反应性病变的甄别,还与子宫颈上皮内瘤变的进展联系密切,对P16以及Ki67表达水平的联合检测能够帮助子宫颈病变诊断,可以在临床医学中得以广泛的推广应用.

  20. Reproducibility of the Nottingham modification of the Scarff-Bloom-Richardson histological grading system and the complementary value of Ki-67 to this system

    Institute of Scientific and Technical Information of China (English)

    ZHANG Rui; CHEN Hui-jiao; WEI Bing; ZHANG Hong-ying; PANG Zong-guo; ZHU Hong; ZHANG Zhang; FU Jing; BU Hong


    Background The reproducibility of the Nottingham modification of the Scarff-Bloom-Richardson (NSBR) histological grading system for invasive breast cancer (IBC) adopted by the World Health Organization (WHO) has previously not been studied in Chinese hospitals. The proliferation marker, Ki-67, has been widely applied in detecting IBC. The objective of this study was to assess the reproducibility of the NSBR system among Chinese pathologists and the complementary value that Ki-67 brings to this system.Methods Four general pathologists graded 100 IBC cases independently, which had previously been graded by specialists in breast pathology. The interobserver reproducibility among four general pathologists and pairwise reproducibility between each of them and the specialists were assessed. The Ki-67 labeling index (Ki-67LI) was determined by immunohistochemistry, and its correlations with histological grade and survival were determined.Results With respect to interobserver reproducibility, NSBR grading was fairly reproducible (kappa=0.34); as for the components of NSBR grading, agreement was best for tubule formation (kappa=0.46), intermediate for nuclear pleomorphism (kappa=0.42), and poorest for mitotic count (kappa=0.28). In terms of pairwise reproducibility, agreement was fair to substantial with NSBR grading (kappa=0.30-0.69) and nuclear pleomorphism (kappa=0.28-0.69), moderate to substantial for tubule formation (kappa=0.51-0.78), and slight to substantial for mitotic count (kappa=0.19-0.71).There were characteristic Ki-67LI ranges for grades 1,2 and 3 tumors. Univariate analysis showed that Ki-67 was able to divide grade 2 patients into two different prognostic subgroups. Multivariate analysis of grade 2 patients with negative lymph node demonstrated that Ki-67 was an independent prognosticator for overall survival.Conclusions The reproducibility of grading by general pathologists could be enhanced. Specialization in breast pathology is essential for accurate

  1. 胃腺癌组织中 p53表达与分化程度、Ki-67表达的关系

    Institute of Scientific and Technical Information of China (English)

    贾振军; 李其海


    目的:探讨胃腺癌组织中p53表达与分化程度、Ki-67表达的相关性。方法采用免疫组化SP法检测130例胃癌组织中p53、Ki-67的表达。分析胃腺癌组织中p53表达与胃癌分化程度、Ki-67的表达的相关性。结果本组标本中p53阳性72例(55.38%), Ki-67阳性58例(44.62%)。本组胃腺癌为低分化者p53阳性63例(72.41%),中高分化者p53阳性9例(20.93%),二者p53阳性表达率相比P<0.05。本组标本p53阳性者中Ki-67阳性69例(95.83%),p53阴性者中Ki-67阳性40例(68.97%),p53阳性、阴性者Ki-67阳性率相比P<0.05。结论胃腺癌组织中p53阳性表达与分化程度和Ki-67表达有关。

  2. NCI Requests Targets for Monoclonal Antibody Production and Characterization - Office of Cancer Clinical Proteomics Research (United States)

    In an effort to provide well-characterized monoclonal antibodies to the scientific community, NCI's Antibody Characterization Program requests cancer-related protein targets for affinity production and distribution.

  3. [Production and characteristics of monoclonal antibodies to the diphtheria toxin]. (United States)

    Valiakina, T I; Lakhtina, O E; Komaleva, R L; Simonova, M A; Samokhvalova, L V; Shoshina, N S; Kalinina, N A; Rubina, A Iu; Filippova, M A; Vertiev, Iu V; Grishin, E V


    Monoclonal antibodies to the diphtheria toxin were produced without cross reactivity with the thermolabile toxin (LT) from Escherichia coli; ricin; choleraic toxin; the SeA, SeB, SeE, SeI, and SeG toxins of staphylococcus; the lethal factor of the anthrax toxin; and the protective antigen of the anthrax toxin. A pair of antibodies for the quantitative determination of the diphtheria toxin in the sandwich variation of enzyme-linked immunosorbent assay (ELISA) was chosen. The determination limit of the toxin was 0.7 ng/ml in plate and 1.6 ng/ml in microchip ELISA. The presence of a secretion from the nasopharynx lavage did not decrease the sensitivity of the toxin determination by sandwich ELISA. The immunization of mice with the diphtheria toxin and with a conjugate of the diphtheria toxin with polystyrene microspheres demonstrated that the conjugate immunization resulted in the formation of hybridoma clones which produced antibodies only to the epitopes of the A fragment of the diphtheria toxin. The immunization with the native toxin caused the production of hybridoma clones which predominantly produced antibodies to the epitopes of the B fragment.

  4. [Monoclonal antibodies against PCSK9: from bench to clinic]. (United States)

    Guijarro Herraiz, Carlos


    Antibodies are glycoproteins with high specificity binding to multiple antigens due to the large number of structural conformations of the variable chains. Hybridoma technology (fusion of myeloma cells with immunoglobulin-producing lymphocytes) has allowed the synthesis of large quantities of unique antibodies (monoclonal [mAb]). mAbs were initially murine. Subsequently, chimeric mAbs were developed, followed by humanized mAbs and finally human mAbs. The high selectivity and good tolerance of human mAbs allows their therapeutic administration to block specific exogenous or endogenous molecules. Selective human mAbs to the catalytic domain of PCSK9 have recently been developed. These antibodies block PCSK9, favour low-density lipoprotein receptor recycling and markedly reduce circulating cholesterol. Preliminary studies indicate that lowering cholesterol through anti-PCSK9 antibodies may significantly reduce the cardiovascular complications of arteriosclerosis. Copyright © 2016 Elsevier España, S.L.U. y Sociedad Española de Arteriosclerosis. All rights reserved.

  5. Expressions of survivin, P16(INK4a), COX-2, and Ki-67 in cervical cancer progression reveal the potential clinical application. (United States)

    Zhou, W Q; Sheng, Q Y; Sheng, Y H; Hou, W J; Xu, G X; Wu, Y M; Lu, H


    To explore the significance of survivin, P16(INK4a), COX-2, and Ki-67 expressions for prediction of cervical cancer progression. A retrospective study was performed in 129 cases including 24 squamous carcinoma of the cervix (SCC), 70 cervical intraepithelial neoplasias (CIN), 15 cervical condyloma acuminatum (CCA), ten chronic cervicitis (CC), and ten normal cervix (NC). Protein expressions were evaluated using immunohistochemistry. Survivin, P16(INK4a); COX-2, and Ki-67 were highly expressed in SCC and CIN compared with others. Their expression rates were gradually increased in CIN I, CIN II, CIN III, and SCC groups, showing 72.00%, 88.00%, 90.00%, and 95.83% for P16(INK4a), 68.00%, 84.00%, 95.00% and 100.00% for COX-2, 76.00%, 96.00%, 100.00%, and 100.00 for Ki-67, respectively. There were significant correlations between survivin and P16(INK4a), COX-2, Ki-67, as well as P16(INK4a) and Ki-67. Survivin, P16(INK4a), COX-2 and Ki-67 play critical roles for development and progression of cervical cancer.

  6. An Advanced Deep Learning Approach for Ki-67 Stained Hotspot Detection and Proliferation Rate Scoring for Prognostic Evaluation of Breast Cancer. (United States)

    Saha, Monjoy; Chakraborty, Chandan; Arun, Indu; Ahmed, Rosina; Chatterjee, Sanjoy


    Being a non-histone protein, Ki-67 is one of the essential biomarkers for the immunohistochemical assessment of proliferation rate in breast cancer screening and grading. The Ki-67 signature is always sensitive to radiotherapy and chemotherapy. Due to random morphological, color and intensity variations of cell nuclei (immunopositive and immunonegative), manual/subjective assessment of Ki-67 scoring is error-prone and time-consuming. Hence, several machine learning approaches have been reported; nevertheless, none of them had worked on deep learning based hotspots detection and proliferation scoring. In this article, we suggest an advanced deep learning model for computerized recognition of candidate hotspots and subsequent proliferation rate scoring by quantifying Ki-67 appearance in breast cancer immunohistochemical images. Unlike existing Ki-67 scoring techniques, our methodology uses Gamma mixture model (GMM) with Expectation-Maximization for seed point detection and patch selection and deep learning, comprises with decision layer, for hotspots detection and proliferation scoring. Experimental results provide 93% precision, 0.88% recall and 0.91% F-score value. The model performance has also been compared with the pathologists' manual annotations and recently published articles. In future, the proposed deep learning framework will be highly reliable and beneficial to the junior and senior pathologists for fast and efficient Ki-67 scoring.

  7. Prognostic value of tissue protein expression levels of MIB-1 (Ki-67) in Danish ovarian cancer patients. From the 'MALOVA' ovarian cancer study

    DEFF Research Database (Denmark)

    Heeran, Mel C; Høgdall, Claus K; Kjaer, Susanne K


    The primary objective of this study was to assess the expression of MIB-1 (Ki-67) in tumour tissues from 808 patients with epithelial ovarian tumours. The second was to evaluate, whether MIB-1 (Ki-67) tissue expression levels correlate with clinicopathological parameters and prognosis...... of the disease. Using tissue arrays (TA), we analysed the MIB-1 (Ki-67) expression levels in tissues from 202 women with borderline ovarian tumours (BOT) (177 stage I, 5 stage II, 19 stage III, 1 stage IV) and 606 ovarian cancer (OC) patients (177 stage I, 64 stage II, 311 stage III, 54 stage IV). Using a 10......% cut-off level for MIB-1 (Ki-67) overexpression, 12% of the BOTs and 51% of the OCs were positive for MIB-1 (Ki-67) expression. The frequency of MIB-1 (Ki-67) expression-positive OC increased with increasing FIGO stage (p = 0.003), increasing histological grade (p ≤ 0.0001), and a significantly...

  8. Antibodies in infectious diseases: polyclonals, monoclonals and niche biotechnology. (United States)

    Berry, Jody D; Gaudet, Ryan G


    Antibody preparations have a long history of providing protection from infectious diseases. Although antibodies remain the only natural host-derived defense mechanism capable of completely preventing infection, as products, they compete against inexpensive therapeutics such as antibiotics, small molecule inhibitors and active vaccines. The continued discovery in the monoclonal antibody (mAb) field of leads with broadened cross neutralization of viruses and demonstrable synergy of antibody with antibiotics for bacterial diseases, clearly show that innovation remains. The commercial success of mAbs in chronic disease has not been paralleled in infectious diseases for several reasons. Infectious disease immunotherapeutics are limited in scope as endemic diseases necessitate active vaccine development. Also, the complexity of these small markets draws the interest of niche companies rather than big pharmaceutical corporations. Lastly, the cost of goods for mAb therapeutics is inherently high for infectious agents due to the need for antibody cocktails, which better mimic polyclonal immunoglobulin preparations and prevent antigenic escape. In cases where vaccine or convalescent populations are available, current polyclonal hyperimmune immunoglobulin preparations (pIgG), with modern and highly efficient purification technology and standardized assays for potency, can make economic sense. Recent innovations to broaden the potency of mAb therapies, while reducing cost of production, are discussed herein. On the basis of centuries of effective use of Ab treatments, and with growing immunocompromised populations, the question is not whether antibodies have a bright future for infectious agents, but rather what formats are cost effective and generate safe and efficacious treatments to satisfy regulatory approval. Copyright © 2011 Elsevier B.V. All rights reserved.

  9. P16(INK 4a) and Ki-67 expression in human papilloma virus-related head and neck mucosal lesions. (United States)

    Gültekin, Sibel Elif; Sengüven, Burcu; Klussmann, Jens Peter; Dienes, Hans Peter


    Human papilloma virus (HPV) is postulated as a risk factor in the etiology of some specific mucosal pathologies in the head and neck regions. Despite the frequent use of p16(INK4a) as a surrogate marker for HPV-infection, there is still controversy with respect to its reliability. This study has been undertaken to assess the potential role of p16(INK 4a) and Ki-67 expression in HPV-related lesions. The study was conducted on 71 specimens of oral, tonsillar and laryngeal lesions which comprised 25 dysplasia and 46 papilloma specimens. Specimens were immunohistochemically stained for p16(INK4A) and Ki-67 proteins. HPV DNA was determined by one step multiplex polymerase chain reaction. HPV DNA was detected in 33.8% of all lesions. Tonsil and larynx lesions showed significant differences with oral lesions for HPV positivity (p < 0.001). p16(INK 4a) over-expression was seen in 56.5% of papilloma and 60% of dysplasia specimens. HPV status showed a positive correlation with p16(INK 4a) expression in tonsillar dysplasias (p < 0.001). p16(INK 4a) expression may have a value as a marker in high risk HPV induced dysplasias, but not in low risk infected lesions. The proliferation index is not related to HPV-induced lesions and may be evaluated as an independent marker in head and neck premalignant lesions.

  10. Metachronous multicentric giant-cell tumor of the bone in the lower limb. Case report and Ki-67 immunohistochemistry study. (United States)

    Rousseau, Marc-Antoine; Handra-Luca, Adriana; Lazennec, Jean-Yves; Catonné, Yves; Saillant, Gérard


    Multicentric giant-cell tumors of the bone (GCTs) are rare. Little is known about the mechanisms by which these tumors spread and how 1% of GCT turn out to be multicentric. We report the case of a 19-year-old woman with metachronous multiple and recurrent GCTs that were unusual in their pattern of progression along the right lower limb over a 23-year period. Histology showed no evidence of malignant transformation. The treatment was repeated curettage and packing with cement. This did not permit a wide surgical margin, but avoided amputation and preserved full limb function. We tested the proliferation index marker Ki-67 in the tumor specimens. Ki-67 expression was limited to the mononuclear cell component of the tumors. The proliferation index was similar in each new tumor and higher in recurrences for each location. In this case, proliferation was initially low in the new tumor location, despite the time difference and independent from the initial clone evolution. Proliferation index increased in recurrent GCTs after marginal margin resection.

  11. Immunoexpression of Ki67, proliferative cell nuclear antigen, and Bcl-2 proteins in a case of ameloblastic fibrosarcoma. (United States)

    Pontes, Hélder Antônio Rebelo; Pontes, Flávia Sirotheau Corrêa; Silva, Brunno Santos de Freitas; Cury, Sérgio Elias Vieira; Fonseca, Felipe Paiva; Salim, Rodrigo Alves; Pinto Júnior, Décio dos Santos


    Ameloblastic fibrosarcoma (AFS), regarded as the malignant counterpart of the benign ameloblastic fibroma, is an extremely rare odontogenic neoplasm with only 68 cases reported in the English literature up to 2009. It is composed of a benign odontogenic epithelium, resembling that of ameloblastoma, and a malignant mesenchymal part exhibiting features of fibrosarcoma. Due to the rarity of the lesion, little is known about its molecular pathogenesis; therefore, in the current study, we sought to evaluate the immunoexpression of Ki67, proliferative cell nuclear antigen, and Bcl-2 proteins in AFS, comparing the results obtained with its benign counterpart, as well as to report a new case of this rare entity affecting a 19-year-old female patient. The results obtained revealed that all the proteins evaluated were overexpressed in the malignant mesenchymal portion of AFS if compared with ameloblastic fibroma, suggesting that nuclear proliferative factors such as Ki67 and proliferative cell nuclear antigen, in association to histopathologic features, may be useful markers for identifying the malignancy and that, despite the lack of molecular analysis in the case reported, Bcl-2 alteration may play a role in AFS pathogenesis. Copyright © 2010 Elsevier Inc. All rights reserved.

  12. Multiple Bone Metastasis of Sclerosing Epithelioid Fibrosarcoma 12 Years after Initial Surgery—Increasing Ki-67 Labeling Index

    Directory of Open Access Journals (Sweden)

    Atsuko Kanno


    Full Text Available Sclerosing epithelioid fibrosarcoma (SEF is a rare sarcoma of low-grade malignancy. There has been no report to describe the comparison of histological features of SEF between primary and metastatic lesions in spite of high local recurrence rate. We report the histological changes and increasing Ki-67 labeling index of the primary and metastatic lesions of SEF. The patient was a 31-year-old man. At 18, a tumor in the abdominal wall was excised. At 23, the tumor recurred which was removed again. At 30, he was referred to our hospital because of swelling and pain in the chest. Histological examination of the chest wall tumor showed epithelioid cells arranged like alveolar pattern with dense collagen stroma. These findings were consistent with those of SEF. Abdominal and the rib tumors showed the same immunohistochemistrical expression. It is noteworthy that the tumor cells of the rib lesion showed increased cellularity, and its Ki-67 activity was higher as compared with the abdominal tumor, suggestive of progression of malignancy of SEF.

  13. Characterization and evaluation of monoclonal antibodies developed for typing influenza A and influenza B viruses.


    Walls, H H; Harmon, M W; Slagle, J J; Stocksdale, C; Kendal, A P


    Monoclonal antibodies that are broadly reactive with influenza A or influenza B viruses were produced as stable reagents for typing influenza viruses. Monoclonal antibodies to influenza A were specific for either matrix protein or nucleoprotein. The antibodies to influenza B were specific for nucleoprotein or hemagglutinin protein. In an enzyme immunoassay procedure, influenza A antibodies detected H1N1, H2N2, and H3N2 influenza A virus strains collected between 1934 and 1984. Each of the inf...

  14. New monoclonal antibodies directed against human renin. Powerful tools for the investigation of the renin system.


    Galen, F X; Devaux, C.; Atlas, S; Guyenne, T; Menard, J; Corvol, P; Simon, D.; Cazaubon, C; Richer, P; Badouaille, G


    Monoclonal antibodies directed against human renin were obtained by the fusing of myeloma cells with spleen cells from Balb/c or high-responder Biozzi mice injected with pure tumoral or highly purified renal renin. These procedures resulted in the production of seven stable monoclonal antibodies to human renin. Antibodies in the hybridoma culture medium were screened by binding to pure iodinated renin or insolubilized renin in a solid phase assay. The concentration of purified antibodies that...

  15. Preparation of monoclonal antibody to P53 and its clinical application

    Institute of Scientific and Technical Information of China (English)

    Wenqing Wei; Junhua Wu; Jing Liu; Yuxia Wang


    Objective:The aim of this study was to prepare monoclonal antibody against P53, a kind of tumor suppressor protein,and use the antibody initial y in clinical immunoassay. Methods:Monoclonal antibody was prepared and identified via the classic protocol of monoclonal antibody preparation. Identified monoclonal antibodies were purified by af inity chro-matography. Antibody titer was determined by enzyme linked immunosorbent assay (ELISA). The specific binding activity of antibody was detected by Western blotting and immunohistochemistry. Results:Three strains of monoclonal antibodies named 1P15, 2P37 and 3P40 were obtained and purified by af inity chromatography. The purity of antibodies was higher than 90%. The titers of antibodies were more than 1:6000. Western blot and immunohistochemistry assay showed that the specific antibody can combine with endogenous P53 protein in the tumor celllines and determine the expression of P53 in tumor tis-sue. Conclusion:Three strains of monoclonal antibodies with high af inity to P53 were successful y established, which can be used for detecting the expression of P53 in tumor cells or tissue.

  16. Monoclonal antibodies against a synthetic peptide from human immunodeficiency virus type 1 Nef protein

    DEFF Research Database (Denmark)

    Steinaa, L; Wulff, A M; Saermark, T


    Monoclonal antibodies against a synthetic peptide (aa 138-152) from HIV-1 Nef protein were produced and characterized. Three hybridoma lines producing monoclonal antibodies (MAbs) against the synthetic peptide were generated by fusion between P3-X63 Ag8.653 myeloma cells and BALB/c splenocytes fr...

  17. Preparation of Europium Induced Conformation—specific anti—calmodulin Monoclonal Antibody

    Institute of Scientific and Technical Information of China (English)

    WeiGuoLI; ChaoQI; 等


    Monoclonal antibody technique was employed to detect the conformational difference of CaM induced by metal ions. A trivalent europium ion induced conformation-specific anti-calmodulin monoclonal antibody was successfully prepared with europium-saturated calmodulin as antigen.

  18. Preparation of Europium Induced Conformation-specific anti-calmodulin Monoclonal Antibody

    Institute of Scientific and Technical Information of China (English)


    Monoclonal antibody technique was employed to detect the conformational difference of CaM induced by metal ions. A trivalent europium ion induced conformation-specific anti-calmodulin monoclonal antibody was successfully prepared with europium-saturated calmodulin as antigen.

  19. Development and characterization of mouse monoclonal antibodies specific for chicken interleukin 18 (United States)

    Four mouse monoclonal antibodies (mAbs) which are specific for chicken interleukin 18 (chIL18) were produced and characterized by enzyme-linked immunosorbent assay (ELISA), Western blotting, quantitative real-time PCR and neutralization assays. Monoclonal antibodies specific for chIL18 identified a ...

  20. Reactivity of eleven anti-human leucocyte monoclonal antibodies with lymphocytes from several domestic animals

    DEFF Research Database (Denmark)

    Aasted, Bent; Blixenkrone-Møller, Merete; Larsen, Else Bang


    Nine commercially available monoclonal antibodies and two monoclonal antibodies from The American Type Culture Collection, raised against various human leucocyte surface antigens, were tested on lymphocytes from cow, sheep, goat, swine, horse, cat, dog, mink, and rabbit as well as man. Four...... antibodies bound to lymphocytes from some of the animals. These were the antibodies against CD8 and CD4 antigen, the antibody to C3b-receptor, and the antibody to the HLA-DR antigen. The CD8 antigen-reactive antibody reacted with lymphocytes from mink, cat, dog, and sheep, while the CD4 antigen...

  1. Discovery of functional monoclonal antibodies targeting G-protein-coupled receptors and ion channels. (United States)

    Wilkinson, Trevor C I


    The development of recombinant antibody therapeutics is a significant area of growth in the pharmaceutical industry with almost 50 approved monoclonal antibodies on the market in the US and Europe. Despite this growth, however, certain classes of important molecular targets have remained intractable to therapeutic antibodies due to complexity of the target molecules. These complex target molecules include G-protein-coupled receptors and ion channels which represent a large potential target class for therapeutic intervention with monoclonal antibodies. Although these targets have typically been addressed by small molecule approaches, the exquisite specificity of antibodies provides a significant opportunity to provide selective modulation of these target proteins. Given this opportunity, substantial effort has been applied to address the technical challenges of targeting these complex membrane proteins with monoclonal antibodies. In this review recent progress made in the strategies for discovery of functional monoclonal antibodies for these challenging membrane protein targets is addressed.

  2. Is Ki-67 Expression Prognostic for Local Relapse in Early-Stage Breast Cancer Patients Treated With Breast Conservation Therapy (BCT)?

    Energy Technology Data Exchange (ETDEWEB)

    Hafeez, Farhaan [Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, Connecticut (United States); Neboori, Hanmanth J. [Drexel Medical College, Philadelphia, Pennsylvania (United States); Harigopal, Malini [Department of Pathology, Yale University School of Medicine, New Haven, Connecticut (United States); Wu, Hao; Haffty, Bruce G. [Department of Radiation Oncology, University of Medicine and Dentistry of New Jersey–Robert Wood Johnson School of Medicine, New Brunswick, New Jersey (United States); Yang, Qifeng [Department of Breast Surgery, Shandong University School of Medicine, Shanghai (China); Schiff, Devora [Department of Radiation Oncology, University of Medicine and Dentistry of New Jersey–Robert Wood Johnson School of Medicine, New Brunswick, New Jersey (United States); Moran, Meena S., E-mail: [Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, Connecticut (United States)


    Purpose: Ki-67 is a human nuclear protein whose expression is strongly up-regulated in proliferating cells and can be used to determine the growth fraction in clonal cell populations. Although there are some data to suggest that Ki-67 overexpression may be prognostic for endpoints such as survival or postmastectomy recurrence, further elucidation of its prognostic significance is warranted. Specifically after breast conservation therapy (BCT) (defined in this setting as breast-conserving surgery and adjuvant radiation therapy), whether Ki-67 predicts for locoregional recurrence has not been investigated. The purpose of this study was to assess Ki-67 expression in a cohort of early-stage breast cancer patients to determine whether a significant independent association between Ki-67 and locoregional relapse exists. Methods and Materials: Ki-67 staining was conducted on a tissue microarray of 438 patients previously treated with BCT, and expression was analyzed with clinicopathologic features and outcomes from our database. Results: Ki-67 expression was more prevalent in black patients (37% of black patients vs 17% of white patients, P<.01), younger patients (27% of patients aged ≤50 years vs 15% of patients aged >50 years, P<.01), estrogen receptor (ER)–negative tumors (25% of ER-negative tumors vs 17% of ER-positive tumors, P=.04), human epidermal growth factor receptor 2 (HER2)/neu–positive tumors (35% of HER2-positive tumors vs 18% of HER2-negative tumors, P=.01), and larger tumors (26% of T2 tumors vs 16% of T1 tumors, P=.03). On univariate/multivariate analysis, Ki-67 did not predict for overall survival (74.4% vs 72.6%), cause-specific survival (82.9% vs 82.1%), local relapse-free survival (83.6% vs 88.5%), distant metastasis-free survival (76.1% vs 81.4%), recurrence-free survival (65.5% vs 74.6%), and locoregional recurrence-free survival (81.6% vs 84.7%): P>.05 for all. Conclusions: Ki-67 appears to be a surrogate marker for aggressive disease and

  3. Correlation analysis between expression of PCNA, Ki-67 and COX-2 and X-ray features in mammography in breast cancer. (United States)

    Qiu, Xiaoming; Mei, Jixin; Yin, Jianjun; Wang, Hong; Wang, Jinqi; Xie, Ming


    This study investigated expression of proliferating cell nuclear antigen (PCNA), proliferation-associated nuclear antigen (Ki-67) and cyclooxygenase-2 (COX-2) in tissues of breast invasive ductal carcinoma, and analyzed the correlations between these indexes and X-ray features in mammography. A total of 90 patients who were admitted to Huangshi Central Hospital and diagnosed as breast invasive ductal carcinoma from January 2014 to January 2016 were selected. The expression of PCNA, Ki-67 and COX-2 in cancer tissues and cancer-adjacent normal tissues of patients were detected by immunohistochemical staining, and X-ray features in mammography of patients were observed. By using Spearman correlation analysis, the correlations between expression of PCNA, Ki-67 and COX-2 and X-ray features in mammography in breast cancer were investigated. As a result, the positive expression rates of PCNA, Ki-67 and COX-2 in cancer tissues of the patient groups were respectively 42.2, 45.6 and 51.1%, which were significantly higher than those in cancer-adjacent normal tissues of the control group (pcorrelation with age and tumor size (p>0.05). PCNA, Ki-67 and COX-2 expression in cancer tissues of the patient group had no correlation with the existence of lumps and localized density-increased shadows (p>0.05), but were associated with manifestations of architectural distortion, calcification as well as skin and nipple depression (pcorrelation analysis revealed that there was a significantly positive correlation between the expression of PCNA and COX-2 in cancer tissues of the patient group (r=0.676, pcorrelation between the expression of Ki-67 and COX-2 (r=0.724, pcorrelation with the expression of Ki-67 (p>0.05). In conclusion, PCNA, Ki-67 and COX-2 expression is of great significance in the occurrence, invasion and metastasis of breast invasive ductal carcinoma. There is a strong correlation between PCNA, Ki-67 and COX-2 expression levels and X-ray features in mammography in breast

  4. Distinguishing Low-Risk Luminal A Breast Cancer Subtypes with Ki-67 and p53 Is More Predictive of Long-Term Survival. (United States)

    Lee, Se Kyung; Bae, Soo Youn; Lee, Jun Ho; Lee, Hyun-Chul; Yi, Hawoo; Kil, Won Ho; Lee, Jeong Eon; Kim, Seok Won; Nam, Seok Jin


    Overexpression of p53 is the most frequent genetic alteration in breast cancer. Recently, many studies have shown that the expression of mutant p53 differs for each subtype of breast cancer and is associated with different prognoses. In this study, we aimed to determine the suitable cut-off value to predict the clinical outcome of p53 overexpression and its usefulness as a prognostic factor in each subtype of breast cancer, especially in luminal A breast cancer. Approval was granted by the Institutional Review Board of Samsung Medical Center. We analyzed a total of 7,739 patients who were surgically treated for invasive breast cancer at Samsung Medical Center between Dec 1995 and Apr 2013. Luminal A subtype was defined as ER&PR + and HER2- and was further subclassified according to Ki-67 and p53 expression as follows: luminal A (Ki-67-,p53-), luminal A (Ki-67+, p53-), luminal A (Ki-67 -, p53+) and luminal A (Ki-67+, p53+). Low-risk luminal A subtype was defined as negative for both Ki-67 and p53 (luminal A [ki-67-, p53-]), and others subtypes were considered to be high-risk luminal A breast cancer. A cut-off value of 10% for p53 was a good predictor of clinical outcome in all patients and luminal A breast cancer patients. The prognostic role of p53 overexpression for OS and DFS was only significant in luminal A subtype. The combination of p53 and Ki-67 has been shown to have the best predictive power as calculated by the area under curve (AUC), especially for long-term overall survival. In this study, we have shown that overexpression of p53 and Ki-67 could be used to discriminate low-risk luminal A subtype in breast cancer. Therefore, using the combination of p53 and Ki-67 expression in discriminating low-risk luminal A breast cancer may improve the prognostic power and provide the greatest clinical utility.

  5. Use of commercially available rabbit monoclonal antibodies for immunofluorescence double staining

    DEFF Research Database (Denmark)

    Bzorek, M.; Stamp, I.M.; Frederiksen, L.


    Immunohistochemistry, that is, the use of polyclonal and monoclonal antibodies to detect cell and tissue antigens at a microscopical level is a powerful tool for both research and diagnostic purposes. Especially in the field of hematologic disease, there is often a need to detect several antigens...... synchronously, and we report here a fast and easy technique for demonstrating more than 1 antigen in 1 slide using immunofluorescence. We have used commercially available rabbit monoclonal antibodies (Cyclin D1, CD3, CD5, CD23, etc.) paired with mouse monoclonal antibodies (CD7, CD20, CD79a, Pax-5, etc.......) for double immunofluorescence labeling on paraffin-embedded tissue sections. Commercially available rabbit monoclonal antibodies in combination with mouse monoclonal antibodies proved useful in double immunofluorescence labeling on paraffin-embedded tissue, and all combinations used yielded excellent results...

  6. Monoclonal antibodies against human granulocytes and myeloid differentiation antigens. (United States)

    Mannoni, P; Janowska-Wieczorek, A; Turner, A R; McGann, L; Turc, J M


    Monoclonal antibodies (MCA) were obtained by immunizing BALB/c mice with 99% pure granulocytes from normal donors or with a whole leukocyte suspension obtained from a chronic myelogenous leukemia (CML) patient, and then fusing the mouse spleen cells with a 315-43 myeloma cell clone. Four MCA were selected and studied using ELISA, immunofluorescence, cytotoxicity assays, and FACS analysis. Antibodies 80H.1, 80H.3, and 80H.5 (from normals) and 81H.1 (from CML) detected antigens expressed on neutrophils. Antibodies 80H.1 and 80H.3 (IgG) also reacted with monocytes but not with other blood cell subsets. Antibodies 80H.5 and 81H.1 (IgM) were cytotoxic and reacted strongly with most of the cells of the neutrophil maturation sequence, i.e., myeloblasts, promyelocytes, myelocytes, and mature granulocytes. Antibodies 80H.5 and 81H.1 also inhibited CFU-GM growth stimulated by leukocyte feeder layers or placental conditioned media, but did not inhibit BFU-E and CFU-E. Antigens recognized by 80H.3, 80H.5, and 81H.1 were expressed both on a proportion of cells from HL.60, KG.1, ML.1, and K562 myeloid cell lines, and on a proportion of blast cells isolated from patients with acute myelogenous leukemia. They were not found on lymphoid cell lines or lymphoid leukemia cells. These MCA recognize either late differentiation antigens expressed on mature neutrophils and monocytes (80H.1 and 80H.3) or early differentiation antigens (80H.5 and 81H.1) specific to the granulocytic lineage. They may be useful for a better definition of those antigens specific to hematopoietic stem cells and their relationship with normal or neoplastic hematopoiesis.

  7. [ICO-10 monoclonal antibodies to the Thy-1 antigen]. (United States)

    Korotkova, O V; Baryshnikov, A Iu; Tupitsyn, N N; Chimishkian, K L; Kostrykina, V N


    Mouse monoclonal antibodies (MAB) ICO-10 to Thy-1 antigen were obtained. MAB ICO-10 reacted in indirect immunofluorescence test with 5.7 +/- 0.8% human thymocytes. Antibodies did not react with granulocytes, monocytes, T- and non-T cells from peripheral blood, and with marrow cells of healthy donors. MAB ICO-10 reacted with blast cells from 25 of 53 patients with T-cell acute lymphoblastic leukemia (ALL), from 2 of 5 patients with B-cell ALL. This antigen was absent on blood and marrow cells from some patients with ALL, 80 patients with chronic lymphoid leukemia, 54 patients with chronic granulocytic leukemia at the stage of blastic crisis, 128 patients with acute nonlymphoblastic leukemia. Antibodies are specifically bound to thymocytes and spleen cells of Thy 1.1 and Thy 1.2 mice. MAB ICO-10 detect Thy-1 antigen expressed on human hematopoietic cells. MAB ICO-10 may be applied for human leukemia and lymphoma immune diagnosis.

  8. Molecular mechanisms of resistance to the EGFR monoclonal antibody cetuximab. (United States)

    Brand, Toni M; Iida, Mari; Wheeler, Deric L


    The epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase belonging to the HER family of receptor tyrosine kinases. Receptor activation upon ligand binding leads to down stream activation of the PI3K/AKT, RAS/RAF/MEK/ERK and PLCγ/PKC pathways that influence cell proliferation, survival and the metastatic potential of tumor cells. Increased activation by gene amplification, protein overexpression or mutations of the EGFR has been identified as an etiological factor in a number of human epithelial cancers (e.g., NSCLC, CRC, glioblastoma and breast cancer). Therefore, targeting the EGFR has been intensely pursued as a cancer treatment strategy over the last two decades. To date, five EGFR inhibitors, including three small molecule tyrosine kinase inhibitors (TKIs) and two monoclonal antibodies have gained FDA approval for use in oncology. Both approaches to targeting the EGFR have shown clinical promise and the anti-EGFR antibody cetuximab is used to treat HNSCC and CRC. Despite clinical gains arising from use of cetuximab, both intrinsic resistance and the development of acquired resistance are now well recognized. In this review we focus on the biology of the EGFR, the role of EGFR in human cancer, the development of antibody-based anti-EGFR therapies and a summary of their clinical successes. Further, we provide an in depth discussion of described molecular mechanisms of resistance to cetuximab and potential strategies to circumvent this resistance.


    Institute of Scientific and Technical Information of China (English)

    刘小云; 甄永苏


    Objective.Using monoclonal antibody (mAb) Fab′ fragment to develop mAb immunoconjugates for cancer. Methods.Fab′ fragment of mAb 3A5 was prepared by digestion of the antibody with pepsin and then reduced by dithiothreitol (DTT),while Fab′ fragment of mAb 3D6 was obtained by digestion of the antibody with ficin and subsequently reduced by β mercaptoethanol.The conjugation between Fab′ fragment and pingyangmycin (PYM),an antitumor antibiotic,was mediated by dextran T 40.Immunoreactivity of Fab′ PYM conjugates with cancer cells was determined by ELISA,and the cytotoxicity of those conjugates to cancer cells was determined by clonogenic assay.Antitumor effects of the Fab′ PYM conjugates were evaluated by subcutaneously transplanted tumors in mice. Results.The molecular weight of Fab′ fragment was approximately 53 kD,while the average molecular weight of Fab′ PYM conjugate was 170 kD.The Fab′ PYM conjugates showed immunoreactivity with antigen relevant cancer cells and selective cytotoxicity against target cells.Administered intravenously,Fab′ PYM conjugates were more effective against the growth of tumors in mice than free PYM and PYM conjugated with intact mAb. Conclusion.Fab′ PYM conjugate may be capable of targeting cancer cells and effectively inhibiting tumor growth,suggesting its therapeutic potential in cancer treatment.

  10. Effect of polyol sugars on the stabilization of monoclonal antibodies. (United States)

    Nicoud, Lucrèce; Cohrs, Nicholas; Arosio, Paolo; Norrant, Edith; Morbidelli, Massimo


    We investigate the impact of sugars and polyols on the heat-induced aggregation of a model monoclonal antibody whose monomer depletion is rate-limited by protein unfolding. We follow the kinetics of monomer consumption by size exclusion chromatography, and we interpret the results in the frame of two mechanistic schemes describing the enhanced protein stability in the presence of polyols. It is found that the stabilization effect increases with increasing polyol concentration with a comparable trend for all of the tested polyols. However, the stabilization effect at a given polyol concentration is polyol specific. In particular, the stabilization effect increases as a function of polyol size until a plateau is reached above a critical polyol size corresponding to six carbon atoms. Our results show that the stabilization by polyols does not depend solely on the volume fraction filled by the polyol molecules, but is also affected by the polyol chemistry.

  11. Characteristics of Monoclonal Antibody Against Infectious Bursal Disease Virus

    Institute of Scientific and Technical Information of China (English)

    LiYan-Fei; WangWei; 等


    Thirteen strains of monoclonal antibodies(McAbs) against infections bursal disease virus(IBDV) were obtained by using hydridoma technique and their characteristics were studied by double immunodiffusion,enzyme-linked immunosorbent assay(ELISA),virus neutralization test(VNT) and Western-blotting assay (WBA).The result showed that nine of the thirteen McAbs belonged to IgG class and four of them belonged to IgM class.No crossreactions were detected betwween the McAbs and Newscastle disease virus (NDV),infectious bronchitis virus(IBV) and Marek's disease virus(MDV).All of McAbs were positively specific reactive with IBDV and five of them can neutralize viral infectivity.Their recognized epitopes of the neutralizing McAbs were all presented on VP2 of the IBDV.

  12. Characteristics of Monoclonal Antibody Against Infectious Bursal Disease Virus

    Institute of Scientific and Technical Information of China (English)


    Thirteen strains of monoclonal antibodies (McAbs) against infectious bursal disease virus (IBDV) were obtained by using hybridoma technique and their characteristics were studied by double immunodiffusion,en- zyme- linked immunosorbent assay (ELISA), virus neutralization test (VNT) and Western- blotting assay (WBA). The result showed that nine of the thirteen McAbs belonged to IgG class and four of them belonged to IgM class. No crossreactions were detected betwween the McAbs and Newscastle disease virus (NDV) ,in- fectious bronchitis virus(IBV) and Marek's disease virus(MDV). All of McAbs were positively specific reac- tive with IBDV and five of them can neutralize viral infectivity. Their recognized epitopes of the neutralizing McAbs were all presented on VP2 of the IBDV.

  13. Preparation and Identification of Monoclonal Antibodies Against Vibrio anguillarum

    Institute of Scientific and Technical Information of China (English)

    Chen Shiyong(陈师勇); Zhang Peijun; Mo Zhaolan; Zhang Zhendong; Zou Yuxia; Xu Yongli


    Monoclonal antibodies (Mabs) against V.anguillarum strain M3 are prepared, and their isotypes are also characterized. Among them, C1C5 is the only Mab which does not crossreact with other eleven non-V.anguillarum strains. The proteinase K digestion test shows that the epitopes recognized by C1C5, C6C3 and C6C32 Mabs contained protein. The periodate oxidation test showed that the epitopes recognized by Mabs except C1C5 are glycosylated. In addition, results of additivity test indicate that the epitopes recognized by C6C3 and C6C32 Mabs are similar, and quite different from those recognized by Mab C1C5.

  14. [Monoclonal antibodies for the treatment of multiple sclerosis]. (United States)

    Sánchez-Seco, Victoria Galán; Casanova Peño, Ignacio; Arroyo González, Rafael


    Until the mid 1990s, with the appearance of interferon beta and glatiramer acetate, there was no treatment for multiple sclerosis (MS). However, due to their moderate therapeutic potential in some patients, a broad search was continued to find new and more effective treatment strategies, largely concentrated on monoclonal antibodies (MOAB). Natalizumab, the first MOAB for the treatment of MS, was approved at the end of 2004, representing a major advance in the field of neuroimmunology. Today, there is broad experience with natalizumab and other MOAB (alemtuzumab, daclizumab, rituximab, ocrelizumab, ofatumumab and anti-lingo-1) that are pending commercialization or are under phase II or III of development with promising results. The present review analyzes the efficacy and safety results of all these drugs. Copyright © 2014 Elsevier España, S.L.U. All rights reserved.

  15. Pharmacokinetic, pharmacodynamic and immunogenicity comparability assessment strategies for monoclonal antibodies. (United States)

    Putnam, Wendy S; Prabhu, Saileta; Zheng, Yanan; Subramanyam, Meena; Wang, Yow-Ming C


    Regulatory guidance stipulates that comparability assessment is required to support manufacturing process changes during the development of a biological product or post-approval. However, strategies for assessing the comparability of pre- and post-change materials are still evolving. A hierarchical risk-based approach is recommended, starting with analytical testing to ensure quality, followed by biological characterization and, if needed, in vivo pharmacokinetic (PK), PK-pharmacodynamic (PD), safety and/or efficacy studies. The need for an in vivo study and the type of study required depend on the magnitude and the potential impact of the changes and the timing in the development process. This review discusses factors affecting the PK, PD and immunogenicity of monoclonal antibodies, and provides guidance for determining non-clinical and clinical comparability assessment strategies. Copyright © 2010 Elsevier Ltd. All rights reserved.

  16. Selection of Ceratitis capitata (Diptera: Tephritidae) specific recombinant monoclonal phage display antibodies for prey detection analysis. (United States)

    Monzó, César; Urbaneja, Alberto; Ximénez-Embún, Miguel; García-Fernández, Julia; García, José Luis; Castañera, Pedro


    Several recombinant antibodies against the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), one of the most important pests in agriculture worldwide, were selected for the first time from a commercial phage display library of human scFv antibodies. The specificity and sensitivity of the selected recombinant antibodies were compared with that of a rabbit polyclonal serum raised in parallel using a wide range of arthropod species as controls. The selected recombinant monoclonal antibodies had a similar or greater specificity when compared with classical monoclonal antibodies. The selected recombinant antibodies were successfully used to detect the target antigen in the gut of predators and the scFv antibodies were sequenced and compared. These results demonstrate the potential for recombinant scFv antibodies to be used as an alternative to the classical monoclonal antibodies or even molecular probes in the post-mortem analysis studies of generalist predators.

  17. PCSK9 Inhibition With Monoclonal Antibodies: Modern Management of Hypercholesterolemia. (United States)

    Ito, Matthew K; Santos, Raul D


    Current guidelines for hypercholesterolemia treatment emphasize lifestyle modification and lipid-modifying therapy to reduce the risk for cardiovascular disease. Statins are the primary class of agents used for the treatment of hypercholesterolemia. Although statins are effective for many patients, they fail to achieve optimal reduction in lipids for some patients, including those who have or are at high risk for cardiovascular disease. The PCSK9 gene was identified in the past decade as a potential therapeutic target for the management of patients with hypercholesterolemia. Pharmacologic interventions to decrease PCSK9 levels are in development, with the most promising approach using monoclonal antibodies that bind to PCSK9 in the plasma. Two monoclonal antibodies, alirocumab and evolocumab, have recently been approved for the treatment of hypercholesterolemia, and a third one, bococizumab, is in phase 3 clinical development. All 3 agents achieve significant reductions in levels of low-density lipoprotein cholesterol, as well as reductions in non-high-density lipoprotein cholesterol, apolipoprotein B, and lipoprotein(a). Long-term outcome trials are under way to determine the sustained efficacy, safety, and tolerability of PCSK9 inhibitors and whether this novel class of agents decreases the risk for major cardiovascular events in patients on lipid-modifying therapy. Available data suggest that PCSK9 inhibitors provide a robust reduction in atherogenic cholesterol levels with a good safety profile, especially for patients who fail to obtain an optimal clinical response to statin therapy, those who are statin intolerant or have contraindications to statin therapy, and those with familial hypercholesterolemia. © 2016, The Authors. The Journal of Clinical Pharmacology Published by Wiley Periodicals, Inc. on behalf of American College of Clinical Pharmacology.

  18. Evaluation of an Optimal Cut-Off Point for the Ki-67 Index as a Prognostic Factor in Primary Breast Cancer: A Retrospective Study (United States)

    Tashima, Rumiko; Nishimura, Reiki; Osako, Tomofumi; Nishiyama, Yasuyuki; Okumura, Yasuhiro; Nakano, Masahiro; Fujisue, Mamiko; Toyozumi, Yasuo; Arima, Nobuyuki


    The Ki-67 index is an important biomarker for indicating the proliferation of cancer cells and is considered to be an effective prognostic factor for breast cancer. However, a standard cut-off point for the Ki-67 index has not yet been established. Therefore, the aim of this retrospective study was to determine an optimal cut-off point in order to establish it as a more accurate prognostic factor. Immunohistochemical analysis of the Ki-67 index was performed on 4329 patients with primary breast cancer from August 1987 to March 2012. Out of this sample, there were 3186 consecutive cases from September 1997 with simultaneous evaluations of ER, PgR and HER2 status. Cox's proportional hazard model was used to perform univariate and multivariate analyses of the factors related to OS. The hazard ratios (HR) and the p values were then compared to determine the optimal cut-off point for the Ki-67 index. The median Ki-67 index value was 20.5% (mean value 26.2%). The univariate analysis revealed that there was a statistically significant negative correlation with DFS and OS and the multivariate analysis revealed that the Ki-67 index value was a significant factor for DFS and OS. The top seven cut-off points were then carefully chosen based on the results of the univariate analysis using the lowest p-values and the highest HR as the main selection criteria. The multivariate analysis of the factors for OS showed that the cut-off point of 20% had the highest HR in all of the cases. However, the cutoff point of 20% was only a significant factor for OS in the Luminal/HER2- subtype. There was no correlation between the Ki-67 index value and OS in any of the other subtypes. These data indicate that the optimal cut-off point of 20% is the most effective prognostic factor for Luminal/HER2- breast cancer. PMID:26177501


    Directory of Open Access Journals (Sweden)

    Rakheja Mahima, Singh Pratyush, Shergill Ankur K, Guddattu Vasudeva,Solomon Monica C


    Full Text Available Background: Cancer cells exhibit a characteristic intrinsic ability of uncontrolled cell proliferation which plays a vital role in tumor development and progression. The Ki-67 antigen expression is strictly associated with cell proliferation which detects the cells entering the cell cycle. In addition, computer aided image analysis provides an objective and highly reproducible histological evaluation of OSCC. Assessment of Ki-67 expression along with morphometry may help in early and precise diagnosis and prognostication of OSCC. Aims: To correlate the ki-67 expression and the morphometrical parameters of oral squamous cell carcinomas and to assess the efficacy of the correlation in OSCC prognostication. Methods and material: OSCC cases (n=105 were examined immuno histochemically using Ki-67 antigen The nuclear area (NA, cell area (CA, nuclear perimeter (NP and cell perimeter (CP assessed using “image J 1.34 software”. The data was statistically analyzed. Results: Highly significant correlation was found between Ki-67 expression and the advancing grades of OSCC (p<.0001. NA (p=0.025, CA (p<.0001, NP (p=.027 and CP (p<.0001 of poorly differentiated OSCC were significantly lower than well differentiated and moderately differentiated OSCC. Follow up analysis revealed nuclear area and cell area to be higher in the recurrent cases than the non recurrent ones. Conclusion: Assessment of Ki-67 expression and morphometry help in early and precise diagnosis and prognostication of OSCC. A correlation between Ki-67 expression and morphometrical analysis could not be ascertained in this study. Further studies with a larger sample number may provide more definitive results.

  20. Correlation of (18)F-FDG uptake on PET/CT with Ki67 immunohistochemistry in pre-treatment epithelial ovarian cancer. (United States)

    Mayoral, M; Paredes, P; Saco, A; Fusté, P; Perlaza, P; Tapias, A; Fernandez-Martinez, A; Vidal, L; Ordi, J; Pavia, J; Martinez-Roman, S; Lomeña, F


    Standardized uptake value (SUV) and volumetric parameters such as metabolic tumour volume (MTV) and total lesion glycolysis (TLG) from (18)F-FDG PET/CT are useful criteria for disease prognosis in pre-operative and post-treatment epithelial ovarian cancer (EOC). Ki67 is another prognostic biomarker in EOC, associated with tumour aggressiveness. The aim of this study is to evaluate the association between (18)F-FDG PET/CT measurements and Ki67 in pre-treatment EOC to determine if PET/CT parameters could non-invasively predict tumour aggressiveness. A pre-treatment PET/CT was performed on 18 patients with suspected or newly diagnosed EOC. Maximum SUV (SUVmax), mean SUV (SUVmean), whole-body MTV (wbMTV), and whole-body TLG (wbTLG) with a threshold of 30% and 40% of the SUVmax were obtained. Furthermore, Ki67 index (mean and hotspot) was estimated in tumour tissue specimens. Immunohistochemical findings were correlated with PET parameters. The mean age was 57.0 years old (standard deviation 13.6 years). A moderate correlation was observed between mean Ki67 index and SUVmax (r=0.392), SUVmean 30% (r=0.437), and SUVmean 40% (r=0.443), and also between hotspot Ki67 index and SUVmax (r=0.360), SUVmean 30% (r=0.362) and SUVmean 40% (r=0.319). There was a weaker correlation, which was inversely negative, between mean and hotspot Ki67 and volumetric PET parameters. However, no statistical significant differences were found for any correlations. SUVmax and SUVmean were moderately correlated with Ki67 index, whereas volumetric PET parameters overall showed a weaker correlation. Thus, SUVmax and SUVmean could be used to assess tumour aggressiveness in pre-treatment EOC. Copyright © 2017 Elsevier España, S.L.U. y SEMNIM. All rights reserved.

  1. Correlations of {sup 18}F-fluorothymidine uptake with pathological tumour size, Ki-67 and thymidine kinase 1 expressions in primary and metastatic lymph node colorectal cancer foci

    Energy Technology Data Exchange (ETDEWEB)

    Nakajo, Masatoyo; Nakajo, Masayuki [Kagoshima University, Department of Radiology, Graduate School of Medical and Dental Sciences, Kagoshima (Japan); Nanpuh Hospital, Department of Radiology, Kagoshima (Japan); Kajiya, Yoriko; Tani, Atsushi [Nanpuh Hospital, Department of Radiology, Kagoshima (Japan); Goto, Yuko; Higashi, Michiyo [Kagoshima University, Department of Human Pathology, Graduate School of Medical and Dental Sciences, Kagoshima, 890-8544 (Japan); Jinguji, Megumi; Fukukura, Yoshihiko [Kagoshima University, Department of Radiology, Graduate School of Medical and Dental Sciences, Kagoshima (Japan); Tanaka, Sadao [Nanpuh Hospital, Department of Pathology, Kagoshima (Japan)


    To examine correlations of {sup 18}F-fluorothymidine (FLT) uptake with pathological tumour size and immunohistochemical Ki-67, and thymidine kinase 1 (TK-1) expressions in primary and metastatic node colorectal cancer foci. Thirty primary cancers (PCs) and 37 metastatic nodes (MNs) were included. FLT uptake was assessed by visual scores (non-visible: 0-1 and visible: 2-4), standardized uptake value (SUV), and correlated with size, Ki-67, and TK-1. SUV was measured in visible lesions. FLT heterogeneity was assessed by visual scores (no heterogeneous uptake: 0 and heterogeneous uptake: 1-4). Forty-two lesions were visible. The visible group showed significantly higher values than the non-visible group in size, Ki-67, and TK-1 (each p < 0.05). Size correlated significantly with visual score (PC; ρ = 0.74 and MN; ρ = 0.63), SUVmax (PC; ρ = 0.49, and MN; ρ = 0.76), and SUVmean (PC; ρ = 0.40 and MN; ρ = 0.76) (each p < 0.05). Visual score correlated significantly with size (ρ = 0.86), Ki-67max (ρ = 0.35), Ki-67mean (ρ = 0.38), TK-1max (ρ = 0.35) and TK-1mean (ρ = 0.25) (each p < 0.05). No significant correlations were found between FLT uptake and Ki-67 or TK-1 in 42 visible lesions (each p > 0.05). Heterogeneous FLT uptake was noted in 73 % (22/30) of PCs. FLT uptake correlated with size. Heterogeneous FLT distribution in colorectal cancers may be one of the causes of weak or lack of FLT uptake/Ki-67 or TK-1 correlation. (orig.)

  2. Changes of expression of estrogen and progestrone receptors, human epithelial growth factor receptor 2 and Ki-67 after neoadjuvant chemotherapy in the treatment of breast cancer. (United States)

    Li, M L; Dong, Y; Luan, S L; Zhao, Z H; Ning, F L


    Recent studies suggest that the development and prognosis of breast cancer is in close correlation to molecular subtype of breast cancer. Neoadjuvant chemotherapy has been extensively applied in the treatment of local breast cancer in advanced stage. In order to verify the correlation between expression changes of estrogen receptor, progestrone receptor, human epithelial growth factor receptor 2 and Ki-67 after neoadjuvant chemotherapy and neoadjuvant chemotherapy, we studied 120 patients with stage IIAIIIC breast cancer who underwent neoadjuvant chemotherapy in Binzhou Medical University Hospital, Shandong, China from February 2011 to February 2015. Clinical characteristics were retrospectively analyzed. The expression of estrogen receptor, progesterone receptor, human epithelial growth factor receptor 2 and Ki-67 of patients were detected using the immunohistochemical method before and after neoadjuvant chemotherapy. The results suggest that the overall remission rate of neoadjuvant chemotherapy was 76.7% (92/120) of which 16.7% (20/120) of cases had complete remission, 60% (72/120) had partial remission and 23.3% (28/120) were stable. There were no cases of progressive disease. The property of estrogen receptor and the expression of Ki-67 of primary tumor were correlated to the remission rate of neoadjuvant chemotherapy (P less than 0.05). The expression of Ki-67 had a significant decline after neoadjuvant chemotherapy, and the difference had statistical significance (P less than 0.05). The difference in expression of estrogen receptor, progesterone receptor and human epithelial growth factor receptor 2 before and after neoadjuvant chemotherapy had statistical significance (P > 0.05). Hence, it can be concluded that breast cancer patients with negative estrogen receptor expression and high Ki-67 expression before neoadjuvant chemotherapy can achieve better curative effects. Neoadjuvant chemotherapy cannot change the expression states of estrogen receptor

  3. Predictive value of Ki67 and p53 in locally advanced rectal cancer: Correlation with thymidylate synthase and histopathological tumor regression after neoadjuvant 5-FU-based chemoradiotherapy

    Institute of Scientific and Technical Information of China (English)

    Christiane Jakob; Torsten Liersch; Wolfdietrich Meyer; Heinz Becker; Gustavo B Bare; Daniela E Aust


    AIM:To investigate the predictive value of Ki67 and p53 and their correlation with thymidylate synthase(TS) gene expression in a rectal cancer patient cohort treated according to a standardized recommended neoadjuvant treatment regimen.METHODS:Formalin fixed,paraffin embedded pretherapeutical tumor biopsies (n=22) and posttherapeutical resection specimens(n=40)from patients with rectal adenocarcinoma (clinical UICC stage Ⅱ/Ⅲ)receiving standardized neoadjuvant 5-fiuorouracil(5-FU)based chemoradiotherapy were studied for Ki67 and p53 expression by immunohistochemistry and correlated with TS mRNA expression by quantitative TaqMan realtime PCR after laser microdissection.The results were compared with histopathological tumor regression according to a standardized semiquantitative score grading system.RESULTS:Responders(patients with high tumor regression)showed a significantly lower Ki67 expression than non-responders in the pre-therapeutical tumor biopsies (81.2% vs16.7%;P<0.05) as well as in the post-therapeutical resection specimens (75.8%vs14.3%;P<0.01).High TS mRNA expression was significantly correlated with a high Ki67 index and low TS mRNA expression was significantly correlated with a low Ki67 index in the pre-therapeutical tumor biopsies (corr.coef.=0.46;P<0.01)as well as in the posttherapeutical resection specimens (corr.coef.=0.40;P<0.05).No significant association was found between p53 and TS mRNA expression or tumor regression.CONCLUSION:Ki67 has,like TS,predictive value in rectal cancer patients after neoadjuvant 5-FU based chemoradiotherapy.The close correlation between Ki67 and TS indicates that TS is involved in active cell cycle processes.

  4. The abnormal expre ssion of retinoic acid receptor-β, p53 and Ki67 protein in normal, premalignant and malignant esophageal tissues

    Institute of Scientific and Technical Information of China (English)

    Min Xu; Yu-Lan Jin; Jun Fu; Hong Huang; Sheng-Zu Chen; Ping Qu; Hai-Mei Tian; Zhao-Yang Liu; Wei Zhang


    AIM: Esophageal cancer remains a significant healthiproblem worldwide. It is important to investigate alterationsin expression of retinoic acid receptor-β, p53 and Ki67proteins in esophageal carcinogenesis.METHODS: To find biomarkers for early identification ofesophageal cancer, we analyzed the retinoic acid receptor-j3,p53 protein and the proliferation marker Ki67 in surgicalspecimens of normal, mildly, and severely dysplastic andmalignant esophageal tissues by in situ hybridization ofRNA and immunohistochemistry.RESULTS: RAR-β was expressed in 94.3 %(33/35) of normalmucosae, 67.8 %(19/28) of the mild, 58.1% (18/31) of thesevere ieaions and 53.2 % (116/218) of tumor samples. RAR-β mRNA was expressed in 62.7 % (42/67), 55.1% (43/78) and29.2 % (7/24) of well, moderated and poorly differentiatedSSCs. The p53 and Ki67 proteins were 5.9 % (2/34) of thenormal mucosa. P53 and Ki67 stained positively in 10.7 % (3/28) and 21.4 % (6/28) of mild dysplasia, and 51.6 %(16/31)and 58. 1% (18/31) of severely dysplasia respectively.Samples from esophageal cancer showed no higher levers ofp53 and Ki67 expression than seen in severely dysplasticlesions. There was significant difference of RAR-β、 p53 andKi67 expression between normal mucosa and dysplatictissue or esophageal cancer.CONCLUSION: Loss of RAR-β expression and acctnulation ofp53 and Ki67 proteins may serve as biomarkers for earlyidentification of esophageal cancer in the high-riskpopulations.

  5. Evaluation of Telomerase (hTert, Ki67 and p16ink4a expressions in low and high-grade cervical intraepithelial lesions

    Directory of Open Access Journals (Sweden)


    Full Text Available ABSTRACT Objective : to study the association between the histological grading of cervical intraepithelial neoplasia (CIN I, CIN II and CIN III and the immunohistochemical expression for p16ink4a, hTert and Ki67, as well as to evaluate the relationship of these markers with the risk of recurrence after surgical treatment. Methods : we studied a historical cohort of 94 women with intraepithelial lesions CIN I (low grade, CIN II and CIN III (high grades submitted to conization or electrosurgical excision of the transformation zone. We evaluated all surgical specimens for immunohistochemical expression of p16ink4a, hTert and Ki67. Results : the mean age was 38.2 years; p16ink4a was absent in most CIN I cases. In patients with CIN II or I/II (association of low and high-grade lesions, we observed p16ink4a ≤10%. In patients with CIN III, we found a higher expression frequency of p16ink4a >50%. In CIN I, the majority had Ki67≤10% and low frequency of Ki67>50%. In the CIN III category, there were fewer patients with Ki67≤10%, and Ki67 was absent in most patients of CIN II and III groups. There was no association between hTert expression and histologic grade. There were no statistically significant differences between the expression of the markers in patients with and without recurrence. Conclusion : there was a statistically significant association of p16ink4a and Ki67 with histological grade. The markers' expression, as for disease recurrence, was not statistically significant in the period evaluated.

  6. The expression of p53,Ki-67 in colorectal adenocarcinoma with neuroen-docrine differentiation and the correlation between the expression and its prognosis%伴神经内分泌分化结直肠腺癌中p53、Ki-67的表达及其与预后的关系

    Institute of Scientific and Technical Information of China (English)

    张海勇; 郦秀芳; 吕艳婷; 胡孟钧


    Objective To investigate the expression of p53, Ki-67 in colorectal adenocarcinoma with neuroendocrine differentiation (NED), and to explore the correlation between the expression and its prognosis. Methods The expression of NSE, CgA, Syn, CD-57, p53, Ki-67 in 65 colorectal adenocarcinoma were determined by using immunohistochemi-cal staining. According the neuroendocrine index, 65 cases were divided into two groups,NED positive group(35 cases) and NED negative group (30 cases),and analyzed the influence of erpression of p53, ki-67cm prognosis. Results The positive expression rates of p53, Ki-67 in NED positive group were higher than those in NED negative group (P<0.05). The expression of p53,Ki-67 in NED positive group were significantly associated with lymph node metastasis, differen-tiation and DUCK stages (P<0.05). The 5-year survival rate of NED negative group was obviously higher than that of NED positive group (P<0.05);In NED positive group, the 5-year survival rate of p53 negative and Ki-67 negative ex-pression were significantly higher than that of p53 positive and Ki-67 positive expression (P<0.05). Using Cox regres-sion, lymph node metastasis, differentiation, DUCK stages and the expression of p53,Ki-67 were strong independent prognostic factors for five years of disease-free survival rate of colorectal adenocarcinoma with NED patients (P<0.05). Conclusion The high expression of p53, Ki-67 may be associated with colorectal adenocarcinoma with NED and play an important role in malignant proliferation. p53,Ki-67 are valuable prognostic factors for colorectal adenocarcinoma with NED patients.%目的:探讨伴神经内分泌分化(NED)结直肠腺癌中p53、Ki-67的表达及其与预后的关系。方法采用SP法检测NSE、CgA、Syn、CD-57及p53、Ki-67蛋白在65例结直肠腺癌组织中的表达,分为NED阳性组35例和NED阴性组30例,并分析p53、Ki-67表达对预后的影响。结果 NED阳性组中p53、Ki-67阳性表达

  7. Antibodies to poliovirus detected by immunoradiometric assay with a monoclonal antibody

    Energy Technology Data Exchange (ETDEWEB)

    Spitz, M.; Fossati, C.A.; Schild, G.C.; Spitz, L.; Brasher, M. (National Inst. for Biological Standards and Control, London (UK))


    An immunoradiometric assay (IRMA) for the assay of antibodies to poliovirus antigens is described. Dilutions of the test sera or whole (finger prick) blood samples were incubated with the poliovirus antigen bound to a solid phase and the specific antibody was detected by the addition of a mouse anti-human IgG monoclonal antibody (McAb), which was itself revealed by iodinated sheep IgG antimouse F(ab). The authors have shown that this technique is suitable for the estimation of IgG anti-poliovirus antibodies induced in children following polio vaccine. The present study shows that SPRIA provides a simple and inexpensive method for serological studies with poliovirus particularly for use in large-scale surveys.

  8. Ki-67对三阴性乳腺癌新辅助化疗敏感性预测作用的Meta分析%Predict function of Ki-67 to chemosensitivity of neoadjuvant chemotherapy in triple-negative breast cancer : a meta-analysis

    Institute of Scientific and Technical Information of China (English)

    章国晶; 刘兆喆; 郭放; 谢晓冬


    目的:评价新辅助化疗中Ki-67的表达情况对三阴性乳腺癌(TNBC)获得病理完全缓解(pCR)的预测作用.方法:通过互联网检索相关文献,应用Review Manager软件对数据进行处理,系统评价Ki-67表达情况与TNBC在新辅助化疗中获得pCR的关系.结果:Ki 67基因高表达的TNBC患者pCR率明显高于Ki-67基因低表达的TNBC患者,总体合并优势比(OR)为7.35,95%CI:2.39~22.60,文献之间无明显异质性,无明显发表偏倚.结论:Ki-67可作为新辅助化疗中预测TNBC化疗敏感性的标志.%OBJECTIVE; The meta-analysis was conducted to estimate the predict function of Ki-67 to pathological complete response (pCR) rates of neoadjuvant chemotherapy in triple-negative breast cancer(TNBC). METHODS: Relevant literatures were selected by searching the internet. Review manager software was used to merge extractive data. RESULTS: The pCR rates were much higher in Ki-67 high expression TNBC than that in Ki-67 low expression TNBC. The merged odds ratio(OR) was 7. 35(95% confidence interval: 2. 39 - 22. 60). No obvious statistical heterogeneity and publication bias were detected. CONCLUSION: Ki-67 was a predictor to chemosensitivity of neoadjuvant chemotherapy in TNBC.

  9. Cutaneous hybrid tumor composed of epidermal cyst and cystic pilomatricoma expressing p53 and high Ki-67 labeling. (United States)

    Terada, Tadashi


    Hybrid tumor composed of epidermal cyst and pilomatricoma has been reported in only four times in the English literature. Herein, a cutaneous hybrid tumor composed of epidermal cyst and cystic pilomatricoma in a 58-year-old woman was presented. The tumor was located in the scalp, and measured 1 x 1 x 1 cm. The tumor was cystic and contained atheromatous materials. Histologically, the cyst was composed of epidermal cyst (50% in area) and pilomatricoma (50% in area). The pilomatricoma was composed mostly of basophilic cells. Interestingly, the pilomatricoma element of the cyst showed immunoreactive p53 and Ki-67 (labeling=40%). In conclusion, the fifth case of hybrid tumor composed of epidermal cyst and pilomatricoma was presented.

  10. Preparation and Biological Evaluation of 188Re Labeled Monoclonal Antibody TGLA

    Institute of Scientific and Technical Information of China (English)

    WEN; Kai; ZHANG; Jun-li; CHEN; Bao-jun; CUI; Hai-ping


    <正>Monoclonal antibody TGLA is a specific targeting CD20 chimeric antibody. It can kill tumor cells and inhibit tumor cells’ growth effectively, which has been applied to clinical therapy of lymphoma cell B. 188 Re is easy to get, and emits both β and γ rays. 188Re labeled monoclonal antibody TGLA can be used for the study of lymphoma therapy and imaging. This work got the product 188Re-TGLA by direct labeling

  11. Expression of Ki67, BCL-2, and COX-2 in canine cutaneous mast cell tumors: association with grading and prognosis. (United States)

    Vascellari, M; Giantin, M; Capello, K; Carminato, A; Morello, E M; Vercelli, A; Granato, A; Buracco, P; Dacasto, M; Mutinelli, F


    The expression of Ki67, BCL-2, and COX-2 was investigated in 53 canine cutaneous mast cell tumors (MCTs) by immunohistochemistry and quantitative real time polymerase chain reaction (qPCR) to evaluate their prognostic significance and the association with the histologic grading and the mitotic index (MI). MCTs were graded according to the Patnaik grading system and the novel 2-tier grading system proposed by Kiupel. The numbers of mitotic figures/10 high-power fields (MI) were counted. Both grading systems were significantly associated with prognosis. The Patnaik grading was of limited prognostic value for grade 2 MCTs, with 23% being associated with mortality. The concordance among pathologists was strongly improved by the application of the 2-tier grading system, and 71% of high-grade MCTs were associated with a high mortality rate. MI and Ki67 protein expression were significantly associated with grading and survival. No significant association between BCL-2 protein expression and either grading system or health status was observed. BCL-2 mRNA expression was significantly higher in grade 2 than in grade 1 MCTs, while no statistically significant differences were detected between low- and high-grade MCTs. The increased BCL-2 mRNA level was significantly associated with increased mortality rate. The COX-2 protein expression was detected in 78% of the MCTs investigated. However, neither association with the tumor grade nor with the health status was observed. COX-2 mRNA was significantly up-regulated in MCTs compared to surgical margins and control skin tissue, but it was neither associated with tumor grade nor with survival.

  12. Expression of Ki-67, Oct-4, γ-tubulin and α-tubulin in human tooth development. (United States)

    Kero, Darko; Novakovic, Josip; Vukojevic, Katarina; Petricevic, Josko; Kalibovic Govorko, Danijela; Biocina-Lukenda, Dolores; Saraga-Babic, Mirna


    To analyze factors controlling cell proliferation and differentiation, and appearance of primary cilia during the cap and bell stages of incisor or/and canine human enamel organs. Qualitative and quantitative analysis of proliferating Ki-67 positive cells and expression of γ-tubulin, α-tubulin and Oct-4 was immunohistochemically analyzed in the cap an bell stages of 10 developing human incisor and canine germs, 8-21 weeks old. During the analyzed period, ratio of Ki-67 positive cells changed in outer enamel epithelium from 48.86% to 24.52%, in inner enamel epithelium increased from 56.11% to 60.06% and then dropped to 44.24%. While in dental papilla proliferation first increased from 46.26% to 55.45%, and then dropped to 22.08%, a constant decrease of proliferation characterized enamel reticulum (from 46.26% to 15.49%). Strong cytoplasmic Oct-4 expression characterized epithelial parts of enamel organ, particularly the differentiating ameloblasts. During further development, Oct-4 expression shifted to both nuclear and cytoplasmic expression in mesenchymal tooth components. Primary cilia characterized most of the cells in developing enamel organ. While non-ciliated (proliferating) cells mainly contained two centrioles (γ-tubulin), the primary cilia (α-tubulin) were arising from basal bodies (γ-tubulin) of non-proliferating cells. We suggest that increase in cell proliferation enables growth of enamel organ, while its selective decrease leads to disintegration of some tooth parts. Drop of proliferation coincided with initiation of ameloblast and odontoblast differentiation. Additionally, cell differentiation was accompanied by increased expression of Oct-4 and probably by signalling via primary cilia, both regulating processes of cell proliferation and differentiation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  13. [Sorting role of p16(INK4a)/Ki-67 double immunostaining in the cervical cytology specimens of ASCUS and LSIL cases]. (United States)

    Yu, J; Zhu, H T; Zhao, J J; Su, J Z; Xia, Y D


    Objective: To investigate the sorting effect of p16(INK4a)/Ki-67 double immunostaining method in patients with atypical squamous cells of undetermined significance (ASCUS) or low-grade squamous intraepithelial lesion (LSIL) cytology results. Methods: Four-hundred and twenty cases collected during April 2014 to February 2015 of cervical cytology of ASCUS (n=318) and LSIL (n=102) were selected, and residual liquid-based cytology specimens were used for p16(INK4a)/Ki-67 double immunostaining. The sensitivity and specificity of the detection of cervical precancerous lesions and cervical cancer were calculated, and the results were compared with high risk HPV. Taking histological follow-up as the gold standard, the test was considered positive when at least one cell exhibited p16(INK4a)/Ki-67 co-staining, without requirement of adjunct morphologic interpretation of positive cells. Results: Further screening CIN2+ in cytology ASCUS and LSIL group , the sensitivity of p16(INK4a)/Ki-67 double immunostaining was slightly lower than high risk HPV (84.2% vs. 94.7%), while the specificity was higher (84.0% vs. 53.9%). For ASCUS patients, the sensitivity of p16(INK4a)/Ki-67 double immunostaining and high risk HPV was 82.6% and 91.3%, and the specificity was 88.8% and 63.7%, respectively. For LSIL patients, the sensitivity of p16(INK4a)/Ki-67 double immunostaining and high risk HPV was 86.7% and 100.0%, and the specificity was 67.8% and 20.7%, respectively. For patients younger and older than 30 years, specificity of p16(INK4a)/Ki-67 double immunostaining was both higher than that of high risk HPV (80.8% vs. 42.3%; 84.6% vs. 56.9%). Conclusions: p16(INK4a)/Ki-67 double immunostaining can effectively identify the high risk population in ASCUS or LSIL, with higher specificity than high risk HPV test. p16(INK4a)/Ki-67 double immunostaining may benefit patients younger than 30 years of age as a preliminary or potential cytology-combining screening tool.

  14. Prognostic impact of proliferation for resected early stage 'pure' invasive lobular breast cancer: Cut-off analysis of Ki67 according to histology and clinical validation. (United States)

    Carbognin, Luisa; Sperduti, Isabella; Fabi, Alessandra; Dieci, Maria Vittoria; Kadrija, Dzenete; Griguolo, Gaia; Pilotto, Sara; Guarneri, Valentina; Zampiva, Ilaria; Brunelli, Matteo; Orvieto, Enrico; Nortilli, Rolando; Fiorio, Elena; Parolin, Veronica; Manfrin, Erminia; Caliò, Anna; Nisticò, Cecilia; Pellini, Francesca; Scarpa, Aldo; Pollini, Giovanni Paolo; Conte, Pierfranco; Tortora, Giampaolo; Bria, Emilio


    The intent of this analysis was to investigate and validate the prognostic potential of Ki67 in a multi-center series of patients affected by early stage 'pure' invasive lobular carcinoma (ILC). Clinical-pathological data of patients affected by ILC were correlated with overall survival and disease-free survival (OS/DFS); data from a parallel invasive ductal carcinoma (IDC) patients' cohort were gathered as well. The maximally selected Log-Rank statistics analysis was applied to Ki67 continuous variable to estimate the appropriate cut-off. The Subpopulation Treatment Effect Pattern Plot (STEPP) analysis was performed as well. Data from overall 1097 (457/222 ILC: training/validation set; 418 IDC) patients were gathered. The identified optimal Ki67 cut-offs were 4% and 14% for DFS in ILC and IDC cohort, respectively. In ILC patients, the Ki67 cut-off was an independent OS predictor. Ten-years OS and DFS were 89.9% and 77.2% (p = 0.007) and 79.4% and 69.2% (p = 0.03) for patients with Ki67 ≤ 4% and >4%, respectively. In IDC patients, 10-years OS was 93.8% and 71.7%, p = 0.02, DFS was 84.0% and 52.6%, p = 0.0003, for patients with Ki67 ≤ 14% and >14%, respectively. In the validation set, the optimal Ki67 OS cut-off was 5%. The STEPP analysis showed that in the presence of low Ki67 values, IDC patients have a better DFS than ILC patients, while with the increase of values the prognosis tends to overlap. Despite the retrospective design of the study, the prognostic relevance of Ki67 (as well as its optimal cut-off) seems to significantly differ according to breast cancer histology. Copyright © 2017 Elsevier Ltd. All rights reserved.


    NARCIS (Netherlands)



    The selectivity of anticancer agents may be improved by antibody-directed enzyme prodrug therapy (ADEPT), The immunogenicity of antibody-enzyme conjugates and the low tumor to normal tissue ratio calls for the use of a human enzyme and the development of a monoclonal antibody (MAb) against that enzy

  16. Profiling formulated monoclonal antibodies by (1)H NMR spectroscopy. (United States)

    Poppe, Leszek; Jordan, John B; Lawson, Ken; Jerums, Matthew; Apostol, Izydor; Schnier, Paul D


    Nuclear magnetic resonance (NMR) is arguably the most direct methodology for characterizing the higher-order structure of proteins in solution. Structural characterization of proteins by NMR typically utilizes heteronuclear experiments. However, for formulated monoclonal antibody (mAb) therapeutics, the use of these approaches is not currently tenable due to the requirements of isotope labeling, the large size of the proteins, and the restraints imposed by various formulations. Here, we present a new strategy to characterize formulated mAbs using (1)H NMR. This method, based on the pulsed field gradient stimulated echo (PGSTE) experiment, facilitates the use of (1)H NMR to generate highly resolved spectra of intact mAbs in their formulation buffers. This method of data acquisition, along with postacquisition signal processing, allows the generation of structural and hydrodynamic profiles of antibodies. We demonstrate how variation of the PGSTE pulse sequence parameters allows proton relaxation rates and relative diffusion coefficients to be obtained in a simple fashion. This new methodology can be used as a robust way to compare and characterize mAb therapeutics.

  17. Analysis of viral clearance unit operations for monoclonal antibodies. (United States)

    Miesegaes, George; Lute, Scott; Brorson, Kurt


    Demonstration of viral clearance is a critical step in assuring the safety of biotechnology products. We generated a viral clearance database that contains product information, unit operation process parameters, and viral clearance data from monoclonal antibody and antibody-related regulatory submissions to FDA. Here we present a broad overview of the database and resulting analyses. We report that the diversity of model viruses tested expands as products transition to late-phase. We also present averages and ranges of viral clearance results by Protein A and ion exchange chromatography steps, low pH chemical inactivation, and virus filtration, focusing on retro- and parvoviruses. For most unit operations, an average log reduction value (LRV, a measure of clearance power) for retrovirus of >4 log(10) were measured. Cases where clearance data fell outside of the anticipated range (i.e., outliers) were rationally explained. Lastly, a historical analysis did not find evidence of any improvement trend in viral clearance over time. The data collectively suggest that many unit operations in general can reliably clear viruses.

  18. Monoclonal antibodies: pharmacokinetics as a basis for new dosage regimens? (United States)

    Azanza, J-R; Sádaba, B; Gómez-Guiu, A


    Complete monoclonal IgG antibodies which are in use in clinical practice share some pharmacological properties resulting in high concentrations in plasma. This fact is reflected in their low volumes of distribution, which can also be correlated with a high molecular weight and water solubility. This feature allows a novel approach to be applied to the dosing schedule for this group of drugs with fixed doses being used instead of the initially developed weight- or body surface-adjusted dosing schedules. In addition, the development of a new formulation containing hyaluronidase allows a subcutaneous route of administration to be used, because hyaluronidase creates a space in the subcutaneous tissue that helps antibody absorption. This method requires higher doses, but has allowed testing the feasibility of administering a fixed dose, with no individual dose adjustments based on weight or body surface. Moreover, loading doses are not needed, because the first dose results, within 3 weeks, in minimum concentrations that are higher than effective concentrations.

  19. Anti-bacterial monoclonal antibodies: back to the future? (United States)

    Oleksiewicz, Martin B; Nagy, Gábor; Nagy, Eszter


    Today's medicine has to deal with the emergence of multi-drug resistant bacteria, and is beginning to be confronted with pan-resistant microbes. This worsening inadequacy of the antibiotics concept, which has ruled infectious medicine in the last six decades creates an increasing unmet medical need that can be addressed by passive immunization. While past experience from the pre-antibiotic era with serum therapy was in many cases encouraging, antibacterial monoclonal antibodies have so far suffered high attrition rates in the clinic, generally from lack of efficacy. Yet, we believe that recent developments in a number of areas such as infectious disease pathogenesis research, translational medicine, mAb engineering, mAb manufacturing and rapid bedside diagnostics are converging to make the medium-term future permissive for antibacterial mAb development. Here, we review antibacterial mAb-based approaches that are or were in clinical development, and may potentially act as paradigms with regards to molecular targets, antibody formats and mode-of-action, pre-clinical validation and selection of most relevant patient populations, in order to increase the likelihood of successful product development in this field.

  20. Defining process design space for monoclonal antibody cell culture. (United States)

    Abu-Absi, Susan Fugett; Yang, LiYing; Thompson, Patrick; Jiang, Canping; Kandula, Sunitha; Schilling, Bernhard; Shukla, Abhinav A


    The concept of design space has been taking root as a foundation of in-process control strategies for biopharmaceutical manufacturing processes. During mapping of the process design space, the multidimensional combination of operational variables is studied to quantify the impact on process performance in terms of productivity and product quality. An efficient methodology to map the design space for a monoclonal antibody cell culture process is described. A failure modes and effects analysis (FMEA) was used as the basis for the process characterization exercise. This was followed by an integrated study of the inoculum stage of the process which includes progressive shake flask and seed bioreactor steps. The operating conditions for the seed bioreactor were studied in an integrated fashion with the production bioreactor using a two stage design of experiments (DOE) methodology to enable optimization of operating conditions. A two level Resolution IV design was followed by a central composite design (CCD). These experiments enabled identification of the edge of failure and classification of the operational parameters as non-key, key or critical. In addition, the models generated from the data provide further insight into balancing productivity of the cell culture process with product quality considerations. Finally, process and product-related impurity clearance was evaluated by studies linking the upstream process with downstream purification. Production bioreactor parameters that directly influence antibody charge variants and glycosylation in CHO systems were identified.

  1. Evaluation of p16, human papillomavirus capsid protein L1 and Ki-67 in cervical intraepithelial lesions: potential utility in diagnosis and prognosis. (United States)

    Alshenawy, Hanan AlSaeid


    Cervical dysplasia, a potentially precancerous lesion, has increased in young women. Detection of cervical dysplasia is important for reducing morbidity and mortality in cervical cancer. This study analyzes the immunohistochemical expression of p16, HPV L1 capsid protein and Ki-67 in cervical intraepithelial lesions, and correlates them with lesion grade to develop a set of markers for diagnosis and detect the prognosis of cervical cancer precursors. Seventy-five specimens were analyzed, including 15 cases of CIN 1, 28 cases of CIN 2, 20 cases of CIN 3, and 12 cervical squamous carcinomas, besides 10 normal cervical tissues. They were stained for p16, HPV L1 and Ki-67. Sensitivity, specificity, predictive values and accuracy were evaluated for each marker. p16 expression increased during progression from CIN 1 to carcinoma. HPV L1 positivity was detected in CIN 2 and decreased gradually as the CIN grade increased but disappeared in carcinoma. Strong Ki-67 expression was observed in high grades CIN and carcinoma. p16, HPV L1 and Ki-67 were sensitive but with variable specificity in detecting CIN lesions. p16, HPV L1 and Ki-67 are useful markers in establishing the risk of high-grade CIN. They complete each other to reach an accurate diagnosis and to detect the prognosis. Copyright © 2014 Elsevier GmbH. All rights reserved.

  2. P53、Ki-67在甲状腺乳头状癌和滤泡癌中的表达及相互关系%The expression and their correlation of p53, Ki-67 in papillary thyroid carcinoma and follicular thyroid carcinoma

    Institute of Scientific and Technical Information of China (English)

    陈洪杰; 孟予城; 孙革


    目的 探讨抑癌基因p53 与细胞核增殖抗原Ki-67 在甲状腺乳头状癌和滤泡癌中的表达与相互关系.方法 应用免疫组织化学法(sp法)检测49例甲状腺癌(包括39例乳头状癌和10例滤泡癌)、30例甲状腺腺瘤组织中p53和Ki-67的表达情况.结果 p53、Ki-67在甲状腺癌中阳性表达率分别为83.6%和69.38%,两者呈正相关(P<0.05).两者与甲状腺腺瘤阳性表达率比较差异有统计学意义(P<0.05).p53、Ki-67在甲状腺癌中的阳性率与性别、年龄、肿瘤大小、结节数量无关(P>0.05).在甲状腺乳头状癌和滤泡癌中p53和Ki-67阳性率差异无统计学意义(P>0.05),但Ki-67增殖指数乳头状癌要高于滤泡癌.结论 p53与Ki-67在甲状腺癌中的表达呈正相关,Ki-67在甲状腺乳头状癌和滤泡癌中的增殖指数有差异.%Objective To investigate the expression of P53, Ki-67 and their relationship in papillary thyroid carcinoma and follicular thyroid carcinoma. Methods lmmunohistochemical method (SP) was used to detect the expression of P53 and Ki-67 in 49 cases of thyroid carcinoma including 39 eases of papillary thyroid carcinoma and 10 cases of follicular thyroid carcinoma, as well as 30 eases of thyroid adenoma. Results The positive expression rates of P53 ,Ki-67 were 83.6% and 69.38% respectively in thyroid carcinoma, which were significantly higher than those in thyroid adenoma. The positive expression rate of P53 was positively correlated with that of Ki-67 in thyroid carcinoma( P <0.05). The expressions of P53, Ki-67 were not correlated with gender,age,tumor size and nodule number( P > 0. 05 ). The positive expression rates of P53, Ki-67 were significantly different between papillary thyroid carcinoma and follicular thyroid carcinoma( P > 0.05 ). The Ki-67 proliferation index in papillary thyroid carcinoma was higher than that of follicular thyroid carcinoma. Conclusion The expression of P53 is positively correlated with that of

  3. Ki-67在T1期乳腺癌中的表达及其与Her-2和ER的关系%Ki-67 expression in T1 primary breast cancer tissue and its relationship with Her-2 and ER

    Institute of Scientific and Technical Information of China (English)



    目的 研究Ki-67在T1期乳腺癌的表达情况,以及其与雌激素受体(ER)、人表皮生长因子受体-2(Her-2)的关系,指导早期乳腺癌的预后判断.方法 选择2008年1月至2011年6月的女性新发T1期乳腺癌组织共60例,免疫组化检测Ki-67的表达情况,并分析其表达与临床病理特征和Her-2、ER表达的关系.结果 60例T1期乳腺癌中Ki-67阳性表达率为55.0%,Ki-67表达与T1期乳腺癌的病理类型、患者绝经与否无关(P>0.05).T1期乳腺癌中,Ki-67的表达在有淋巴结转移组高于无淋巴结转移组(P=0.047,P<0.05),Ki-67在Her-2阳性组表达高于阴性组(P=0.021,P<0.05),Ki-67在ER阳性组表达低于ER阴性组(P=0.037,P<0.05).结论 T1乳腺癌中Ki-67阳性表达者淋巴结转移可能性较大,与Her-2阳性、ER阴性表达正相关,提示预后不良.%Objective To study the expression of Ki-67 in T1 primary breast cancer tissue and its relationship with Her-2 and ER,evaluate the prognosis of early breast cancer.Method s Sixty cases of women T1 primary breast cancer tissues of 60 female cases from January 2008 to June 2011 were selected.The expressions of Ki-67 were detected by immunohistochemieal method and the relationships between Ki-67 and clinical-pathological fcatures,Her-2 and ER were analysed.Results The expression of Ki-67 in T1 primary breast cancer was 55% among the 60 patients.The expression of Ki-67 was not related with the clinical-pathological type of breast cancer and menopause (P>0.05).The expression of Ki-67 in group with lymph node metastasis was higer than in group without lymph node metastasis(P =0.047 ,P <0.05).The expression of Ki-67 in Her-2positive group was higher than Her-2 negative group (P= 0.021,P<0.05),and the expression of Ki-67 in ER positive group was lower than that of ER negative group (P = 0.037,P < 0.05).Conclusions The expression of Ki-67 in T1 primary breast cancer indicates the possibility of lymph node metastasis,and its

  4. Generation and applications of monoclonal antibodies for livestock production. (United States)

    Van Der Lende, T


    Monoclonal antibodies (MCAs) have found widespread applications in livestock production. Although the generation of murine MCAs is at present a routine, the production of homologous MCAs, especially important for in vivo applications, is still hampered by the lack of efficient homologous fusion partners for immortalization of antibody producing lymphocytes of livestock species. At present, MCAs are used in immunodiagnostic tests e.g. to monitor livestock reproduction and quality of livestock products. In the future MCAs will also be used in immunosensors for real-time and on-site applications in the same areas. The commercial application of MCAs for the immunomodulation of (pharmacologically induced) physiological processes underlying important (re)production traits is at present limited to the use of anti-PMSG MCAs in PMSG-induced superovulation. However, many potentially interesting applications are under investigation (e.g. immunopotentiation of growth hormone to enhance growth; immunocytolysis of adipocytes to increase lean meat production; immunoneutralization of GnRH for immunocastration; immunoimitation of hormone activity with anti-idiotype antibodies). Attempts to use specific MCAs for the sexing of embryos have been disappointing, mainly because of the relatively low accuracy. In the future, MCAs against membrane proteins which are specific for X- or Y-chromosome bearing spermatozoa might be used for bulk separation of livestock sperm. In general, it is expected that engineered (homologous) recombinant MCAs will largely contribute to the development of a new generation of rapid immunodiagnostic tests and effective immunomodulation applications. They will further increase the use of MCAs in livestock production.

  5. Kinetic analysis of the multistep aggregation mechanism of monoclonal antibodies. (United States)

    Nicoud, Lucrèce; Arosio, Paolo; Sozo, Margaux; Yates, Andrew; Norrant, Edith; Morbidelli, Massimo


    We investigate by kinetic analysis the aggregation mechanism of two monoclonal antibodies belonging to the IgG1 and IgG2 subclass under thermal stress. For each IgG, we apply a combination of size exclusion chromatography and light scattering techniques to resolve the time evolution of the monomer, dimer, and trimer concentrations, as well as the average molecular weight and the average hydrodynamic radius of the aggregate distribution. By combining the detailed experimental characterization with a theoretical kinetic model based on population balance equations, we extract relevant information on the contribution of the individual elementary steps on the global aggregation process. The analysis shows that the two molecules follow different aggregation pathways under the same operating conditions. In particular, while the monomer depletion of the IgG1 is found to be rate-limited by monomeric conformational changes, bimolecular collision is identified as the rate-limiting step in the IgG2 aggregation process. The measurement of the microscopic rate constants by kinetic analysis allows the quantification of the protein-protein interaction potentials expressed in terms of the Fuchs stability ratio (W). It is found that the antibody solutions exhibit large W values, which are several orders of magnitude larger than the values computed in the frame of the DLVO theory. This indicates that, besides net electrostatic repulsion, additional effects delay the aggregation kinetics of the antibody solutions with respect to diffusion-limited conditions. These effects likely include the limited efficiency of the collision events due to the presence of a limited number of specific aggregation-prone patches on the heterogeneous protein surface, and the contribution of additional repulsive non-DLVO forces to the protein-protein interaction potential, such as hydration forces.

  6. Foco de criptas aberrantes e câncer da junção colorretal: análise da presença de lesões precoces microscópicas na periferia do câncer colorretal e correlação com a expressão da β-catenina e Ki-67 Aberrant crypt foci and cancer of the colorectal junction: the correlation between β-catenin/Ki-67 expression and the occurrence of early microscopic secondary lesions surrounding periphery colorectal cancer

    Directory of Open Access Journals (Sweden)

    Daniel Cury Ogata


    : Among all adenocarcinoma samples, the frequency of positive â-catenin nuclear expression was 81%. The Ki-67 expression demonstrated the same percentage of positivity as did β-catenin. However, the Kappa coefficient showed weak relationship between those two antibodies. Among 20 ACF's analyzed, 13 were located close (1 cm to the tumor. None of the ACF's demonstrated nuclear expression of β-catenin or Ki-67. CONCLUSION: Higher concentrations of ACF's were observed in colonic mucosa at a distance of 1 cm compared to samples at 5 cm from the primary CRC. However, we could not demonstrate positive correlation between colonocytes β-catenin expression and the occurrence of ACF's.

  7. Immunolocation of antisperm monoclonal antibody 6B10 and corresponding antigen

    Institute of Scientific and Technical Information of China (English)

    高绍荣; 胡国俊; 段崇文; 刘辉; 韩之明; 宋祥芬; 陈大元


    An antisperm monoclonal antibody 6B10 was produced by hybridoma technique of the isotype IgG. The monoclonal antibody was purified by means of ammonium sulfate precipitation and protein A-Sepharose Cl-4B affinity chromatography. SDS polyacrylamide gel electrophoresis was used to evaluate the purity of the antibody. Evaluation of the sperm acrosomal status was determined by chlortetracycline (CTC) staining. It was found that monoclonal antibody 6B10 can inhibit the sperm acrosome reaction induced by progesterone. The corresponding antigen recognized by monoclonal antibody 6B10 was located on the plasma membrane of the sperm acrosome by indirect immunofluorescent microscopy and immunoelectronmicroscopy. Sperm protein was extracted by 1% Triton X-100. The molecular weight of the antigen is 50 ku, detected by Western blot. The antigen is a key protein in the sperm acrosome reaction and may be the receptor of progesterone on the sperm acrosome. It may either be developed as a candidate contraceptive vaccine

  8. Immunogenicity assessment of monoclonal antibody products: A simulated case study correlating antibody induction with clinical outcomes. (United States)

    Knezevic, Ivana; Kang, Hye-Na; Thorpe, Robin


    Monoclonal antibodies are large molecules with complex structure and functions. They have a wide application for treatment of a broad range of chronic diseases and represent the largest class of biotherapeutic products. Given that biotherapeutic products may induce unwanted humoral and/or cellular immune responses in recipients, it is essential to investigate the immunogenicity of a product prior to licensure. The immune response is influenced by many factors and data generated in the pre-licensure studies are usually somewhat difficult for regulatory review. The knowledge and expertise required for this requires a thorough understanding of animal and human immunology as well as specific product characteristics, including mechanism of action, antibody assays and assessment of results in a given clinical context. The appropriate interpretation of immunogenicity data is of critical importance for defining the safety profile of a monoclonal antibody. Two case studies described in this paper were prepared to mimic a real situation in which regulators need to evaluate immunogenicity studies conducted by manufacturers of monoclonal antibody products. The specific objective of the case studies was to illustrate assessment of unwanted immunogenicity and the important factors that need to be considered in this context. Regulators and manufacturers who attended the World Health Organization (WHO) implementation workshop on Evaluation of Biotherapeutic Products, held in Seoul, Republic of Korea, in May 2014, participated in the case studies and provided valuable input. This article outlines the main aspects of immunogenicity discussed in these case studies and a summary of the lessons learned at this occasion. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  9. Monoclonal antibodies neutralizing the haemolytic activity of box jellyfish (Chironex fleckeri) tentacle extracts. (United States)

    Collins, S P; Comis, A; Marshall, M; Hartwick, R F; Howden, M E


    1. Three monoclonal antibodies have been produced which neutralize in vitro the haemolytic activity present in tentacle extracts of the box jellyfish (Chironex fleckeri). 2. Two of these monoclonal antibodies bound specifically to a component of relative molecular mass 50,000 in tentacle extract on Western blots. 3. This binding only occurred when the extracts were electrophoresed under non-reducing conditions. 4. The third monoclonal antibody did not display binding to Western blots of tentacle extract under any of our experimental conditions.

  10. Radiolabeled monoclonal antibodies for imaging and therapy: Potential, problems, and prospects: Scientific highlights

    Energy Technology Data Exchange (ETDEWEB)

    Srivastava, S.C.; Buraggi, G.L.


    This meeting focused on areas of research on radiolabeled monoclonal antibodies. Topics covered included the production, purification, and fragmentation of monoclonal antibodies and immunochemistry of hybridomas; the production and the chemistry of radionuclides; the radiohalogenation and radiometal labeling techniques; the in-vivo pharmacokinetics of radiolabeled antibodies; the considerations of immunoreactivity of radiolabeled preparations; the instrumentation and imaging techniques as applied to radioimmunodetection; the radiation dosimetry in diagnostic and therapeutic use of labeled antibodies; the radioimmunoscintigraphy and radioimmunotherapy studies; and perspectives and directions for future research. Tutorial as well as scientific lectures describing the latest research data on the above topics were presented. Three workshop panels were convened on ''Methods for Determining Immunoreactivity of Radiolabeled Monoclonal Antibodies - Problems and Pitfalls,'' Radiobiological and Dosimetric Considerations for Immunotherapy with Labeled Antibodies,'' and ''The Human Anti-Mouse Antibody Response in Patients.''

  11. Ki-67 cell proliferation in familial and in esporadic unilateral retinoblastoma: case report Avaliação da proliferação celular com marcador ki-67 em pacientes com retinoblastoma unilateral esporádico e familiar: relato de caso

    National Research Council Canada - National Science Library

    Maria Teresa Brizzi Chizzotti Bonanomi; Fernando Betty Cresta; Clayton César Tokunaga; Ruth Miyuki Santo; Otacilio de Oliveira Maia; Walter Yukihiko Takahashi


    .... This study is a clinicopathologic observational case report that aims to report on the cell proliferation rates, as measured by the Ki-67 antigen, in two enucleated retinoblastoma eyes. METHODS...

  12. Use of AN Eosinophil Specific Monoclonal Antibody in Assessing Eosinophil Function. (United States)

    Minkoff, Marjorie Sue

    A monoclonal antibody to an eosinophil specific determinant is very important in assessing eosinophil function during helminthic infection. Eosinophils induced by Schistosoma mansoni infection in BALB/c mice were used to induce C57B1/6 immunocytes for production of hybridomas secreting eosinophil monoclonal antibodies. These antibodies were shown to react with an eosinophil surface epitope but not with neutrophils or macrophages as determined by ELISA, immunodiffusion, immunofluorescence, and immunoblot assay. Affinity chromatography with eosinophil chemotactic factor-sepharose consistently selected out a { rm M_ R} 67,000 protein from solubilized eosinophil membrane antigens but not from neutrophil and macrophage antigens. In vitro studies showed that the eosinophil-specific monoclonal antibodies abrogated antibody-dependent eosinophil -mediated killing of S. mansoni schistosomula using mouse, rat or human eosinophils. Neutrophil and macrophage killing activities were unaffected. The monoclonal antibodies effected complement-dependent lysis of mouse and rat eosinophils but not of human eosinophils. ECF-treated eosinophils showed enhanced killing of schistosomula which was blocked by the monoclonal antibody. Murine and human eosinophils preincubated with monoclonal antibody exhibited decreased chemotaxis to ECF at optimal chemotactic concentrations. The monoclonal antibody also blocked eosinophil binding to ECF- sepharose beads. In vivo induction of peripheral blood eosinophilia by injection of S. mansoni eggs was suppressed by injections of monoclonal antibodies 2CD13 and 2QD45 in mouse and rat experimental models. Eosinophilia induced by keyhole limpet hemocyanin- cyclophosphamide treatment was also suppressed by monoclonal antibody in both murine and rat systems. Pulmonary granulomas in mice given egg injection and monoclonal antibody were smaller and contained fewer eosinophils than those granulomas from mice given eggs only. In immuno-biochemical studies, the

  13. Changes in the ER, PgR, HER2, p53 and Ki-67 biological markers between primary and recurrent breast cancer: discordance rates and prognosis

    Directory of Open Access Journals (Sweden)

    Tashima Rumiko


    Full Text Available Abstract Background In breast cancer, ER/PgR, HER2, and Ki-67 are important biological markers for predicting prognosis and making effective treatment decisions. In addition, changes in markers due to relapse are also clinically experienced; however, the frequency and clinical significance are still not fully understood. Thus, changes in markers and their correlations with prognosis were investigated. Patients and Methods Out of the patients with relapse from 1997 to March 2011, there were 97 consecutive patients from whom the lesion was resected and evaluated by immunostaining. The biopsy sites were chest wall, lymph node, ipsilateral breast tumor recurrence, lungs, bones, ovaries and brain. The markers sought were ER, PgR, HER2, p53 and Ki-67. Results The hormone receptor positive rate from the primary tumor to recurrence decreased from 63.9% to 57.7% and from 56.7% to 43.3% for ER and PgR, respectively. Changes in the positive/negative evaluation were seen at the rate of 10.3% and 25.8% for ER and PgR, respectively. The Ki-67 index increased significantly from a mean of 29.1% at primary tumor to 36.3% at relapse. When divided into 2 groups ( Conclusion Estrogen receptor and PgR decreased while Ki-67 increased due to relapse; however, the rate of change was high for PgR and Ki-67. Change in the subtypes was seen in 25%. In addition, PgR at relapse and Ki-67 at primary tumor were significant factors for post-relapse prognosis while PgR becoming negative was a poor prognostic factor. These findings are important for making effective treatment decisions.

  14. p16/Ki-67 co-expression associates high risk human papillomavirus persistence and cervical histopathology: a 3-year cohort study in China. (United States)

    Yu, Lu-Lu; Guo, Hui-Qin; Lei, Xiao-Qin; Qin, Yu; Wu, Ze-Ni; Kang, Le-Ni; Zhang, Xun; Qiao, You-Lin; Chen, Wen


    To evaluate the association of p16/Ki-67 co-expression and persistence of high-risk human papillomavirus (HR-HPV) infection as well as cervical abnormalities. We performed a 3-year cohort study among which 2498 Chinese women aged 25 to 65 years were screened by different HPV tests in 2011. 690 women who were positive at any of the tests and a random sample of 164 women with all negative results received colposcopy, cervical specimens for cobas HPV test (Roche diagnostics) were collected before colposcopy; of this group, 737 cervical specimens were collected to perform cobas, Liquid-based cytology, HPV E6 test (Arbor Vita Corporation) and p16/Ki-67 dual staining (Roche diagnostics) in 2014. Colposcopy and biopsies was performed on women with any abnormal result. Compared to women without HR-HPV persistent infection, women in the HR-HPV persistence group had a higher risk of p16/Ki-67 positive, with an adjusted Odds Ratio(OR) and 95% confidence interval (CI) of 6.29 (4.07-9.72); moreover, adjusted odds ratio for women who had HPV16/18 persistent infection was nearly 4-folder higher than women with other 12 HR-HPV persistent infection (adjusted OR = 17.15, 95% CI: 7.11-41.33 vs adjusted OR = 4.68, 95% CI: 2.89-7.58). Additionally, p16/Ki-67 positivity rate significantly increased with the severity of the cytological and histological abnormalities, and resulted strongly associated with a CIN2+ diagnosis (OR = 16.03, 95% CI: 4.46-57.59). p16/Ki-67 co-expressions associated strongly with HR-HPV persistence, especially with HPV16/18, and the presence of a CIN2+ lesion. Therefore, p16/Ki-67 could be considered as a suitable biomarker for cervical cancer screening, particularly in HPV-based screening programs.

  15. p16 INK4a and Ki67 expression in normal, dysplastic and neoplastic uterine cervical epithelium and human papillomavirus (HPV) infection. (United States)

    Calil, L N; Edelweiss, M I A; Meurer, L; Igansi, C N; Bozzetti, M C


    Cellular cycle proteins like the p16(INK4a) and the Ki67 proliferation nuclear antigen have been used as oncogenicity cellular markers. The E6 and E7 oncoproteins interact with tumor suppressor genes p53 and pRb, culminating with the p16(INK4a) overexpression. The objective of this study was to evaluate the presence of HPV-DNA in 174 cervical biopsies and correlate the different histological grades with the p16(INK4a) and Ki67 immunohistochemical expression (IHC). A cross-sectional study that enrolled a total of 174 women who underwent uterine cervical biopsies between February 2003 and December 2006, in southern Brazil, was performed. Cervical smear samples were analyzed for the presence of HPV-DNA through polymerase chain reaction (PCR), and biopsy samples were examined for p16(INK4A) and Ki67 expression through IHC techniques. The presence of HPV-DNA was observed in 89% of the tested patients, among which 52% were positive for high-risk (HR) viral types [16, 18 and 31]. Regarding p16(INK4a), an expression of 69% was observed, being expressed in 100% of the high-grade squamous lesions (HSIL) and HR-HPV-DNA positives. Ki67 expression was associated with the lesion grade, being more expressive in the most severe lesions (pp16(INK4A) and Ki67 markers coexpression was present in 86% of the samples (pp16(INK4a) and Ki67 expression and which is even stronger among women with HSIL. Copyright © 2014 Elsevier GmbH. All rights reserved.

  16. Evaluation of the p16 and Ki-67 Biomarkers as Predictors of the Recurrence of Premalignant Cervical Cancer Lesions after LEEP Conization. (United States)

    Leite, Paulo Macêdo de Oliveira; Tafuri, Luciene; Costa, Maria Zélia de Oliveira; Lima, Maria Inês de Miranda; Simões, Renata Toscano


    Objective To evaluate the expressions of biomarkers p16 and Ki-67 in low-grade (LG) or high-grade (HG) lesions, and to relate them to risk factors and the recurrence of these lesions. Methods A retrospective case-control study of 86 patients with LG and HG lesions who underwent a loop electrosurgical excision procedure (LEEP) between 1999 and 2004. The control group was composed of 69 women with no recurrence, and the study group, of 17 patients with recurrence. All patients were followed-up over a two-year period after surgery, and screened every six months, including cytology and colposcopy. Biopsy samples collected from LEEP were submitted to immunohistochemical analysis for p16 and Ki-67. The statistical analysis was performed using the Statistical Package for the Social Sciences software (SPSS, IBM-SPSS, Inc., Chicago, IL, US), with a significant p ​​p16 and Ki-67, separately or combined, showed no relation to recurrence on the total analysis. However, evaluating specifically HG lesions, the positive expression (2+ and 3+) of p16/Ki-67 was associated with recurrence (0.010). In addition, p16 isolated was also more expressive in HG lesions (2+ and 3 + , p = 0.018), but it was unrelated to recurrence. Conclusion Proteins p16 and Ki-67, both isolated and combined, are not reliable primary markers for the recurrence of cervical lesions in the majority of LG lesions. However, analyzing only the group with prior diagnosis of HG lesions, the expressions of p16 and of p16/Ki-67 were associated with recurrence, and they may be useful in monitoring these cases. Thieme-Revinter Publicações Ltda Rio de Janeiro, Brazil.

  17. Prognostic and clinicopathological role of high Ki-67 expression in patients with renal cell carcinoma: a systematic review and meta-analysis (United States)

    Xie, Yongpeng; Chen, Luyao; Ma, Xin; Li, Hongzhao; Gu, Liangyou; Gao, Yu; Fan, Yang; Zhang, Yu; Zhang, Xu


    Previous studies have elevated the prognostic value of Ki-67 in renal cell carcinoma (RCC), but the reports are controversial and inconsistent. We conducted a systematic review and meta-analysis to clarify the significance of Ki-67 in RCC prognosis. We systematically searched PubMed, Web of Science, and Embase to identify relevant studies until April 2016. Based on the inclusion and exclusion criteria, 20 studies, including 5,398 patients, were eligible for further analysis. Results showed that high Ki-67 expression in RCC was associated with poor OS (HR = 1.95, 95% CI: 1.44–2.64), CSS (HR = 1.67, 95% CI: 1.47–1.89), and DFS (HR = 2.56, 95% CI: 1.79–3.67). In addition, high Ki-67 expression was significantly associated with TNM stage (III/IV vs. I/II: RR = 2.03, 95% CI: 1.68–2.44), pathological T stage (T3/T4 vs. T1/T2: RR = 1.67, 95% CI: 1.35–2.06), metastasis (yes vs. no: RR = 2.15, 95% CI: 1.77–2.62), and Fuhrman grade (III/IV vs. I/II: RR = 1.77, 95% CI: 1.20–2.60). Our study suggested that Ki-67 was a prognostic marker in RCC. High Ki-67 expression was correlated with poor prognosis and advanced clinicopathological features, and it could serve as a biomarker for disease management. PMID:28287186

  18. Identification of antigen-specific human monoclonal antibodies using high-throughput sequencing of the antibody repertoire. (United States)

    Liu, Ju; Li, Ruihua; Liu, Kun; Li, Liangliang; Zai, Xiaodong; Chi, Xiangyang; Fu, Ling; Xu, Junjie; Chen, Wei


    High-throughput sequencing of the antibody repertoire provides a large number of antibody variable region sequences that can be used to generate human monoclonal antibodies. However, current screening methods for identifying antigen-specific antibodies are inefficient. In the present study, we developed an antibody clone screening strategy based on clone dynamics and relative frequency, and used it to identify antigen-specific human monoclonal antibodies. Enzyme-linked immunosorbent assay showed that at least 52% of putative positive immunoglobulin heavy chains composed antigen-specific antibodies. Combining information on dynamics and relative frequency improved identification of positive clones and elimination of negative clones. and increase the credibility of putative positive clones. Therefore the screening strategy could simplify the subsequent experimental screening and may facilitate the generation of antigen-specific antibodies.

  19. A monoclonal antibody that recognizes an antigenic determinant shared by HLA A2 and B17. (United States)

    McMichael, A J; Parham, P; Rust, N; Brodsky, F


    A hybridoma monoclonal anti-HLA antibody has been produced by the technique of Kohler and Milstein [1]. This antibody recognizes a new specificity common to HLA A2 and B17. It was shown to be a single antibody by isoelectric focusing and absorption experiments.


    NARCIS (Netherlands)



    In this study, the effect of antibody adsorption on physicochemical properties of Streptococcus sobrinus was studied. Bacteria were preincubated with polyclonal antibodies or with OMVU10, a monoclonal antibody (MAb) reactive with S. sobrinus. The zeta potentials and the hydrophobicity as determined

  1. Predictive value of p16/Ki-67 immunocytochemistry for triage of women with abnormal Papanicolaou test in cervical cancer screening: a systematic review and meta-analysis. (United States)

    Chen, Cheng-Chieh; Huang, Lee-Wen; Bai, Chyi-Huey; Lee, Chin-Cheng


    The Papanicolaou (Pap) test is one screening strategy used to prevent cervical cancer in developed countries. The p16/Ki-67 immunocytochemistry is a triage test performed on Pap smears in women with atypical squamous cells of undetermined significance (ASCUS) or low grade squamous intraepithelial lesion. Our objective was to review studies investigating the diagnostic performance of p16/Ki-67 dual stain for triage of women with abnormal Pap tests. We conducted a systematic review and meta-analysis of diagnostic test accuracy studies. We followed the protocol of systematic review of diagnostic accuracy studies. We searched PubMed, The Cochrane Library, BioMed Central, and for relevant studies. We included research that assessed the accuracy of p16/Ki-67 dual stain and high risk human papillomavirus testing for triage of abnormal Pap smears. Review articles and studies that provided insufficient data to construct 2.2 tables were excluded. Data synthesis was conducted using a random-effects model. Sensitivity and specificity. In seven studies encompassing 2628 patients, the pooled sensitivity and specificity of p16/Ki-67 for triage of abnormal Pap smear results were 0.91 (95% CI, 0.89 to 0.93) and 0.64 (95% CI, 0.62 to 0.66), respectively. No study used a case-control design. A subgroup analysis involving liquid-based cytology showed a sensitivity of 0.91 (95%CI, 0.89 to 0.93) and specificity of 0.64 (95%CI, 0.61 to 0.66). Our meta-analysis of p16/Ki-67 dual stain studies showed that the test achieved high sensitivity and moderate specificity for p16/Ki-67 immunocytochemistry for high-grade squamous intraepi.thelial lesion and cervical cancer. We suggest that p16/Ki-67 dual stain might be a reliable ancillary method identifying high-grade squamous intraepithelial lesions in women with abnormal Pap tests. No study in the meta-analysis examined the accuracy of the p16/Ki-67 dual stain for inter.pretation of glandular neoplasms.

  2. Clinical value of fully automated p16/Ki-67 dual staining in the triage of HPV-positive women in the Norwegian Cervical Cancer Screening Program. (United States)

    Ovestad, Irene T; Dalen, Ingvild; Hansen, Elisabeth; Loge, Janne L D; Dybdahl, Britt Mona; Dirdal, Marius B; Moltu, Pia; Berland, Jannicke M


    More accurate biomarkers in cervical cytology screening could reduce the number of women unnecessarily referred for biopsy. This study investigated the ability of p16/Ki-67 dual staining to predict high-grade cervical intraepithelial neoplasia (CIN) in human papillomavirus (HPV)-positive women from the Norwegian Cervical Cancer Screening Program. Automated p16/Ki-67 dual staining was performed on liquid-based cytology samples from 266 women who were HPV-positive at their secondary screening. At a mean of 184 days after p16/Ki-67 staining, 201 women had a valid staining result and a conclusive follow-up diagnosis (histological diagnosis or HPV-negative diagnosis with normal cytology findings). The sensitivity and specificity for predicting the follow-up diagnosis were compared for cytology, p16/Ki-67 dual staining, and their combination. Sixty-seven percent of the study sample was p16/Ki-67-positive. The sensitivity of p16/Ki-67 staining for predicting CIN-2/3 was statistically significantly higher than the sensitivity of cytology (0.88 vs 0.79; P = .008), but this was not true for the prediction of CIN-3 (0.94 vs 0.88; P = .23). The specificity of cytology for predicting CIN-3 was significantly higher than the specificity of p16/Ki-67 staining (0.35 vs 0.28; P = .002), but this was not true for CIN-2/3 (0.35 vs 0.31; P = .063). For predicting CIN-2/3 and CIN-3, combination testing gave potentially better sensitivity (0.95 and 0.96, respectively) and better specificity (0.49 and 0.50, respectively). In a population of HPV-positive women, p16/Ki-67 dual staining was more sensitive but less specific than cytology for predicting high-grade CIN. The advantage of using both tests in different combinations is the potential for increasing the specificity or sensitivity in comparison with both methods performed individually. Cancer Cytopathol 2017;125:283-291. © 2016 American Cancer Society. © 2016 American Cancer Society.

  3. ERCC1 and Ki67 in Small Cell Lung Carcinoma and Other Neuroendocrine Tumors of the Lung Distribution and Impact on Survival

    DEFF Research Database (Denmark)

    Skov, Birgit Guldhammer; Holm, B.; Erreboe, A.


    Background: Excision repair cross-complementation group 1 (ERCC1) is a key component of the platinum-DNA repair mechanism. Ki67 is associated with the clinical course of several malignancies. The associations of ERCC1 and Ki67, clinical features and survival in small cell lung carcinoma (SCLC......), typical carcinoid (TC), atypical carcinoid (AC), and large cell neuroendocrine carcinoma (LCNEC) were determined. Materials and Methods: We included a consecutive series of 186 patients with SCLC treated with platinum-based chemotherapy and surgically treated patients with TC (n = 48), AC (n = 15...

  4. Significance of cyclin E, pl6ink4a and ki67 Overexpression in Cervical Exfoliated-cell Specimens for Primary Screening of HPV-related Cervical Carcinoma

    Institute of Scientific and Technical Information of China (English)

    赵富玺; 郭俊成; 崔克; 熊思东


    The aim of this study is to investigate cyclin E, pl6inkda and ki67 as possible diagnostic biomarkers for cervical preneoplasia using cervical exfoliated-cell specimens, and evaluate the significance for screening patients at high risk of developing cervical carcinoma. The expression of cyclin E, pl6inkda and ki67 was examinated in 78 cervical exfoliated epithelial specimens diagnosed as atypical squamous cells of undetermined significance (ASCUS) (12 cases), cervical intraepithelial neoplasia (CIN) of type 1 (17 cases), CIN2_3 (38 cases) and invasive carcinoma (11 cases) using immunohistochemical analysis, and simultaneously, the DNA status of human papillomavims (HPY) type 16/18 was detected by polymerase chain reaction (PCR) using type specific primers, cyclin E, pl6inkda and ki67 were all overexpressed in CINs and invasive carcinoma, compared with little expression in ASCUS ( P <0.005). Overexpression of cyclin E was observed in CIN1(94.1%, X2 = 21.16, P < 0.01), and p16inkda and ki67 were overexpressed in invasive carcinoma (100% and 90.9% respectively). The degree of pl6inkda and ki67 expression correlated well with that of epithelial lesions ( P < 0.005). HPV16/18 infection was assessed in C1Ns and invasive carcinoma samples, and revealed a significant relationship with the degree of cervical epithelial lession. The expression level of pl6inkda and ki67 seemed more closely associated with HPVI6 infection than that of cyclin E (rs= 1.0 vs rs=0.4). Only 1 case in CINI and d cases in CIN2-3 of HPV18 positive samples were detected. Therefore no statistical significance was found by statistical analysis. Overexpression of cyclin E, pl6inkda and ki67 in CINs and invasive carcinoma cells demonstrates the potential use of cyclin E, pl6inkda and ki67 as diagnostic biomarkers for HPV-related cervical neoplastic lesions. In addition, this technique can be used for screening patients at high risk of developing cervical carcinoma.

  5. Good performance of p16/ki-67 dual-stained cytology for surveillance of women treated for high-grade CIN. (United States)

    Polman, Nicole J; Uijterwaal, Margot H; Witte, Birgit I; Berkhof, Johannes; van Kemenade, Folkert J; Spruijt, Johan W M; van Baal, W Marchien; Graziosi, Peppino G C M; van Dijken, Dorenda K E; Verheijen, René H M; Helmerhorst, Theo J M; Steenbergen, Renske D M; Heideman, Daniëlle A M; Ridder, Ruediger; Snijders, Peter J F; Meijer, Chris J L M


    Women treated for high-grade cervical intraepithelial neoplasia (CIN) are at risk of recurrent CIN Grade 2 or worse (rCIN2+). Currently, posttreatment monitoring is performed using cytology or cytology/high-risk (hr)HPV cotesting. This study aimed to evaluate the performance of p16/Ki-67 dual-stained cytology (p16/Ki-67) for posttreatment monitoring. Three hundred and twenty-three women treated for high-grade CIN in the SIMONATH study underwent close surveillance by cytology, hrHPV and DNA methylation marker testing up to 12 months posttreatment. Histological endpoints were ascertained by colposcopy with biopsy at 6 and/or 12 months. p16/Ki-67 dual-staining was performed on residual liquid-based cytology samples obtained at, or shortly before biopsy collection. Clinical performance estimates of cytology, hrHPV, p16/Ki-67 testing and combinations thereof for the detection of rCIN2+ were determined and compared to each other. Sensitivity of p16/Ki-67 for rCIN2+ (69.2%) was nonsignificantly lower than that of cytology (82.1%; ratio 0.84, 95% CI: 0.71-1.01), but significantly lower than that of hrHPV testing (84.6%; ratio 0.82, 95% CI: 0.68-0.99). Specificity of p16/Ki-67 for rCIN2+ (90.4%) was significantly higher compared to both cytology (70.8%; ratio 1.28, 95% CI: 1.19-1.37) and hrHPV testing (76.2%; ratio 1.19, 95% CI: 1.12-1.26). Overall, hrHPV testing showed very high sensitivity, along with a good specificity. When considering cotesting, combined p16/Ki-67/hrHPV testing showed rCIN2+ sensitivity comparable to cytology/hrHPV cotesting (87.2% vs. 89.7%; ratio 0.97, 95% CI: 0.92-1.03), but with significantly increased specificity (74.2% vs. 58.1%; ratio 1.28, 95% CI: 1.19-1.38). Thus, when considered in combination with hrHPV, p16/Ki-67 might be an attractive approach for surveillance of women treated for high-grade CIN.

  6. Evaluating TIMP-1, Ki67, and HER2 as markers for non-sentinel node metastases in breast cancer patients with micrometastases to the sentinel node

    DEFF Research Database (Denmark)

    Tvedskov, Tove Filtenborg; Bartels, Annette; Jensen, Maj-Britt;


    between 2001 and 2007 at the Department of Breast Surgery, Herlev Hospital. The study was designed as a matched case-control study with 25 cases with micrometastases to SN and, in addition, metastatic spread to NSN and 50 matched controls with micrometastases to SN, but without NSN metastases. Patient...... and tumor characteristics were retrieved from the Danish Breast Cancer Cooperative Group database. Immunohistochemical analyses of TIMP-1 and Ki67 and measurements of HER2 on formalin-fixed paraffin-embedded tumor tissue were performed. No significant differences in the immunoreactivity of TIMP-1 and Ki67...

  7. Bcl-2和Ki-67在眼附属器MALT淋巴瘤中的表达及其对预后的影响

    Institute of Scientific and Technical Information of China (English)

    张慧; 钱江; 陈荣家


    目的:探讨Bcl-2和Ki-67在眼附属器MALT淋巴瘤中的表达情况及其对患者预后的影响.方法:收集2001-12/2005-02在我院收治的眼附属器MALT淋巴瘤17例,采用免疫组织化学的方法检测Bcl-2和Ki-67蛋白的表达情况.对患者进行随访,分析Bcl-2和Ki-67蛋白的表达情况与临床病理特征及生存状况的关系.结果:17例眼附属器MALT淋巴瘤中有12例(70.6%)表达Bcl-2蛋白阳性,有10例(58.8%)呈Ki-67蛋白高表达.Bcl-2蛋白表达与Ann Arbor分期明显相关(P<0.05),Ki-67蛋白表达与年龄、Ann Arbor分期明显相关(P<0.05).Bcl-2阳性组、阴性组平均生存期分别为29.8月和62.4月,Ki-67高表达组、低表达组平均生存期分别为27.2月和58.9月.生存分析显示Bcl-2阴性组、Ki-67低表达组生存期较长,Kaplan Metier生存曲线明显不同(P<0.05).结论:Bcl-2和Ki-67是评价眼附属器MALT淋巴瘤生物学行为的参考指标,并对肿瘤的预后有较好的指导意义.

  8. Use of radiolabeled monoclonal antibodies for diagnostic imaging

    Energy Technology Data Exchange (ETDEWEB)

    Endo, Keigo (Kyoto Univ. (Japan). Faculty of Medicine)


    Monoclonal antibodies (MoAbs) are expected to carry radionuclides selectively to target tissues and to offer antigen-specific diagnosis. Indium (In)-111 has many favorable nuclear properties and is efficiently labeled with MoAbs using DAPA as a bifunctional chelating agent. In-111 labeled MoAbs are clinically employed for the diagnosis of malignant melanoma, colorectal cancer and acute myocardial infarction in Japan. Although non-specific deposit of In-111 was seen in liver and bone-marrow, scintigraphy using In-111 labeled MoAbs was encouraging, since it detected about 80% of tumors, tumors missed by conventional diagnostic methods such as CT, and tumors in patients with normal serum CEA values, and acute myocarditis as well as acute myocardial infarction was positive with In-111 labeled Fab fraction of anti-myosin Ab. Acute or subacute toxicity was not observed. Human anti-murine antibody (HAMA) was detected in 53 of 64 (82.8%) patients who were intravenously administered with 20 to 42 mg of anti-melanoma or anti-CEA MoAbs (whole IgG). In contrast, only 5 of 406 (1.2%) patients had detectable levels of HAMA in their serum after receiving 0.5 mg of Fab fraction of MoAb. Recently mouse-human chimeric Ab has been produced by recombinant DNA techniques, which localized well in xenografted tumors and seems to be promising for clinical use. Investigations are under way to increase the tumor to non-tumor ratio by modifying chelating agents for coupling MoAbs with radionuclides. (author).

  9. Characterization of monoclonal antibodies that strongly inhibit Electrophorus electricus acetylcholinesterase. (United States)

    Remy, M H; Frobert, Y; Grassi, J


    In this study, we describe three different monoclonal antibodies (mAbs Elec-403, Elec-408, and Elec-410) directed against Electrophorus electricus acetylcholinesterase (AChE) which were selected as inhibitors for this enzyme. Two of these antibodies (Elec-403 and Elec-410), recognized overlapping but different epitopes, competed with snake venom toxin fasciculin for binding to the enzyme, and thus apparently recognized the peripheral site of AChE. In addition, the binding of Elec-403 was antagonized by 1,5-bis(4-allyldimethylammoniumphenyl)pentan-3-one dibromide (BW284C51) and propidium, indicating that the corresponding epitope encompassed the anionic site involved in the binding of these low-molecular-mass inhibitors. The third mAb (Elec-408), was clearly bound to another site on the AChE molecule, and its inhibitory effect was cumulative with those of Elec-403, Elec-410, and fasciculin. All mAbs bound AChE with high affinity and were as strong inhibitors with an apparent Ki values less than 0.1 nM. Elec-403 was particularly efficient with an inhibitory activity similar to that of fasciculin. Inhibition was observed with both charged (acetylthiocholine) and neutral substrates (o-nitrophenyl acetate) and had the characteristics of a non-competitive process. Elec-403 and Elec-410 probably exert their effect by triggering allosteric transitions from the peripheral site to the active site. The epitope recognized by mAb Elec-408 has not been localized, but it may correspond to a new regulatory site on AChE.

  10. Characterization of monoclonal antibodies to avian Escherichia coli Iss. (United States)

    Lynne, Aaron M; Foley, Steven L; Nolan, Lisa K


    Colibacillosis accounts for annual multimillion dollar losses in the poultry industry, and control of this disease is hampered by limited understanding of the virulence mechanisms used by avian pathogenic Escherichia coli (APEC). Previous work in our laboratory has found that the presence of the increased serum survival gene (iss) is strongly associated with APEC but not commensal E. coli, making iss and the protein it encodes (Iss) candidate targets of colibacillosis-control procedures. Previously, we produced monoclonal antibodies (MAbs) against Iss to be used as a reagent in studies of APEC virulence and colibacillosis pathogenesis. Unfortunately, the utility of these MAbs was limited because these MAbs exhibited nonspecific binding. It was thought that the lack of specificity might be related to the fact that these MAbs were of the immunoglobulin M (IgM) isotype. In the present study, new MAbs were produced using a different immunization strategy in an effort to generate MAbs of a different isotype. Also, because Iss bears strong similarity to Bor, a lambda-derived protein that occurs commonly among E. coli, MAbs were assessed for their ability to distinguish Iss and Bor. For these studies, the bor gene from an APEC isolate was cloned into an expression vector. The fusion protein expressed from this construct was used to assess the potential of the anti-Iss MAbs produced in the past and present studies to distinguish Bor and Iss. The MAbs produced in this study were of the IgG1 isotype, which appeared to bind more specifically to Iss than previously generated antibodies in certain immunologic procedures. These results suggested that the MAbs generated in this study might prove superior to the previous MAbs as a reagent for study of APEC. However, both MAbs recognized recombinant Iss and Bor, suggesting that any results obtained using anti-Iss MAbs would need to be interpreted with this cross-reactivity in mind.

  11. Human Monoclonal Antibodies Broadly Neutralizing against Influenza B Virus (United States)

    Yasugi, Mayo; Kubota-Koketsu, Ritsuko; Yamashita, Akifumi; Kawashita, Norihito; Du, Anariwa; Sasaki, Tadahiro; Nishimura, Mitsuhiro; Misaki, Ryo; Kuhara, Motoki; Boonsathorn, Naphatsawan; Fujiyama, Kazuhito; Okuno, Yoshinobu; Nakaya, Takaaki; Ikuta, Kazuyoshi


    Influenza virus has the ability to evade host immune surveillance through rapid viral genetic drift and reassortment; therefore, it remains a continuous public health threat. The development of vaccines producing broadly reactive antibodies, as well as therapeutic strategies using human neutralizing monoclonal antibodies (HuMAbs) with global reactivity, has been gathering great interest recently. Here, three hybridoma clones producing HuMAbs against influenza B virus, designated 5A7, 3A2 and 10C4, were prepared using peripheral lymphocytes from vaccinated volunteers, and were investigated for broad cross-reactive neutralizing activity. Of these HuMAbs, 3A2 and 10C4, which recognize the readily mutable 190-helix region near the receptor binding site in the hemagglutinin (HA) protein, react only with the Yamagata lineage of influenza B virus. By contrast, HuMAb 5A7 broadly neutralizes influenza B strains that were isolated from 1985 to 2006, belonging to both Yamagata and Victoria lineages. Epitope mapping revealed that 5A7 recognizes 316G, 318C and 321W near the C terminal of HA1, a highly conserved region in influenza B virus. Indeed, no mutations in the amino acid residues of the epitope region were induced, even after the virus was passaged ten times in the presence of HuMAb 5A7. Moreover, 5A7 showed significant therapeutic efficacy in mice, even when it was administered 72 hours post-infection. These results indicate that 5A7 is a promising candidate for developing therapeutics, and provide insight for the development of a universal vaccine against influenza B virus. PMID:23408886

  12. Characterization of monoclonal antibodies against waterfowl parvoviruses VP3 protein

    Directory of Open Access Journals (Sweden)

    Yin Xiuchen


    Full Text Available Abstract Background The VP3 protein of goose parvovirus (GPV or Muscovy duck parvovirus (MDPV, a major structural protein, can induce neutralizing antibodies in geese and ducks, but monoclonal antibodies (MAbs against VP3 protein has never been characterized. Results Three hybridoma cell lines secreting anti-GPV VP3 MAbs were obtained and designated 4A8, 4E2, and 2D5. Immunoglobulin subclass tests differentiated them as IgG2b (4A8 and 4E2 and IgG2a (2D5. Dot blotting assays showed that three MAbs reacted with His-VP3 protein in a conformation-independent manner. A competitive binding assay indicated that the MAbs delineated two epitopes, A and B of VP3. Immunofluorescence assay showed that MAbs 4A8, 4E2, and 2D5 could specifically bind to goose embryo fibroblast cells (GEF or duck fibroblast cells (DEF infected with GPV and MDPV. Dot blotting also showed that the MAbs recognized both nature GPV and MDPV antigen. Western blotting confirmed that the MAbs recognized VP3 proteins derived from purified GPV and MDPV particles. The MAbs 4A8 and 2D5 had universal reactivity to heterologous GPV and MDPV tested in an antigen-capture enzyme-linked immunosorbent assay. Conclusions Preparation and characterization of these the MAbs suggests that they may be useful for the development of a MAb-capture ELISA for rapid detection of both GPV and MDPV. Virus isolation and PCR are reliable for detecting GPV and MDPV infection, but these procedures are laborious, time-consuming, and requiring instruments. These diagnosis problems highlight the ongoing demand for rapid, reproducible, and automatic methods for the sensitive detection of both GPV and MDPV infection.

  13. [ICO-166 monoclonal antibodies against the CD45RA antigen]. (United States)

    Frolova, E A; Baryshnikov, A Iu; Novikov, V V; Syrkin, A B


    Monoclonal antibodies (MCA) ICO-166 against CD45RA antigen were generated and characterized. In the indirect IFA, MCA ICO-166 reacted with 54.1 +/- 1.9% lymphocytes of human peripheral blood and 15.2 +/- 2.3% monocytes but not with granulocytes or thrombocytes. The method of double labelling of cells demonstrated that MCA ICO-166 detected all B-lymphocytes, all NK-cells and 31% of mature T-lymphocytes but only 55% of CD8 suppressor cells and only 21% of CDA helper cells carried this antigen on the surface. Experiments were carried out to block binding of FITC-labeled MCA ALB11 against CD45RA antigen with human lymphocytes by pretreatment of cells with different concentrations of MCA ICO-166. Treatment of cells with MCA ALB11 blocked binding of MCA ALB11-FITC by 85% on the average. MCA ICO-166 blocked binding of MCA ALB11-FITC by 66% on the average. When different dilutions of MCA ICO-166 were used, the dose-dependent effect of blocking of MCA ALB11-FITC binding was observed. MCA ICO-166 immunoprecipitated a protein band of molecular weight 220 kDa from lysates of mononuclear cells of the human peripheral blood.

  14. Role of cosolutes in the aggregation kinetics of monoclonal antibodies. (United States)

    Nicoud, Lucrèce; Sozo, Margaux; Arosio, Paolo; Yates, Andrew; Norrant, Edith; Morbidelli, Massimo


    We propose a general strategy based on kinetic analysis to investigate how cosolutes affect the aggregation behavior of therapeutic proteins. We apply this approach to study the impact of NaCl and sorbitol on the aggregation kinetics of two monoclonal antibodies, an IgG1 and an IgG2. By using a combination of size exclusion chromatography and light scattering techniques, we study the impact of the cosolutes on the monomer depletion, as well as on the formation of dimers, trimers, and larger aggregates. We analyze these macroscopic effects in the frame of a kinetic model based on Smoluchowski's population balance equations modified to account for nucleation events. By comparing experimental data with model simulations, we discriminate the effect of cosolutes on the elementary steps which contribute to the global aggregation process. In the case of the IgG1, it is found that NaCl accelerates the kinetics of aggregation by promoting specifically aggregation events, while sorbitol delays the kinetics of aggregation by specifically inhibiting protein unfolding. In the case of the IgG2, whose monomer depletion kinetics is limited by dimer formation, NaCl and sorbitol are found respectively to accelerate and inhibit conformational changes and aggregation events to the same extent.

  15. Kinetics of Monoclonal Antibody Aggregation from Dilute toward Concentrated Conditions. (United States)

    Nicoud, Lucrèce; Jagielski, Jakub; Pfister, David; Lazzari, Stefano; Massant, Jan; Lattuada, Marco; Morbidelli, Massimo


    Gaining understanding on the aggregation behavior of proteins under concentrated conditions is of both fundamental and industrial relevance. Here, we study the aggregation kinetics of a model monoclonal antibody (mAb) under thermal stress over a wide range of protein concentrations in various buffer solutions. We follow experimentally the monomer depletion and the aggregate growth by size exclusion chromatography with inline light scattering. We describe the experimental results in the frame of a kinetic model based on population balance equations, which allows one to discriminate the contributions of the conformational and of the colloidal stabilities to the global aggregation rate. Finally, we propose an expression for the aggregation rate constant, which accounts for solution viscosity, protein-protein interactions, as well as aggregate compactness. All these effects can be quantified by light scattering techniques. It is found that the model describes well the experimental data under dilute conditions. Under concentrated conditions, good model predictions are obtained when the solution pH is far below the isoelectric point (pI) of the mAb. However, peculiar effects arise when the solution pH is increased toward the mAb pI, and possible explanations are discussed.

  16. DNA immunization as a technology platform for monoclonal antibody induction. (United States)

    Liu, Shuying; Wang, Shixia; Lu, Shan


    To combat the threat of many emerging infectious diseases, DNA immunization offers a unique and powerful approach to the production of high-quality monoclonal antibodies (mAbs) against various pathogens. Compared with traditional protein-based immunization approaches, DNA immunization is efficient for testing novel immunogen designs, does not require the production or purification of proteins from a pathogen or the use of recombinant protein technology and is effective at generating mAbs against conformation-sensitive targets. Although significant progress in the use of DNA immunization to generate mAbs has been made over the last two decades, the literature does not contain an updated summary of this experience. The current review provides a comprehensive analysis of the literature, including our own work, describing the use of DNA immunization to produce highly functional mAbs, in particular, those against emerging infectious diseases. Critical factors such as immunogen design, delivery approach, immunization schedule, use of immune modulators and the role of final boost immunization are discussed in detail.

  17. Characterization of Endotrypanum Parasites Using Specific Monoclonal Antibodies

    Directory of Open Access Journals (Sweden)

    Ramos Franco Antonia Maria


    Full Text Available A large number of Endotrypanum stocks (representing an heterogeneous population of strains have been screened against a panel of monoclonal antibodies (MAbs derived for selected species of Endotrypanum or Leishmania, to see whether this approach could be used to group/differentiate further among these parasites. Using different immunological assay systems, MAbs considered specific for the genus Endotrypanum (E-24, CXXX-3G5-F12 or strain M6159 of E. schaudinni (E-2, CXIV-3C7-F5 reacted variably according to the test used but in the ELISA or immunofluorescence assay both reacted with all the strains tested. Analyses using these MAbs showed antigenic diversity occurring among the Endotrypanum strains, but no qualitative or quantitative reactivity pattern could be consistently related to parasite origin (i.e., host species involved or geographic area of isolation. Western blot analyses of the parasites showed that these MAbs recognized multiple components. Differences existed either in the epitope density or molecular forms associated with the antigenic determinants and therefore allowed the assignment of the strains to specific antigenic groups. Using immunofluorescence or ELISA assay, clone E-24 produced reaction with L. equatorensis (which is a parasite of sloth and rodent, but not with other trypanosomatids examined. Interestingly, the latter parasite and the Endotrypanum strains cross-reacted with a number of MAbs that were produced against members of the L. major-L. tropica complex

  18. Development and Evaluation of Monoclonal Antibodies for Paxilline

    Directory of Open Access Journals (Sweden)

    Chris M. Maragos


    Full Text Available Paxilline (PAX is a tremorgenic mycotoxin that has been found in perennial ryegrass infected with Acremonium lolii. To facilitate screening for this toxin, four murine monoclonal antibodies (mAbs were developed. In competitive indirect enzyme-linked immunosorbent assays (CI-ELISAs the concentrations of PAX required to inhibit signal development by 50% (IC50s ranged from 1.2 to 2.5 ng/mL. One mAb (2-9 was applied to the detection of PAX in maize silage. The assay was sensitive to the effects of solvents, with 5% acetonitrile or 20% methanol causing a two-fold or greater increase in IC50. For analysis of silage samples, extracts were cleaned up by adsorbing potential matrix interferences onto a solid phase extraction column. The non-retained extract was then diluted with buffer to reduce solvent content prior to assay. Using this method, the limit of detection for PAX in dried silage was 15 µg/kg and the limit of quantification was 90 µg/kg. Recovery from samples spiked over the range of 100 to 1000 µg/kg averaged 106% ± 18%. The assay was applied to 86 maize silage samples, with many having detectable, but none having quantifiable, levels of PAX. The results suggest the CI-ELISA can be applied as a sensitive technique for the screening of PAX in maize silage.

  19. Monoclonal antibody probe for assessing beer foam stabilizing proteins. (United States)

    Onishi, A; Proudlove, M O; Dickie, K; Mills, E N; Kauffman, J A; Morgan, M R


    A monoclonal antibody (Mab; IFRN 1625) has been produced, which is specific for the most hydrophobic polypeptides responsible for foam stabilization. The binding characteristics of the Mab suggest that it is the conformation of certain hydrophobic polypeptides which is important for foam stabilization. An enzyme-linked immunosorbent assay (ELISA) for assessing the foam-positive form of the foam-stabilizing polypeptides in beer was developed using IFRN 1625. A good correlation was obtained between ELISA determination of foam-stabilizing polypeptides and an empirical means of determining foaming, that is, the Rudin head retention values, for a collection of beers of various foam qualities. Application of the ELISA to different stages of the brewing process showed that the amounts of foam-positive polypeptides increased during barley germination. During the brewing process the proportion of foam-positive polypeptides present after fermentation increased slightly, although a large amount was lost along with other beer proteins during subsequent steps, such as filtering. The present study demonstrates that the amounts of beer polypeptide present in a foam-positive form have a direct relationship with the foaming potential of beer, that their levels are altered by processing, and that there is potential for greater quality control.

  20. Monoclonal antibodies against NS1 protein of Goose parvovirus. (United States)

    Qiu, Zheng; Tian, Wei; Yu, Tianfei; Li, Li; Ma, Bo; Wang, Junwei


    In the present study, monoclonal antibodies (MAbs) against NS1 protein of Goose parvovirus (GPV) were generated. The secreted MAbs were obtained by fusing mouse myeloma cells and spleen cells of BALB/c mice, which were immunized with the plasmid pcDNA3.1-GPV-NS1 and recombinant protein of GPV-NS1. With indirect ELISA, six hybridoma cell lines against GPV-NS1 were screened. The subtypes of the two MAbs were IgG2a; the others were IgM. The light chain was κ. Western blot analysis showed that six MAbs reacted with recombinant protein GPV-NS1. GPV-NS1 was dissected into 15 overlapping epitopes, which were used to react with MAbs in Western blot. Results showed that six MAbs recognized NS1 protein linear B-cell epitopes located at the C-terminus 453-514 aa, 485-542 aa, and 533-598 aa.

  1. Immunomodulatory Monoclonal Antibodies in Combined Immunotherapy Trials for Cutaneous Melanoma

    Directory of Open Access Journals (Sweden)

    Mariana Aris


    Full Text Available In the last few years, there has been a twist in cancer treatment toward immunotherapy thanks to the impressive results seen in advanced patients from several tumor pathologies. Cutaneous melanoma is a highly mutated and immunogenic tumor that has been a test field for the development of immunotherapy. However, there is still a way on the road to achieving complete and long-lasting responses in most patients. It is desirable that immunotherapeutic strategies induce diverse immune reactivity specific to tumor antigens, including the so-called neoantigens, as well as the blockade of immunosuppressive mechanisms. In this review, we will go through the role of promising monoclonal antibodies in cancer immunotherapy with immunomodulatory function, especially blocking of the inhibitory immune checkpoints CTLA-4 and PD-1, in combination with different immunotherapeutic strategies such as vaccines. We will discuss the rational basis for these combinatorial approaches as well as different schemes currently under study for cutaneous melanoma in the clinical trials arena. In this way, the combination of “push and release” immunomodulatory therapies can contribute to achieving a more robust and durable antitumor immune response in patients.

  2. An update on newer monoclonal antibodies in lymphoma therapy

    Directory of Open Access Journals (Sweden)

    Subhashini Archana Kadavakolan


    Full Text Available In 2014, an estimated 9.4% of all new cancers in the US were accounted to hematological cancers. Most of these cancers have a B-cell origin and on the cell surface express antigen CD20-known to restrict B-cells. Considering the intrinsic immune status of the patients receiving chemotherapy, monoclonal antibodies (mAbs are designed to provide active or passive immunotherapy. Clinical success of rituximab-anti-CD20 mAb in the treatment of lymphoma has led to the development of newer generations of mAb to increase the anti-tumor activity. Hence, recent advances in lymphoma therapy are being built on the conventional prototype of anti-CD20 mAb-rituximab. Our review is an update on the advances in lymphoma therapy using mAb against CD20 including the second generation-ofatumumab, veltuzumab, ocrelizumab, and the third-generation mAbs-ocaratuzumab and obinutuzumab.

  3. Establishment of a novel monoclonal antibody against LGR5. (United States)

    Sasaki, Yuka; Kosaka, Hiromichi; Usami, Katsuaki; Toki, Hiroe; Kawai, Hironori; Shiraishi, Norihiko; Ota, Toshio; Nakamura, Kazuyasu; Furuya, Akiko; Satoh, Mitsuo; Hasegawa, Kazumasa; Masuda, Kazuhiro


    LGR5 is an orphan G-protein-coupled receptor (GPCR) that is expressed on the cell surface membrane. LGR5 is reported to be overexpressed in colon, liver, and ovary tumor compared to normal tissue. However, a specific ligand for LGR5 has not yet been determined, and the function is still not clear. An LGR5-specific monoclonal antibody (mAb) is needed as a tool for detection and analysis of LGR5 biological function and cancer therapy. To date, no mAb against LGR5 that retains high affinity and specificity has been reported. Here, we report successful establishment and characterization of a mAb (KM4056) that specifically recognizes the extracellular N-terminal domain of human LGR5, but not LGR4 or LGR6. This mAb has potent complement-dependent cytotoxicity (CDC) activity in vitro and shows strong anti-tumor activity in vivo against xenograft model by transplanting LGR5 expressing CHO transfectants into SCID mice. Thus, KM4056 can be a useful tool for detection of LGR5 positive cells and analysis of LGR5 biological function.

  4. Production and Characterization of Monoclonal Antibody Against Recombinant Human Erythropoietin

    Institute of Scientific and Technical Information of China (English)



    Objective To produce specific monoclonal antibody(mAb)against recombinant human erythropoietin(rHuEPO)for development of higmy efficient methods for erythropoietin detection in biological fluids.Methods rHuEPO was covalently coupled with bovine serum albumin(BSA)and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology.The obtained F3-mAb was characterized by enzyme-linked immunosorbent assay (ELISA),SDS-PAGE and Western blot.Results The isotype of F3-mAb Was found to be IgM with an affinity constant of 2.1x108 L/mol.The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work.Conclusions The modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.

  5. Downstream processing of monoclonal antibodies--application of platform approaches. (United States)

    Shukla, Abhinav A; Hubbard, Brian; Tressel, Tim; Guhan, Sam; Low, Duncan


    This paper presents an overview of large-scale downstream processing of monoclonal antibodies and Fc fusion proteins (mAbs). This therapeutic modality has become increasingly important with the recent approval of several drugs from this product class for a range of critical illnesses. Taking advantage of the biochemical similarities in this product class, several templated purification schemes have emerged in the literature. In our experience, significant biochemical differences and the variety of challenges to downstream purification make the use of a completely generic downstream process impractical. Here, we describe the key elements of a flexible, generic downstream process platform for mAbs that we have adopted at Amgen. This platform consists of a well-defined sequence of unit operations with most operating parameters being pre-defined and a small subset of parameters requiring development effort. The platform hinges on the successful use of Protein A chromatography as a highly selective capture step for the process. Key elements of each type of unit operation are discussed along with data from 14 mAbs that have undergone process development. Aspects that can be readily templated as well as those that require focused development effort are identified for each unit operation. A brief description of process characterization and validation activities for these molecules is also provided. Finally, future directions in mAb processing are summarized.

  6. Pharmacokinetics of biotech drugs: peptides, proteins and monoclonal antibodies. (United States)

    Lin, Jiunn H


    With the advances in recombinant DNA biotechnology, molecular biology and immunology, the number of biotech drugs, including peptides, proteins and monoclonal antibodies, available for clinical use has dramatically increased in recent years. Although pharmacokinetic principles are equally applicable to the large molecule drugs and conventional small molecule drugs, the underlying mechanisms for the processes of absorption, distribution, metabolism and excretion (ADME) of large molecule drugs are often very different from that of small molecule drugs. Therefore, a good understanding of the ADME processes of large molecule drugs is essential in support of the development of therapeutic biologics. The purpose of this article is to review the current knowledge of the ADME processes that govern the pharmacokinetics of biotech drugs. The challenges encountered by orally administered peptide and protein drugs, and the nature of lymphatic absorption after subcutaneous administration will be discussed. In addition, molecular mechanisms of biodistribution, metabolism and renal excretion of biotech drugs will also be discussed. Finally, approaches used for prediction of human pharmacokinetics of protein drugs will be briefly discussed.


    Institute of Scientific and Technical Information of China (English)

    WANGMin-Yi; HEZhi-Ying; WANGHan-Zheng


    The purpose of this study is to investigate the mechanism by which antiprogcsterone monoclonal antibodies block early pregnancy in mice. The mechanism of passive immunization is a complex issue as indicated below:

  8. Purification of a Mycoplasma pneumoniae adhesin by monoclonal antibody affinity chromatography.


    Leith, D K; Baseman, J B


    A 165,000-dalton surface protein of Mycoplasma pneumoniae, designated protein P1, appears to be the major attachment ligand of the pathogen. We employed monoclonal antibody affinity chromatography to obtain purified protein P1.

  9. The role of radiolabeled anti-TNFα monoclonal antibodies for diagnostic purposes and therapy evaluation

    NARCIS (Netherlands)

    Glaudemans, A. W.J.M.; Dierckx, R. A.J.O.; Kallenberg, C. G.M.; Anzola Fuentes, K. L.


    Radiolabelled cytokines and monoclonal antibodies are an emerging class of radiopharmaceuticals for imaging inflammation. These radiopharmaceuticals bind to their targets with high affinity and specificity and therefore have excellent diagnostic potential for imaging of patients with chronic inflamm


    Institute of Scientific and Technical Information of China (English)



    Three monoclonal antibodies (MAbs) were raised against both pig and rabbit zona pellucida with a dual immunization protocol employing heat soluble pig zona (HSPZ) and heat soluble rabbit zona (HSRZ), Of the 140 wells screencd, 12 wells were positive to

  11. Real-time kinetic analysis applied to the production of bispecific monoclonal antibodies for radioimmunodetection of cancer. (United States)

    Horenstein, A L; Poiesi, C; DeMonte, L; Camagna, M; Mariani, M; Albertini, A; Malavasi, F


    An automated biosensor system designed for measuring molecular interactions in real-time and without labelling of the reactants has been used to evaluate the association/dissociation rate and affinity constants of bivalent monoclonal antibodies and a monovalent bispecific monoclonal antibody. Observed differences in affinity between parental and bispecific antibody produced were related to the association rate constants, since the dissociation rate constants were in the same range. Values were also closely related to radioimmunochemical data. These results indicate that the biosensor system, besides presenting several advantages for characterizing antigen-antibody interaction, is valuable for selecting monoclonal antibodies with properties which might be useful in the development of bispecific monoclonal antibodies.

  12. High Ki-67 Immunohistochemical Reactivity Correlates With Poor Prognosis in Bladder Carcinoma: A Comprehensive Meta-Analysis with 13,053 Patients Involved. (United States)

    Luo, Yihuan; Zhang, Xin; Mo, Meile; Tan, Zhong; Huang, Lanshan; Zhou, Hong; Wang, Chunqin; Wei, Fanglin; Qiu, Xiaohui; He, Rongquan; Chen, Gang


    Ki-67 is considered as one of prime biomarkers to reflect cell proliferation and immunohistochemical Ki-67 staining has been widely applied in clinical pathology. To solve the widespread controversy whether Ki-67 reactivity significantly predicts clinical prognosis of bladder carcinoma (BC), we performed a comprehensive meta-analysis by combining results from different literature. A comprehensive search was conducted in the Chinese databases of WanFang, China National Knowledge Infrastructure and Chinese VIP as well as English databases of