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Sample records for blood lymphocytes pbls

  1. Mutagenic and morphologic impacts of 1.8 GHz radiofrequency radiation on human peripheral blood lymphocytes (hPBLs) and possible protective role of pre-treatment with Ginkgo biloba (EGb 761)

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    Esmekaya, Meric Arda, E-mail: mericarda@yahoo.com [Department of Biophysics, Gazi University, Faculty of Medicine and Gazi Non-ionizing Radiation, Protection (GNRP) Center, Ankara (Turkey); Aytekin, Ebru [Department of Medical Genetics, Gazi University Faculty of Medicine, Ankara (Turkey); Ozgur, Elcin; Gueler, Goeknur [Department of Biophysics, Gazi University, Faculty of Medicine and Gazi Non-ionizing Radiation, Protection (GNRP) Center, Ankara (Turkey); Ergun, Mehmet Ali [Department of Medical Genetics, Gazi University Faculty of Medicine, Ankara (Turkey); Oemeroglu, Suna [Department of Histology and Embryology, Gazi University, Faculty of Medicine, Ankara (Turkey); Seyhan, Nesrin [Department of Biophysics, Gazi University, Faculty of Medicine and Gazi Non-ionizing Radiation, Protection (GNRP) Center, Ankara (Turkey)

    2011-12-01

    The mutagenic and morphologic effects of 1.8 GHz Global System for Mobile Communications (GSM) modulated RF (radiofrequency) radiation alone and in combination with Ginkgo biloba (EGb 761) pre-treatment in human peripheral blood lymphocytes (hPBLs) were investigated in this study using Sister Chromatid Exchange (SCE) and electron microscopy. Cell viability was assessed with 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) reduction assay. The lymphocyte cultures were exposed to GSM modulated RF radiation at 1.8 GHz for 6, 8, 24 and 48 h with and without EGb 761. We observed morphological changes in pulse-modulated RF radiated lymphocytes. Longer exposure periods led to destruction of organelle and nucleus structures. Chromatin change and the loss of mitochondrial crista occurred in cells exposed to RF for 8 h and 24 h and were more pronounced in cells exposed for 48 h. Cytoplasmic lysis and destruction of membrane integrity of cells and nuclei were also seen in 48 h RF exposed cells. There was a significant increase (p < 0.05) in SCE frequency in RF exposed lymphocytes compared to sham controls. EGb 761 pre-treatment significantly decreased SCE from RF radiation. RF radiation also inhibited cell viability in a time dependent manner. The inhibitory effects of RF radiation on the growth of lymphoctes were marked in longer exposure periods. EGb 761 pre-treatment significantly increased cell viability in RF + EGb 761 treated groups at 8 and 24 h when compared to RF exposed groups alone. The results of our study showed that RF radiation affects cell morphology, increases SCE and inhibits cell proliferation. However, EGb 761 has a protective role against RF induced mutagenity. We concluded that RF radiation induces chromosomal damage in hPBLs but this damage may be reduced by EGb 761 pre-treatment. - Highlights: Black-Right-Pointing-Pointer RF Radiation inhibits cell proliferation. Black-Right-Pointing-Pointer RF radiation induces chromosomal damage

  2. Monocyte-derived dendritic cells enhance cell proliferation and porcine circovirus type 2 replication in concanavalin A-stimulated swine peripheral blood lymphocytes in vitro.

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    Lin, Chun-Ming; Jeng, Chian-Ren; Hsiao, Shih-Hsuan; Lee, Yao; Tsai, Yi-Chieh; Chia, Mi-Yuan; Pang, Victor Fei

    2012-01-15

    Dendritic cells (DCs) are professional antigen presenting cells cooperating with other immune cells for the activation of innate and adaptive immune responses. The objective of the present study was to investigate the replication activity of porcine circovirus type 2 (PCV2) in DCs and/or lymphocytes during their cross talk and its possible mechanism. Two models were set, herein. Swine blood monocyte (Mo)-derived DCs (MoDCs) or peripheral blood lymphocytes (PBLs) were inoculated with PCV2 prior to their co-cultivation. Bacterial lipopolysaccharide (LPS) and concanavalin A (Con A) were used to stimulate MoDCs and PBLs, respectively. During 6 days of cultivation, a high PCV2 antigen-containing rate without detectable intranuclear signals and a slight but significant increase in the copy number of PCV2 genome were detected in PCV2-inoculated MoDCs. The presence of LPS alone or PCV2-free PBLs, however, had no effect on the location of PCV2 antigens or copy number of PCV2 genome in PCV2-inoculated MoDCs. On the contrary, active PCV2 replication occurred in Con A-stimulated PCV2-inoculated PBLs. When compared with blood Mos, MoDCs induced significantly higher cell proliferation and intensified PCV2 replication in Con A-stimulated PCV2-inoculated PBLs, for which direct contact between MoDCs and lymphocytes was required. Among the cytokines secreted by Con A-activated PBLs, interleukin (IL)-2, but not IL-4 or interferon-γ, could induce cell proliferation and PCV2 replication in PCV2-inoculated PBLs. The findings suggest that although MoDCs support only limited PCV2 replication in themselves, their accessory cell function is required for cell proliferation and PCV2 replication in PCV2-infected lymphocytes.

  3. Molybdate modulates mitogen and cyclosporin responses of human peripheral blood lymphocytes.

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    Michelis, Fotios V; Delitheos, Andreas; Tiligada, Ekaterini

    2011-07-01

    The trace element molybdenum (Mo) is an essential component of key physiological systems in animals, plants and microorganisms. The molybdate oxoanion MoO(4)(2-) has been demonstrated to cause diverse yet poorly understood biochemical and pharmacological effects, such as non-specific inhibition of phosphatases and stabilization of steroid receptors. This study aimed to investigate the effects of molybdate on the activation of human peripheral blood lymphocytes (hPBLs) ex vivo and its potential interaction with the widely used immunosuppressant drug cyclosporin A (CsA). Lymphocyte activation was evaluated by performing multiple experiments determining blastogenesis in cultured peripheral blood lymphocytes obtained from 5 healthy volunteers, following stimulation induced by phytohemagglutinin (PHA), in the absence or presence of 0.05-10 mM sodium molybdate or/and 2.5-30 μg/mL CsA. Blastogenesis was assessed by a morphometric assay based on the relative proportions of unactivated lymphocytes, activated lymphoblasts and cells with aberrant morphology after PHA-induced activation. Molybdate concentrations up to 1 mM showed no effect on lymphocyte blastogenesis, while higher concentrations exerted immunosuppressive actions on cultured hPBLs. Co-administration of 0.1 mM sodium molybdate with CsA, at doses up to 20 μg/mL, induced no alteration in the response of cultured hPBLs to CsA. However, molybdate potentiated the immunosuppressive action of higher CsA concentrations, implying a likely dose-related synergistic interaction of the two agents in PHA-stimulated blood lymphocytes. These observations are indicative of the possible biological importance of molybdate oxoanions in the modulation of hPBL activation that may have pharmacological consequences during the therapeutic application of immunomodulatory drugs.

  4. DNA damage in peripheral blood lymphocytes in patients during combined chemotherapy for breast cancer

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    Sanchez-Suarez, Patricia [Oncological Research Unit, Oncology Hospital, National Medical Center S-XXI, Instituto Mexicano del Seguro Social (IMSS), Av. Cuauhtemoc 330, Col. Doctores, 06725 Mexico, D.F. (Mexico); Ostrosky-Wegman, Patricia [Biomedical Research Institute, Universidad Nacional Autonoma de Mexico (UNAM), Mexico City (Mexico); Gallegos-Hernandez, Francisco [Department of Clinical Oncology, Oncology Hospital, National Medical Center S-XXI, Instituto Mexicano del Seguro Social (IMSS), Mexico City (Mexico); Penarroja-Flores, Rubicelia; Toledo-Garcia, Jorge [Oncological Research Unit, Oncology Hospital, National Medical Center S-XXI, Instituto Mexicano del Seguro Social (IMSS), Av. Cuauhtemoc 330, Col. Doctores, 06725 Mexico, D.F. (Mexico); Bravo, Jose Luis [Atmospheric Sciences Institute, Universidad Nacional Autonoma de Mexico (UNAM), Mexico City (Mexico); Rojas del Castillo, Emilio [Biomedical Research Institute, Universidad Nacional Autonoma de Mexico (UNAM), Mexico City (Mexico); Benitez-Bribiesca, Luis [Oncological Research Unit, Oncology Hospital, National Medical Center S-XXI, Instituto Mexicano del Seguro Social (IMSS), Av. Cuauhtemoc 330, Col. Doctores, 06725 Mexico, D.F. (Mexico)], E-mail: luisbenbri@mexis.com

    2008-04-02

    Combined chemotherapy is used for the treatment of a number of malignancies such as breast cancer. The target of these antineoplastic agents is nuclear DNA, although it is not restricted to malignant cells. The aim of the present study was to assess DNA damage in peripheral blood lymphocytes (PBLs) of breast cancer patients subjected to combined adjuvant chemotherapy (5-fluorouracil, epirubicin and cyclophosphamide, FEC), using a modified comet assay to detect DNA single-strand breaks (SSB) and double-strand breaks (DSB). Forty-one female patients with advanced breast cancer before and after chemotherapy and 60 healthy females participated in the study. Alkaline and neutral comet assays were performed in PBLs according to a standard protocol, and DNA tail moment was measured by a computer-based image analysis system. Breast cancer patients before treatment had higher increased background levels of SSB and DSB as compared to healthy women. During treatment, a significant increase in DNA damage was observed after the 2nd cycle, which persisted until the end of treatment. Eighty days after the end of treatment the percentage of PBLs with SSB and DSB remained elevated, but the magnitude of DNA damage (tail moment) returned to baseline levels. There was no correlation between PBL DNA damage and response to chemotherapy. DNA-SSB and DSB in PBLs are present in cancer patients before treatment and increase significantly after combined chemotherapy. No correlation with response to adjuvant chemotherapy was found. Biomonitoring DNA damage in PBLs of cancer patients could help prevent secondary effects and the potential risks of developing secondary cancers.

  5. mRNA expression of dopamine receptors in peripheral blood lymphocytes of computer game addicts.

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    Vousooghi, Nasim; Zarei, Seyed Zeinolabedin; Sadat-Shirazi, Mitra-Sadat; Eghbali, Fatemeh; Zarrindast, Mohammad Reza

    2015-10-01

    Excessive playing of computer games like some other behaviors could lead to addiction. Addictive behaviors may induce their reinforcing effects through stimulation of the brain dopaminergic mesolimbic pathway. The status of dopamine receptors in the brain may be parallel to their homologous receptors in peripheral blood lymphocytes (PBLs). Here, we have investigated the mRNA expression of dopamine D3, D4 and D5 receptors in PBLs of computer game addicts (n = 20) in comparison to normal subjects (n = 20), using a real-time PCR method. The results showed that the expression level of D3 and D4 dopamine receptors in computer game addicts were not statistically different from the control group. However, the expression of the mRNA of D5 dopamine receptor was significantly down-regulated in PBLs of computer game addicts and reached 0.42 the amount of the control group. It is concluded that unlike with drug addiction, the expression levels of the D3 and D4 dopamine receptors in computer game addicts are not altered compared to the control group. However, reduced level of the D5 dopamine receptor in computer game addicts may serve as a peripheral marker in studies where the confounding effects of abused drugs are unwanted.

  6. Initiation but no execution - modulation of peripheral blood lymphocyte apoptosis in rheumatoid arthritis - a potential role for heat shock protein 70

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    Chuturgoon Anil A

    2011-11-01

    Full Text Available Abstract Background Rheumatoid arthritis (RA is a chronic autoimmune disease, which causes synovial damage. Persistence of lymphocyte infiltrates in the rheumatoid synovium has been attributed to abnormal apoptosis. While not comprehensively investigated, perturbations in peripheral blood lymphocyte (PBL apoptosis may also be involved in perpetuation of autoimmune processes in RA. Methods We investigated total, CD4+ and CD19+ PBL apoptosis in our study cohort by monitoring the translocation of phosphatidylserine using the Annexin-V assay. To examine the role of death receptor mediated apoptosis as well as activation-induced-cell-death (AICD, PBLs were labeled with CD95/Fas and CD69 markers and enumerated by flow cytometry. Proteolytic activity of initiator and executioner caspases was determined by luminometry. DNA fragmentation assays were used to examine whether apoptotic signals were transduced to the nucleus. Quantitative PCR arrays were used to investigate apoptotic pathways associated with RA-PBLs. Since heat-shock-protein-70 (HSP70 is an inducible protein which modulates apoptotic signals, we determined HSP70 levels by intra-cellular flow cytometry and western blots. Results The RA-PBLs showed signs of elevated apoptosis whilst in circulation. These include increases in the loss of plasma membrane asymmetry, indicated by increased externalization of phosphatidylserine (especially in B-lymphocytes. RA-PBLs showed a bias to CD95/Fas mediated apoptotic pathways, but low levels of the CD69 marker suggested that this was not associated with immune activation. Although downstream markers of apoptosis such as caspase-3/7 activity, were increased, no DNA fragmentation was observed in RA-PBLs. Interestingly, elevated levels of apoptosis did not correlate with absolute lymphocyte counts in RA patients. Levels of HSP70 were highly elevated in RA-PBLs compared to controls. Conclusion The results suggest that while apoptosis may be initiated in RA-PBLs

  7. Increased micronucleus, nucleoplasmic bridge, and nuclear bud frequencies in the peripheral blood lymphocytes of diesel engine exhaust-exposed workers.

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    Zhang, Xiao; Duan, Huawei; Gao, Feng; Li, Yuanyuan; Huang, Chuanfeng; Niu, Yong; Gao, Weimin; Yu, Shanfa; Zheng, Yuxin

    2015-02-01

    The International Agency for Research on Cancer has recently reclassified diesel engine exhaust (DEE) as a Group 1 carcinogen. Micronucleus (MN), nucleoplasmic bridge (NPB), and nuclear bud (NBUD) frequencies in peripheral blood lymphocytes (PBLs) are associated with cancer risk. However, the impact of DEE exposure on MN frequency has not been thoroughly elucidated due to mixed exposure and its impact on NPB and NBUD frequencies has never been explored in humans. We recruited 117 diesel engine testing workers with exclusive exposure to DEE and 112 non-DEE-exposed workers, and then we measured urinary levels of 4 mono-hydroxylated polycyclic aromatic hydrocarbons (OH-PAHs) using high-performance liquid chromatography-mass spectrometry as well as MN, NPB, and NBUD frequencies in PBLs using cytokinesis-block MN assay. The DEE-exposed workers exhibited significantly higher MN, NPB, and NBUD frequencies than the non-DEE-exposed workers (P frequencies (all P frequencies persisted in DEE-exposed workers (P = 0.001). The percent of MN frequencies increased, on average, by 23.99% (95% confidential interval, 9.64-39.93) per 1-unit increase in ln-transformed 9-OHPh. Our results clearly show that exposure to DEE can induce increases in MN, NPB, and NBUD frequencies in PBLs and suggest that DEE exposure level is associated with MN frequencies.

  8. AKRs expression in peripheral blood lymphocytes from smokers: the role of body mass index.

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    Barrón-Vivanco, B S; Rothenberg, S J; Medina-Díaz, I M; Robledo-Marenco, L; Rojas-García, A E; Hernández-Cadena, L; Poblete-Naredo, I; Elizondo, G; Albores, A

    2013-04-01

    Aldo-keto reductases (AKRs) metabolize a wide range of substrates, including polycyclic aromatic hydrocarbons (PAHs), generating metabolites (o-quinones) and reactive oxygen species (ROS), which are capable of initiating and promoting carcinogenesis. Exposure to PAHs, their metabolites, and ROS further increase AKRs isoform expression that may amplify oxidative damage. Human AKR enzymes are highly polymorphic, and allelic variants may contribute to different AKRs expression in individuals. Despite the importance of AKRs in PAHs metabolism, there are no studies that evaluate, in general human populations, the effect of PAHs on AKRs expression in peripheral blood lymphocytes (PBLs). The aim of this study was to determine the effect of tobacco smoke exposure, and AKR1A1*2 and AKR1C3*2 polymorphisms, on AKR1A1 and AKR1C1-AKR1C3 messenger RNA (mRNA) levels in PBLs from smokers. In the smoker group, there is a statistically significant positive association between AKR1A1, AKR1C1, and AKR1C3 mRNA induction and urine cotinine levels in individuals with a body mass index (BMI) less than 25. However, AKR1A1*2 and AKR1C3*2 alleles did not influence AKR1A1 and AKR1C1-AKR1C3 mRNA levels. These results suggest that AKRs induction by PAHs in smokers' PBLs is associated with BMI; therefore, the role of adipose tissue accumulation in PAHs' effects needs further investigation.

  9. Shorter telomere length in peripheral blood lymphocytes of workers exposed to polycyclic aromatic hydrocarbons.

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    Pavanello, Sofia; Pesatori, Angela-C; Dioni, Laura; Hoxha, Mirjam; Bollati, Valentina; Siwinska, Ewa; Mielzyńska, Danuta; Bolognesi, Claudia; Bertazzi, Pier-Alberto; Baccarelli, Andrea

    2010-02-01

    Shorter telomere length (TL) in peripheral blood lymphocytes (PBLs) is predictive of lung cancer risk. Polycyclic aromatic hydrocarbons (PAHs) are established lung carcinogens that cause chromosome instability. Whether PAH exposure and its molecular effects are linked with shorter TL has never been evaluated. In the present study, we investigated the effect of chronic exposure to PAHs on TL measured in PBLs of Polish male non-current smoking cokeoven workers and matched controls. PAH exposure and molecular effects were characterized using measures of internal dose (urinary 1-pyrenol), effective dose [anti-benzo[a]pyrene diolepoxide (anti-BPDE)-DNA adduct], genetic instability (micronuclei, MN) and DNA methylation [p53 promoter and Alu and long interspersed nuclear element-1 (LINE-1) repetitive elements, as surrogate measures of global methylation] in PBLs. TL was measured by real-time polymerase chain reaction. Cokeoven workers were heavily exposed to PAHs (79% exceeded the urinary 1-pyrenol biological exposure index) and exhibited lower TL (P = 0.038) than controls, as well as higher levels of genetic and chromosomal alterations [i.e. anti-BPDE-DNA adduct and MN (P < 0.0001)] and epigenetic changes [i.e. p53 gene-specific promoter and global methylation (P

  10. Increased mitochondrial DNA content in peripheral blood lymphocytes from HIV-infected patients with lipodystrophy.

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    Cossarizza, Andrea; Riva, Agostino; Pinti, Marcello; Ammannato, Silvia; Fedeli, Paolo; Mussini, Cristina; Esposito, Roberto; Galli, Massimo

    2003-08-01

    We have evaluated mitochondrial (mt) DNA content in CD4 and CD8 peripheral blood lymphocytes (PBLs) from HIV-infected patients taking highly active antiretroviral therapy (HAART) who display different types of adipose tissue alterations. A cross-sectional study was performed in a total of 23 patients with lipodystrophy (LD): nine patients with fat accumulation, six patients with fat loss, eight patients with combined form, who were compared to 11 individuals infected by HIV without LD (HIV-positive) and 10 seronegative controls (CTRL). PBLs were obtained by standard methods, that is, gradient density centrifugation on Ficoll, and CD4 or CD8 cells were positively isolated by magnetic sorting to eliminate the contamination of platelets. mtDNA content was then measured by an original assay based upon real-time PCR. mtDNA content was significantly increased in CD4 T cells from patients with LD, while no differences were present between CD4 and CD8 cells from HIV-positive and CTRL individuals. Nor were any differences found when comparing LD or HIV-positive patients treated with stavudine or zidovudine, or taking D-drugs or non D-drugs. Patients with fat accumulation had significantly higher mtDNA content compared to HIV-positive and CTRL, this phenomenon regarding both CD4 and CD8 PBLs. Considering all HIV-positive patients (including LD), mtDNA content showed a significant, positive correlation with cholesterolaemia but not with triglyceridaemia and glycaemia. Relatively high mtDNA content in LD patients, as well as the correlation between mtDNA content and cholesterol in all HIV-positive subjects, suggest the involvement of mitochondria in such a pathology. However, further studies are needed to confirm these initial observations and ascertain whether the quantification of mtDNA in PBL is a useful and reliable marker to investigate and monitor HAART-related changes in fat distribution.

  11. Use of recombinant lentivirus pseudotyped with vesicular stomatitis virus glycoprotein G for efficient generation of human anti-cancer chimeric T cells by transduction of human peripheral blood lymphocytes in vitro

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    Kolokoltsov Andrey A

    2006-02-01

    Full Text Available Abstract Background Genetic redirection of lymphocytes that have been genetically engineered to recognize antigens other than those originally programmed in their germlines is a potentially powerful tool for immunotherapy of cancers and potentially also of persistent viral infections. The basis for this procedure is that both cancers and some viruses have developed strikingly similar mechanisms of evading attacks by host immune mechanisms. To redirect human peripheral blood lymphocytes (PBLs with a chimeric T cell receptor (chTCR so that they recognize a new target requires a high degree of transfection efficiency, a process that is regarded as technically demanding. Results Infection with a retroviral vector carrying a chTCR cassette was shown to transduce 100% of rapidly dividing murine T cells but typically, only ~10% of PBLs could be infected with the same vector. In contrast with other retroviruses, lentiviruses integrate their genomes into non-dividing cells. To increase host cell range, vesicular stomatitis virus G protein was pseudotyped with a lentivirus vector, which resulted in ~100% PBL transduction efficiency. Signaling of PBLs bearing chimeric receptors was shown by specific proliferation on exposure to cells expressing cognate ligand. Further, T-bodies against CEA showed a startling abilty to cause regression of maligant colon tumors in a nude mouse model of human cancer. Conclusion A lentivirus/VSV pseudotyped virus, which does not require replicating cells for integration of its genome, efficiently transduced a high proportion of human PBLs with chTCRs against CEA. PBLs transduced by infection with a lentivirus/VSV pseudotyped vector were able to proliferate specifically in vitro on exposure to CEA-expressing cells and further they had a startling therapeutic effect in a mouse model of human colon cancer.

  12. 1. cap alpha. ,25-Dihydroxyvitamin D/sub 3/ inhibits. gamma. -interferon synthesis by normal human peripheral blood lymphocytes

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    Reichel, H.; Koeffler, H.P.; Tobler, A.; Norman, A.W.

    1987-05-01

    1..cap alpha..,25-Dihydroxyvitamin D/sub 3/ (1,25-(OH)/sub 2/D/sub 3/), the biologically active metabolite of vitamin D/sub 3/, inhibited synthesis of ..gamma..-interferon (IFN-..gamma..) by phytohemagglutinin-activated peripheral blood lymphocytes (PBLs). A significant reduction of IFN-..gamma.. protein levels in PBL culture medium was achieved with a physiologic 1,25-(OH)/sub 2/D/sub 3/ concentration, 1,25-(OH)/sub 2/D/sub 3/ also inhibited accumulation of IFN-..gamma.. mRNA in activated PBLs in a dose-dependent fashion. The ability of 1,25-(OH)/sub 2/D/sub 3/ to modulate IFN-..gamma.. protein synthesis was unaltered in the presence of high concentrations of recombinant human interleukin 2. The suppression of IFN-..gamma.. synthesis by PBLs was specific for 1,25-(OH)/sub 2/D/sub 3/; the potencies of other vitamin D/sub 3/ metabolites were correlated with their affinities for the cellular 1,25-(OH)/sub 2/D/sub 3/ receptor. The time course of 1,25-(OH)/sub 2/D/sub 3/ receptor expression in phytohemagglutinin-activated PBLs was correlated with the time course of 1,25-(OH)/sub 2/D/sub 3/-mediated inhibition of IFN-..gamma.. synthesis. Finally, the authors examined the effects of 1,25-(OH)/sub 2/D/sub 3/ on the constitutive IFN-..gamma.. production by two human T-lymphocyte lines transformed by human T-lymphotropic virus type I. The cell lines were established from a normal donor (cell line S-LB1) and from a patient with vitamin D-dependent rickets type 2 (cell line Ab-VDR). IFN-..gamma.. synthesis by S-LB1 cells was inhibited in a dose-dependent fashion by 1,25-(OH)/sub 2/D/sub 3/, whereas IFN-..gamma.. synthesis by Ab-VDR cells was not altered by 1,25-(OH)/sub 2/D/sub 3/. The data presented in this study provide evidence for a role of 1,25-(OH)/sub 2/D/sub 3/ in immunoregulation.

  13. Combination of p53(ser15) and p21/p21(thr145) in peripheral blood lymphocytes as potential Alzheimer's disease biomarkers.

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    Tan, Mengshan; Wang, Shuying; Song, Juexian; Jia, Jianping

    2012-05-16

    Alzheimer's disease (AD) is still difficult to be precisely diagnosed in its early stage to date. Establishing of reliable and manageable disease-specific biological markers is required to improve diagnostic accuracy. Based on the hypothesis of cell cycle regulatory failure at the early stage of AD, we tested whether cell cycle regulating proteins p53, p21 and their phosphorylated forms p53(ser15), p21(thr145) were changed in AD patients and whether these proteins could be used as diagnostic biomarkers. Western bolt, Enzyme-linked immunosorbent assay (ELISA), immunofluorescent staining and flow cytometry (FCM) analysis were employed to analyze levels of these proteins in peripheral blood lymphocytes (PBLs) from 95 controls, 94 AD, 12 Parkinson's disease (PD) and 15 vascular dementia (VaD) patients. Compared with controls, p53(ser15) and p21(thr145) levels were significantly increased and p21 level was significantly decreased in PBLs of AD patients but not in PD or VaD, while p53 was increased in both AD and VaD patients. The receiver operating characteristic (ROC) curve analysis showed that the specificity and sensitivity were 76% and 84% for p53, 88% and 82% for p53(ser15), 80% and 75% for p21 and 84% and 68% for p21(thr145) in identifying AD patients. The relatively high diagnostic accuracy support these proteins, especially p53(ser15) and p21 in PBLs may become potential biomarkers for diagnosis of AD.

  14. Effect of antilymphoma antibody, 131I-Lym-1, on peripheral blood lymphocytes in patients with non-Hodgkin's lymphoma.

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    Schillaci, Orazio; DeNardo, Gerald L; DeNardo, Sally J; Goldstein, Desiree S; Kroger, Linda A; O'Donnell, Robert T; Lamborn, Kathleen R

    2007-08-01

    Anti-CD20 monoclonal antibodies (mAbs), unlabeled rituximab (Rituxan, Biogen Idec Inc., Cambridge, MA; and Genentech Inc., South San Francisco, CA) or radiolabeled 90Y-ibritumomab (Zevalin, Biogen Idec Inc., Cambridge, MA) and 131I-tositumomab (Bexxar; Glaxo Smith Kline, Research Triangle Park, NC), have proven to be effective therapy for non-Hodgkin's lymphoma (NHL), but also induce immediate and persistent decreases in normal peripheral blood lymphocytes (PBLs). Lym-1, a mAb that selectively targets malignant lymphocytes, also has induced therapeutic responses and prolonged survival in patients with NHL when labeled with iodine-131 (131I). We have retrospectively examined its effect on PBLs in 41 NHL patients that had received 131I-Lym-1 therapy. Absolute lymphocyte counts (ALCs) were evaluated before and after the first and last 131I-Lym-1 infusion. Modest decreases in PBLs were observed in most of the patients. Using strict criteria to define recovery, time to recovery was determined for 19 patients, with the remainder censored because of insufficient follow-up (median follow up for censored patients: 22 days). Using Kaplan-Meier estimates, it would be predicted that 31% of patients would recover by 28 days and that median time to recovery would be 44 days after the last 131I-Lym-1 infusion. No predictors were found for time to recovery, considering such factors as the administered Lym-1 or 131I dose, spleen volume, or radiation doses to the body, marrow, or spleen. The data suggest that the effect of 131I-Lym-1 on ALC is the result of a nonspecific radiation effect, rather than a specific Lym-1 mAb effect. The shorter time required for ALC recovery after 131I-Lym-1 when compared to that reported for anti-CD20 mAbs, whether radiolabeled or otherwise, is probably related to differing mechanisms for lymphocytotoxicity and lesser Lym-1 antigenic density on normal B-lymphocytes.

  15. Integration Analysis of MicroRNA and mRNA Expression Profiles in Human Peripheral Blood Lymphocytes Cultured in Modeled Microgravity

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    C. Girardi

    2014-01-01

    Full Text Available We analyzed miRNA and mRNA expression profiles in human peripheral blood lymphocytes (PBLs incubated in microgravity condition, simulated by a ground-based rotating wall vessel (RWV bioreactor. Our results show that 42 miRNAs were differentially expressed in MMG-incubated PBLs compared with 1 g incubated ones. Among these, miR-9-5p, miR-9-3p, miR-155-5p, miR-150-3p, and miR-378-3p were the most dysregulated. To improve the detection of functional miRNA-mRNA pairs, we performed gene expression profiles on the same samples assayed for miRNA profiling and we integrated miRNA and mRNA expression data. The functional classification of miRNA-correlated genes evidenced significant enrichment in the biological processes of immune/inflammatory response, signal transduction, regulation of response to stress, regulation of programmed cell death, and regulation of cell proliferation. We identified the correlation of miR-9-3p, miR-155-5p, miR-150-3p, and miR-378-3p expression with that of genes involved in immune/inflammatory response (e.g., IFNG and IL17F, apoptosis (e.g., PDCD4 and PTEN, and cell proliferation (e.g., NKX3-1 and GADD45A. Experimental assays of cell viability and apoptosis induction validated the results obtained by bioinformatics analyses demonstrating that in human PBLs the exposure to reduced gravitational force increases the frequency of apoptosis and decreases cell proliferation.

  16. Alteration of peripheral blood lymphocyte subsets in acute pancreatitis

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    Miroslawa Pietruczuk; Milena I Dabrowska; Urszula Wereszczynska-Siemiatkowska; Andrzej Dabrowski

    2006-01-01

    AIM: To evaluate peripheral blood lymphocyte subsets in patients with acute pancreatitis (AP).METHODS: Twenty patients with mild AP (M-AP) and 15 with severe AP (S-AP) were included in our study. Peripheral blood lymphocytes were examined at d 1-3, 5,10 and 30 by means of flow cytometry.RESULTS: A significant depletion of circulating lymphocytes was found in AP. In the early AP, the magnitude of depletion was similar for T- and B- lymphocytes. In the late course of S-AP, B-lymphocytes were much more depleted than T-lymphocytes. At d 10, strong shift in the CD7+/CD19+ ratio implicating predominance of Tover B-lymphocytes in S-AP was found. Among T-lymphocytes, the significant depletion of the CD4+ population was observed in M-AP and S-AP, while CD8+ cells were in the normal range. Lymphocytes were found to strongly express activation markers: CD69, CD25, CD28,CD38 and CD122. Serum interleukin-2 (IL-2), IL-4, IL-5,IL-10, interferon-γ (IFN-γ) and tumor necrosis factor-α(TNF-α) levels were significantly increased in both forms of AP. The magnitude of elevation of cytokines known to be produced by Th2 was much higher than cytokines produced by Th1 cells.CONCLUSION: AP in humans is characterized by significant reduction of peripheral blood T- and B-lymphocytes.

  17. Persistent expression and function of P-glycoprotein on peripheral blood lymphocytes identifies corticosteroid resistance in patients with systemic lupus erythematosus.

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    Kansal, Amit; Tripathi, Deepak; Rai, Mohit K; Agarwal, Vikas

    2016-02-01

    Corticosteroids (CS) are the mainstay of treatment in systemic lupus erythematosus (SLE) patients. However, some patients have poor response to CS treatment. Among the multiple mechanisms of CS resistance, overexpression of P-glycoprotein (P-gp) on peripheral blood lymphocytes (PBL) may be one of them as this result in efflux of CS from lymphocytes. Thus, we evaluated the role of P-gp protein on PBLs in patients with SLE in its response to CS therapy. SLE patients (n = 42) (fulfilling ACR revised criteria) who were naïve to CS and immunosuppressive drugs were enrolled. Disease activity was assessed using SLE disease activity index (SLEDAI) and expression, and function of P-gp was evaluated by flow cytometry at baseline and after 3 months of therapy with CS. At 3 months, patients with SLEDAI >4 and SLEDAI ≤4 were grouped as nonresponders and responders, respectively. P-gp expression was significantly increased on PBLs of SLE patients as compared to healthy controls (p < 0.001). P-gp expression and function correlated with SLEDAI (r = 0.49, p = 0.005; and r = 0.49, p = 0.001, respectively). P-gp expression and function were not different in responders and nonresponders at baseline. However, at 3 months of CS therapy, P-gp expression and function decreased in responders (p < 0.001 and p < 0.005, respectively), whereas in nonresponders, it remained unchanged. Persistent overexpression and activity of P-gp are associated with poor response to CS in CS naïve patients of SLE.

  18. A variable CD3⁺ T-cell frequency in peripheral blood lymphocytes associated with type 1 diabetes mellitus development in the LEW.1AR1-iddm rat.

    Directory of Open Access Journals (Sweden)

    Tanja Arndt

    Full Text Available PURPOSE: The LEW.1AR1-iddm rat is an animal model of human type 1 diabetes mellitus (T1DM, which arose through a spontaneous mutation within the MHC-congenic inbred strain LEW.1AR1 (RT1(r². In contrast to the diabetes-resistant LEW.1AR1 background strain in LEW.1AR1-iddm rats a highly variable T-cell frequency could be observed in peripheral blood lymphocytes (PBLs. METHODS: In this study we therefore characterised the T-cell repertoire within the PBLs of the two strains by flow cytometry analysis and identified the CD3⁺ T-cell phenotype and its possible linkage to diabetes susceptibility. To map loci conferring susceptibility to variable CD3⁺ T-cell frequency, backcross strains (N2 were generated with the genetically divergent BN and PAR rats for microsatellite analysis. RESULTS: The LEW.1AR1-iddm rat strain was characterised by a higher variability of CD3⁺ T-cells in PBLs along with a slightly decreased mean value compared to the LEW.1AR1 background strain. The reason for this reduction was a decrease in the CD4⁺ T-cell count while the CD8⁺ T-cell proportion remained unchanged. However, both T-cell subpopulations showed a high variability. This resulted in a lower CD4⁺/CD8⁺ T-cell ratio than in LEW.1AR1 rats. Like LEW.1AR1-iddm rats all animals of the backcross populations, N2 BN and N2 PAR rats, also showed large variations of the CD3⁺ T-cell frequency. The phenotype of variable CD3⁺ T-cell frequency mapped to the telomeric region of chromosome 1 (RNO1, which is identical with the already known Iddm8 diabetes susceptibility region. The data indicate that a variable CD3⁺ T-cell frequency in PBLs is genetically linked to diabetes susceptibility in the LEW.1AR1-iddm rat. CONCLUSION: The T-cell variability in PBLs could be related to the previously reported imbalance between regulatory and effector T-cell populations which results in beta-cell autoimmunity. Since similar T-cell phenotypes have also been described in human T1

  19. Chromosomal instability in the lymphocytes of breast cancer patients

    Directory of Open Access Journals (Sweden)

    Harsimran Kaur

    2009-01-01

    Full Text Available Genomic instability in the tumor tissue has been correlated with tumor progression. In the present study, chromosomal aberrations (CAs in peripheral blood lymphocytes (PBLs of breast tumor patients were studied to assess whether chromosomal instability (CIN in PBLs correlates with aggressiveness of breast tumor (i.e., disease stage and has any prognostic utility. Cultured blood lymphocyte metaphases were scored for aberrations in 31 breast cancer patients and 20 healthy age and sex-matched controls. A variety of CAs, including aneuploidy, polyploidy, terminal deletions, acentric fragments, double minutes, chromatid separations, ring chromosome, marker chromosome, chromatid gaps, and breaks were seen in PBLs of the patients. The CAs in patients were higher than in controls. A comparison of the frequency of metaphases with aberrations by grouping the patients according to the stage of advancement of disease did not reveal any consistent pattern of variation in lymphocytic CIN. Neither was any specific chromosomal abnormality found to be associated with the stage of cancer. This might be indicative of the fact that cancer patients have constitutional CIN, which predisposes them to the disease, and this inherent difference in the level of genomic instability might play a role in disease progression and response to treatment.

  20. Lymphocyte subset reference intervals in blood donors from northeastern Brazil

    Directory of Open Access Journals (Sweden)

    ALEX J.L. TORRES

    2015-06-01

    Full Text Available The reference intervals for leukocytes and lymphocytes currently used by most clinical laboratories present limitations as they are primarily derived from individuals of North American and European origin. The objective this study was to determine reference values for peripheral blood B lymphocytes, T lymphocyte subsets (CD4+, CD8+, naïve, memory, regulatory, TCRαβ and TCRγδ+ and NK cells from blood donors in Salvador-Bahia, Brazil. Results: The proportion of included male subjects was 73.7% and the median ages of males (34 and females (35 were found to be similar. Absolute counts total lymphocytes subsets to both gender was 1,956 (1,060-4,186 cells and relative values 34%. The T CD4+ and T CD8+ lymphocytes relative values was 51% (20-62 and 24% (9-28, respectively. The most statistically significant finding observed was a higher percentage of B lymphocytes (p=0.03 in females. Commonly cited subset reference intervals were found to be consistent with values in several populations from different geographic areas.

  1. Characterization of two heat shock proteins (Hsp70/Hsc70) from grass carp (Ctenopharyngodon idella): evidence for their differential gene expression, protein synthesis and secretion in LPS-challenged peripheral blood lymphocytes.

    Science.gov (United States)

    Zhang, Anying; Zhou, Xiaofei; Wang, Xinyan; Zhou, Hong

    2011-06-01

    Two cDNAs, encoding the stress-inducible 70-kDa heat shock protein (Hsp70) and the constitutively expressed 70-kDa heat shock cognate protein (Hsc70), were isolated from grass carp. The Hsp70 and Hsc70 cDNAs were 2250 bp and 2449 bp in length and contained 1932 bp and 1953 bp open reading frames, respectively. Tissue distribution results showed that Hsp70/Hsc70 was highly expressed in gill, kidney, head kidney and peripheral blood lymphocytes (PBLs). Using grass carp PBLs as a cell model, effects of lipopolysaccharide (LPS) on the mRNA and protein levels of Hsp70/Hsc70 were examined. In this case, LPS increased the mRNA expression of Hsp70 in a time- and dose-dependent manner, but had no effect on Hsc70 mRNA expression. In agreement with this, LPS elevated the intracellular Hsp70 markedly, but not the Hsc70 protein levels in parallel experiments. Furthermore, Hsp70 protein was also detected in culture medium. Moreover, inhibition of LPS on Hsp70 release in a time-dependent manner was observed, indicating that there may be a dynamic balance between Hsp70 stores and Hsp70 release in grass carp PBLs following exposure to LPS. Taken together, these results not only shed new insights into the different regulations of LPS on Hsp70/Hsc70 gene expression, protein synthesis and release, but also provide a basis for further study on the functional role of Hsp70 in fish immune response.

  2. [239Pu and chromosomal aberrations in human peripheral blood lymphocytes].

    Science.gov (United States)

    Okladnikova, N D; Osovets, S V; Kudriavtseva, T I

    2009-01-01

    The genome status in somatic cells was assessed using the chromosomal aberration (CA) test in peripheral blood lymphocytes from 194 plutonium workers exposed to occupational radiation mainly from low-transportable compounds of airborne 230Pu. Pu body burden at the time of cytogenetic study varied from values close to the method sensitivity to values multiply exceeding the permissible level. Standard (routine) methods of peripheral blood lymphocytes cultivation were applied. Chromatid- and chromosomal-type structural changes were estimated. Aberrations were estimated per 100 examined metaphase cells. The quantitative relationship between the CA frequency and Pu body burden and the absorbed dose to the lung was found. Mathematical processing of results was carried out based on the phenomenological model. The results were shown as theoretical and experimental curves. The threshold of the CA yield was 0.43 +/- 0.03 kBq (Pu body burden) and 6.12 +/- 1.20 cGy (absorbed dose to the lung).

  3. Radiosensitivity of peripheral blood lymphocytes in autoimmune disease

    Energy Technology Data Exchange (ETDEWEB)

    Harris, G. (Kennedy Inst. of Rheumatology, London (UK). Div. of Experimental Pathology); Cramp, W.A.; Edwards, J.C.; George, A.M.; Sabovljev, S.A.; Hart, L.; Hughes, G.R.V. (Hammersmith Hospital, London (UK)); Denman, A.M. (Northwich Park Hospital, Harrow (UK)); Yatvin, M.B. (Wisconsin Clinical Cancer Center, Madison (USA))

    1985-06-01

    The proliferation of peripheral blood lymphocytes, cultured with Con A, can be inhibited by ionizing radiation. Lymphocytes from patients with conditions associated with autoimmunity, such as rheumatoid arthritis, systemic lupus erythematosus and polymyositis, are more radiosensitive than those from healthy volunteers or patients with conditions not associated with autoimmunity. Nuclear material isolated from the lymphocytes of patients with autoimmune diseases is, on average, lighter in density than the nuclear material from most healthy controls. This difference in density is not related to increased sensitivity to ionizing radiation but the degree of post-irradiation change in density (lightening) is proportional to the initial density, i.e. more dense nuclear material always shows a greater upward shift after radiation. The recovery of pre-irradiation density of nuclear material, 1 h after radiation exposure, taken as an indication of DNA repair, correlates with the radiosensitivity of lymphocyte proliferation (Con A response); failure to return to pre-irradiation density being associated with increased sensitivity of proliferative response. These results require extension but, taken with previously reported studied of the effects of DNA methylating agents, support the idea that DNA damage and its defective repair could be important in the aetio-pathogenesis of autoimmune disease.

  4. Induction of micronuclei in human and mouse lymphocytes irradiated with gamma radiation and effect of panax ginseng C. A. Meyer

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Sung Ho; Oh, Heon; Lee, Song Eun [Chonnam National Univ., Kwangju (Korea, Republic of); Lee, Yun Sil; Kim, Tae Hwan [Korea Cancer Center Hospital, Seoul (Korea, Republic of); Jeong, Kyu Sik [Korea Research Institute of Bioscience and Biotechnology, Taejon (Korea, Republic of); Ryu, Si Yun [Chungnam National Univ., Taejon (Korea, Republic of)

    1997-09-01

    The frequencies of {gamma}-ray-induced micronuclei (MN) in Cytokinesis-Blocked (CB) lymphocytes at several doses were measured in three donors of human and C57BL/6 mice. Measurements performed after irradiation showed a dose-related increases in MN frequency in each of the donors studied. The relative sensitivity of mouse in Spleen Lymphocytes (SLs) compared with human Peripheral Blood Lymphocytes (PBLs) was estimated by best fitting linear-quadratic model based on the radiation-induced MN data over the range from 0 cGy to 400 cGy. In the case of MN frequency with 0.2 per CB cell, the relative sensitivity of mouse SLs was 1.67. Compared with the radiation-induced MN formation in the PBLs of human, the SLs of mouse were more radiosensitive. Using this MN assay with human PBLs and mouse SLs, studies were performed to determine whether the water fraction of ginseng (Panax ginseng C.A.Meyer)against radiation-induced MN in human PBLs after in vitro irradiation (3Gy) and in SLs of C57BL/6 mice after in vivo irradiation (3Gy). The frequency of MN in human PBLs was reduced by water fraction of ginseng (0.5mg/ml of medium) both pre-and post treatment (p<0.01) in vitro. In addition, the frequency of MN in mouse SLs was also reduced by pretreatment of ginseng (2mg/ml of drinking water for 7 days) in vivo.

  5. Peripheral blood lymphocytes DNA in patients with chronic liver diseases

    Institute of Scientific and Technical Information of China (English)

    Vasiliy I Reshetnyak; Tatyana I Sharafanova; Ludmila U Ilchenko; Elena V Golovanova; Gennadiy G Poroshenko

    2001-01-01

    BACKGROUND Viral replication in blood cells with nucleuses may lead to the damage of lymphocytes genetic apparatus and the beginning of immunopathological reactions.AIM Of this investigation is to reveal the damage to peripheral blood lymphocytes (PBL)DNA in the patients with chronic liver diseases.MATERIALS AND METHODS Sixteen-ninepatients with chronic liver diseases (37 patients with chronic viral hepatitis, 2 patients with liver cirrhosis of mixed etiology (alcohol + virus G),30 women with primary biliary cirrhosis-PBC)were examined. The condition of DNA structure of PBL-was measured by the fluorescenceanalysis of DNA unwinding (FADU) technique with modification. Changes of fluorescence (in %) reflected the DNA distractions degree (thepresence of DNA single-stranded breaks and alkalinelabile sights).RESULTS AND CONCLUSION . The quantity of DNA single-stranded breaks and alkalinelabile sightsin DNA in all patients with chronic viral hepatitis .didnt differ from the control group,excluding the patients with chronic hepatitis (CH) C + G. Patients with HGV and TTV monoinfection had demonstrated the increase of the DNA single-stranded breaks PBL quantity.This fact may be connected with hypothesisabout the viruses replication in white blood cells discussed in the literature. Tendency to increase quantity of DNA PBL damages in the patients with primary biliary cirrhosis (PBC) accordingly to the alkaline phosphatase activity increase was revealed. Significant decrease of the DNA single-stranded breaks and alkalinelabile sights in the PBC patients that were treated with prednison was demonstrated. Probably, the tendency to increase the quantity of DNA singlestranded breaks and alkalinelabile sights in lymphocytes of the PBC patients was depended on the surplus of the blood bile acid content.

  6. In-vitro carbofuran induced genotoxicity in human lymphocytes and its mitigation by vitamins C and E.

    Science.gov (United States)

    Sharma, Ratnesh Kumar; Sharma, Bechan

    2012-01-01

    Various efforts have been made in past in order to predict the underlying mechanism of pesticide-induced toxicity using in vitro and animal models, however, these predictions may or may not be directly correlated with humans. The present study was designed to investigate the carbofuran induced genotoxicity and its amelioration by vitamins C and E by treating human peripheral blood lymphocytes (PBLs) with different concentrations (0, 0.5, 1.25, 2.5, 3.75 and 5.0 μM) of this compound. The treatment of PBLs with carbofuran displayed significant DNA damage in concentration dependent manner. The carbofuran induced genotoxicity could be ameliorated to considerable extent by pretreatment of PBLs with equimolar (10 μM) concentration of each of the vitamins C and E; the magnitude of protection by vitamin E being higher than by vitamin C. Also, it was found that the level of protection by these vitamins was higher when PBLs were treated with lower concentrations of pesticide. The significant DNA damage as observed by H_{2}O_{2}, a positive control in the present study, and its amelioration by natural antioxidants (vitamins C and E) lend an evidence to suggest that carbofuran would have caused genotoxicity via pesticide induced oxidative stress.

  7. In-Vitro Carbofuran Induced Genotoxicity in Human Lymphocytes and Its Mitigation by Vitamins C and E

    Directory of Open Access Journals (Sweden)

    Ratnesh Kumar Sharma

    2012-01-01

    Full Text Available Various efforts have been made in past in order to predict the underlying mechanism of pesticide-induced toxicity using in vitro and animal models, however, these predictions may or may not be directly correlated with humans. The present study was designed to investigate the carbofuran induced genotoxicity and its amelioration by vitamins C and E by treating human peripheral blood lymphocytes (PBLs with different concentrations (0, 0.5, 1.25, 2.5, 3.75 and 5.0 μM of this compound. The treatment of PBLs with carbofuran displayed significant DNA damage in concentration dependent manner. The carbofuran induced genotoxicity could be ameliorated to considerable extent by pretreatment of PBLs with equimolar (10 μM concentration of each of the vitamins C and E; the magnitude of protection by vitamin E being higher than by vitamin C. Also, it was found that the level of protection by these vitamins was higher when PBLs were treated with lower concentrations of pesticide. The significant DNA damage as observed by H2O2, a positive control in the present study, and its amelioration by natural antioxidants (vitamins C and E lend an evidence to suggest that carbofuran would have caused genotoxicity via pesticide induced oxidative stress.

  8. [Dynamics of blood lymphocyte blast-transformation parameters during hemosorption in patients with gastric ulcer].

    Science.gov (United States)

    Zhidkov, K P; Fedorova, L A; Polevshchikov, A V; Nazarov, P G

    1992-03-01

    The influence of autotransfusions of hemosorbent-treated blood on parameters of DNA-synthetic activity of blood lymphocytes was studied in 114 patients with gastric ulcer. A statistically significant increase in parameters of DNA-synthetic activity of lymphocytes was recorded simultaneously with the appearance of morphological signs of blast transformation on electrograms. A conclusion has been made on association of lymphocyte blastogenesis under the influence of autotransfusions with the process of ulcerous sanagenesis acceleration.

  9. Human T cell priming assay: depletion of peripheral blood lymphocytes in CD25(+) cells improves the in vitro detection of weak allergen-specific T cells.

    Science.gov (United States)

    Vocanson, Marc; Achachi, Amine; Mutez, Virginie; Cluzel-Tailhardat, Magalie; Varlet, Béatrice Le; Rozières, Aurore; Fournier, Philippe; Nicolas, Jean-François

    2014-01-01

    To develop an in vitro assay that recapitulates the key event of allergic contact dermatitis (ACD), that is the priming of effector T cells by hapten-presenting dendritic cells, and then allows for the sensitive detection of chemical allergens represents a major challenge. Classical human T cell priming assays (hTCPA) that have been developed in the past, using hapten-loaded monocyte-derived dendritic cells (MDDCs) as antigen-presenting cells and peripheral blood lymphocytes (PBLs) as responding cells, were not efficient to prime T cells to common allergens with moderate/weak sensitizing properties. Recent progress in the understanding of the effector and regulatory mechanisms of ACD have shown that T cell priming requires efficient uptake of allergens by immunogenic DCs and that it is controlled by several subsets of regulatory cells including CD25(+) Tregs. We therefore analyzed various parameters involved in allergen-specific T cell activation in vitro and showed that priming of allergen-specific T cells is hampered by several subsets of immune cells comprising CD1a(neg) DCs, CD25(+) T cells, and CD56(+) regulatory cells.CD4(+)CD25(+)FoxP3(+) Tregs prevented the in vitro T cell priming to moderate/weak allergens, and depletion of human PBLs in CD25(+) cells significantly increased specific T cell proliferation and IFN-γ secretion. CD56(+) cells exerted an additional control of T cell priming since co-depletion of both CD56(+) and CD25(+) cells improved the magnitude of chemical-specific T cell activation. Finally, CD1a(low) MDDCs were able to inhibit T cell activation obtained by allergen-pulsed CD1a(high) MDDC. Moreover, we showed that uptake by DC of allergen-encapsulated nanoparticles significantly increased their activation status and their ability to prompt specific T cell activation. Hence, by combining the different strategies, i.e., depletion of CD25(+) and CD56(+) cells, use of CD1a(high) MDDC, and nanoparticle encapsulation of allergens, it was

  10. Chromosome Aberration in Human Blood Lymphocytes Exposed to Energetic Protons

    Science.gov (United States)

    Hada, M.; George, Kerry A.; Cucinotta, F. A.

    2008-01-01

    During space flight, astronauts are exposed to a space radiation consisting of high-energy protons, high charge and energy (HZE) nuclei, as well as secondary particles that are generated when the primary particles penetrate the spacecraft shielding. Secondary particles have a higher LET value than primary protons and therefore expected to have a higher relative biological effectiveness (RBE). To investigate this theory, we exposed human peripheral blood lymphocytes to protons with energies of 250 MeV, 800MeV, 2 GeV, or 2.5 GeV. LET values for these protons ranged from 0.4 to 0.2 keV/micrometer. and doses ranged from 0.2 to 3 Gy. Over this energy the probability of nuclear reaction leading to secondary radiation, and the multiplicity of reaction produces such as neutrons and mesons increases substantially. The effect of aluminum and polyethylene shielding was also assessed using the 2 GeV and 2.5GeV proton beams. After exposure lymphocytes were stimulated to divide and chromosomes were collected from cells in the first G2 and metaphase cell cycle after exposure using a chemical induced premature chromosome condensation (PCC) technique. Dose response data for chromosome damage was analyzed using the fluorescence in situ hybridization (FISH) chromosome painting technique. Selected samples were also analyzed with multicolor FISH (mFISH) and multicolor banding FISH (mBAND) techniques. Data indicates that the dose response for simple-type exchanges is similar for proton and gamma exposure, whereas protons induce higher yields of complex exchanges that are LET dependent. RBE values will be presented for each proton energy, and the effects of shielding and possible cytogenetic signatures of proton exposure will be discussed.

  11. Chromosome aberrations in human blood lymphocytes exposed to energetic protons

    Science.gov (United States)

    Hada, Megumi; George, Ms Kerry; Cucinotta, Francis A.

    During space flight, astronauts are exposed to space radiation consisting of high-energy protons, high charge and energy (HZE) nuclei, as well as secondary particles that are generated when the primary particles penetrate the spacecraft shielding. Secondary particles have a higher LET value than primary protons and are therefore expected to have a higher relative biological effectiveness (RBE). To investigate this theory, we exposed human peripheral blood lymphocytes to protons with energies of 250 MeV, 800MeV, 2 GeV, or 2.5 GeV. LET values for these protons ranged from 0.4 to 0.2 keV/µm. and doses ranged from 0.2 to 3 Gy. Over this energy range the probability of nuclear reaction leading to secondary radiation, and the multiplicity of reaction products such as neutrons and mesons increases substantially. The effect of aluminum and polyethylene shielding was also assessed using the 2 GeV and 2.5GeV proton beams. After exposure lymphocytes were stimulated to divide and chromosomes were collected from cells in the first G2 and metaphase cell cycle after exposure using a chemical induced premature chromosome condensation (PCC) technique. Dose response data for chromosome damage was analyzed using the fluorescence in situ hybridization (FISH) chromosome painting technique. Selected samples were also analyzed with multicolor FISH (mFISH) and multicolor banding FISH (mBAND) techniques. Data indicates that the dose response for simple-type exchanges is similar for proton and gamma exposure, whereas protons induce higher yields of complex exchanges that are energy dependent. RBE values will be presented for each proton energy, and the effects of shielding and possible cytogenetic signatures of proton exposure will be discussed.

  12. Total lymphoid irradiation in multiple sclerosis: blood lymphocytes and clinical course

    Energy Technology Data Exchange (ETDEWEB)

    Cook, S.D.; Devereux, C.; Troiano, R.; Zito, G.; Hafstein, M.; Lavenhar, M.; Hernandez, E.; Dowling, P.C.

    1987-11-01

    We have found a significant relationship between blood lymphocyte count and prognosis in 45 patients receiving either total lymphoid irradiation or sham irradiation for chronic progressive multiple sclerosis. Patients with sustained lymphocyte counts less than 900 mm-3 for prolonged periods after treatment showed less rapid progression over the ensuing 3 years than did patients with multiple sclerosis who had lymphocyte counts above this level (p less than 0.01). Our results suggest that a simple laboratory test, the absolute blood lymphocyte count, may serve as a valuable barometer for monitoring the amount of immunosuppressive therapy needed to prevent progression in patients with multiple sclerosis, and possibly other autoimmune diseases.

  13. CLONAL CHRONIC LYMPHOCYTIC LEUKEMIA-LIKE B-LYMPHOCYTES IN THE BLOOD OF PATIENTS WITH CUTANEOUS T-CELL DISORDERS

    NARCIS (Netherlands)

    DAENEN, S; VADER, PCV; BLOM, N; PIETENS, J; HOLLEMA, H; SMIT, JW

    1993-01-01

    A population of B cells with characteristics of chronic lymphocytic leukaemia was found in the peripheral blood of four patients who presented with cutaneous infiltration of atypical CD4+ T cells with cerebriform nuclei. The B cells had a low density of immunoglobulin on their surface membrane, expr

  14. 早、中、晚孕期胎盘因子对人外周血淋巴细胞CD4,CCR5和CXCR4表达的影响%Effect of first, second, and third trimester placental factorson CD4, CCR5,and CXCR4 expression in human peripheral blood lymphocytes

    Institute of Scientific and Technical Information of China (English)

    李莉平; 康佳丽; 夏薇; 曾耀英

    2008-01-01

    Objective To investigate effect of first, second, and third trimester placental factors (PF) on CD4, CCR5, and CXCR4 expression in human peripheral blood lymphocytes (PBLs), and to explore their influence on human immunodeficiency virus (HIV) vertical transmission.Methods Human peripheral blood mononuclear cells (PBMCs) were treated with first, second,and third trimester PF (concentration 25%) respectively for 24 hours. The expression of CD4, CCR5,and CXCR4 in PBLs, and the percentages of CCR5+, CXCR4+,and CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes were determined with flow cytometry.Results All trimester PFs reduced CCR5 expression in PBLs. The efficiency of the first trimester PF was higher than that of the second and third trimester PF. The percentage of CCR5+ cells in peripheral blood CD4+ lymphocytes of PF groups was significantly lower than that of the control group, and the percentage of CCR5+ cells in peripheral blood CD4+ lymphocytes of the first trimester PF group was significantly lower than that of the second and third trimester group. The percentages of CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes of PF groups were significantly decreased as compared with the control group, and the percentage of CCR5+CXCR4+ cells in peripheral blood CD4+ lymphocytes of the first trimester PF group was significantly lower than that of the third trimester PF group.Conclusion PF can reduce the expression of CCR5 in human PBLs and peripheral blood CD4+ lymphocytes, indicating that PF might reduce R5 virus infection via preventing HIV entry, and might play an important role in reducing R5 virus intrauterine infection.%目的:通过研究早、中、晚孕期胎盘因子(PF)对人外周血淋巴细胞(PBLs)中CD4, CCR5和CXCR4表达的作用,探讨PF在人免疫缺陷病毒-1 (HIV-1)垂直传播中的作用及其机制.方法:制备早、中、晚孕期PF.分离人外周血单个核细胞,并分别与相对浓度为25%的早

  15. In vitro susceptibilities in lymphocytes from mothers and cord blood to the monofunctional alkylating agent EMS

    DEFF Research Database (Denmark)

    Wyatt, N P; Falque-Gonzalez, C; Farrar, D;

    2007-01-01

    at the Bradford Royal Infirmary collected venous blood samples from mothers at the time of birth and venous cord blood post-delivery. Lymphocytes were isolated from both blood types and examined in the alkaline comet assay using the monofunctional alkylating agent ethyl methanesulphonate (EMS). There were...

  16. Sodium homeostasis in lymphocytes and blood pressure alterations before and during salt restriction in normotensives and in essential hypertensives

    DEFF Research Database (Denmark)

    Jest, P; Pedersen, K E; Klitgaard, N A

    1986-01-01

    Blood pressure, lymphocytic sodium content and sodium efflux were studied in hypertensive and normotensive subjects during salt restriction. Diastolic blood pressure decreased significantly in both groups. In essential hypertension the initial high lymphocyte sodium content decreased during salt...... mechanisms with regard to lymphocyte sodium metabolism differs between hypertensive and normotensive subjects....

  17. Research on spontaneously emerged chromosomal aberrations in the periphery blood lymphocytes in cattle ('Busa' breed).

    Science.gov (United States)

    Hasanbasić, Danica; Rukavina, Dunja; Hodzić, Aida; Brka, Muhamed; Vegara, Mensur; Hamamdzić, Muhidin

    2007-11-01

    Knowledge of spontaneous aberrations, namely, of their frequency in non-irradiated cells is of paramount importance not only in cytogenetic research, but also in contemporary animal production. The paper deals with research on spontaneously emerged chromosomal aberrations in the peripheral blood lymphocytes in the cattle of 'Busa' breed. To obtain metaphase chromosomes the conventional method of lymphocyte cultivation was used, albeit slightly modified and adapted to the examined animals and the laboratory conditions. The research findings indicate that a certain percent of spontaneously emerged chromosomal aberrations of chromatid type (gap and break) have been found in the peripheral blood lymphocytes in the cattle of 'Busa' breed.

  18. Cerebrolysin administration reduces oxidative stress-induced apoptosis in lymphocytes from healthy individuals.

    Science.gov (United States)

    Formichi, Patrizia; Radi, Elena; Battisti, Carla; Di Maio, Giuseppe; Muresanu, Dafin; Federico, Antonio

    2012-11-01

    Cerebrolysin is the only drug available for clinical use containing active fragments of some important neurotrophic factors obtained from purified porcine brain proteins, which has long been used for the treatment of dementia and stroke sequels. Cerebrolysin has growth factor-like activities and promotes neuronal survival and sprouting, however, its molecular mechanism still needs to be determined. It has been shown that Cerebrolysin may interact with proteolytic pathways linked to apoptosis. Administration of Cerebrolysin significantly reduces the number of apoptotic neurons after glutamate exposure. Furthermore, it has been reported that Cerebrolysin inhibits free radicals formation and lipid peroxidation. In vitro we evaluated the protective effects of Cerebrolysin towards spontaneous and induced apoptotic death in cells from healthy individuals. Peripheral blood lymphocytes (PBLs) from 10 individuals were used as cell model; 2-deoxy-D-ribose (dRib), a highly reducing sugar, was used as paradigm pro-apoptotic stimulus. Apoptosis was analysed using flow cytometry and fluorescence microscopy. Our results showed that Cerebrolysin significantly reduced the number of apoptotic PBLs after dRib treatment, although it had no significative effects on cells cultured in standard conditions. Our work showed a protective effect of Cerebrolysin on oxidative stress-induced apoptosis and suggested that PBLs can be used as an easy obtainable and handy cell model to verify Cerebrolysin effects in neurodegenerative pathologies.

  19. Classical scrapie prions in ovine blood are associated with B lymphocytes and platelet-rich plasma

    Directory of Open Access Journals (Sweden)

    Dassanayake Rohana P

    2011-11-01

    Full Text Available Abstract Background Classical scrapie is a naturally occurring transmissible spongiform encephalopathy of sheep and goats characterized by cellular accumulation of abnormal isoforms of prion protein (PrPSc in the central nervous system and the follicles of peripheral lymphoid tissues. Previous studies have shown that the whole blood and buffy coat blood fraction of scrapie infected sheep harbor prion infectivity. Although PrPSc has been detected in peripheral blood mononuclear cells (PBMCs, plasma, and more recently within a subpopulation of B lymphocytes, the infectivity status of these cells and plasma in sheep remains unknown. Therefore, the objective of this study was to determine whether circulating PBMCs, B lymphocytes and platelets from classical scrapie infected sheep harbor prion infectivity using a sheep bioassay. Results Serial rectal mucosal biopsy and immunohistochemistry were used to detect preclinical infection in lambs transfused with whole blood or blood cell fractions from preclinical or clinical scrapie infected sheep. PrPSc immunolabeling was detected in antemortem rectal and postmortem lymphoid tissues from recipient lambs receiving PBMCs (15/15, CD72+ B lymphocytes (3/3, CD21+ B lymphocytes (3/3 or platelet-rich plasma (2/3 fractions. As expected, whole blood (11/13 and buffy coat (5/5 recipients showed positive PrPSc labeling in lymphoid follicles. However, at 549 days post-transfusion, PrPSc was not detected in rectal or other lymphoid tissues in three sheep receiving platelet-poor plasma fraction. Conclusions Prion infectivity was detected in circulating PBMCs, CD72+ pan B lymphocytes, the CD21+ subpopulation of B lymphocytes and platelet-rich plasma of classical scrapie infected sheep using a sheep bioassay. Combining platelets with B lymphocytes might enhance PrPSc detection levels in blood samples.

  20. Human cord blood lymphocytes. Ultrastructural and immunologic surface marker characteristics: a comparison with B- and T-cell lymphomas

    Energy Technology Data Exchange (ETDEWEB)

    Hamburg, A.; Brynes, R.K.; Reese, C.; Golomb, H.M.

    1976-01-01

    The ultrastructural and surface marker characteristics of human cord blood lymphocytes were studied. These properties were compared with those in cells of patients in the leukemic phase of both malignant lymphoma, poorly differentiated lymphocytic type, and mycosis fungoides. Nuclear folding in cord blood lymphocytes was similar to that seen in lymphocytes of patients with malignant lymphoma, poorly differentiated lymphocytic type and mycosis fungoides. Surface marker characteristics of cord blood lymphocytes included increased percentages of surface IgD on cells bearing surface immunoglobulins and decreased percentages of E-rosette-forming cells. The hypothesis that both malignant lymphoma, poorly differentiated lymphocytic type and mycosis fungoides represent an arrest in the normal lymphocyte maturation sequence is discussed.

  1. The peripheral blood compartment in patients with Graves' disease: activated T lymphocytes and increased transitional and pre-naive mature B lymphocytes

    Science.gov (United States)

    Van der Weerd, K; Van Hagen, P M; Schrijver, B; Kwekkeboom, D J; De Herder, W W; Ten Broek, M R J; Postema, P T E; Van Dongen, J J M; Staal, F J T; Dik, W A

    2013-01-01

    Graves' disease (GD) is an autoimmune disease that involves aberrant B and T lymphocyte responses. Detailed knowledge about lymphocyte subpopulation composition will therefore enhance our understanding of the pathogenesis of GD and might support the development of new immunomodulatory treatment approaches. The aim of this study was to gain detailed insight into the composition of the peripheral blood lymphocyte compartment in GD before and during anti-thyroid drug therapy. Major B and T lymphocyte subpopulations were investigated by flow cytometry in peripheral blood from newly diagnosed GD patients (n = 5), GD patients treated with anti-thyroid drugs (n = 4), patients with recurrent GD (n = 7) and healthy controls (HC; n = 10). In GD patients, numbers of activated T lymphocytes [human leucocyte antigen D-related (HLA-DR)+ and CD25+] were increased. The B lymphocyte compartment in GD was characterized by significantly higher numbers of transitional (CD38highCD27−, P < 0·03) and pre-naive mature (CD38lowCD27−IgD+CD5+, P < 0·04) B lymphocytes, while memory populations were slightly decreased. The increased numbers of CD5+, transitional and pre-naive mature B lymphocytes correlated positively with fT4 plasma levels. GD is associated with increased numbers of activated T lymphocytes and transitional and pre-naive mature CD5+ B lymphocytes within the peripheral blood. The increase in CD5+ B lymphocytes was due mainly to an increase in transitional and pre-naive mature B lymphocytes. Increased fT4 plasma levels might be associated with this increase in transitional and pre-naive mature CD5+ B lymphocytes. PMID:23901889

  2. EFFECTS OF INTERFERON THERAPY UPON IMMUNE MARKER PROFILE AND ENZYMATIC ACTIVITY IN PERIPHERAL BLOOD LYMPHOCYTES OF PATIENTS WITH RENAL CANCER

    Directory of Open Access Journals (Sweden)

    L. M. Kurtasova

    2014-01-01

    Full Text Available We have observed forty-four patients with metastatic renal cancer before and after interferon therapy. Immune markers of of peripheral blood lymphocytes were determined by flow cytometry. Activity of NAD (P-dependent dehydrogenase in blood lymphocytes was studied by means of bioluminescence technique. Changes of immune marker profiles and enzymatic activities of peripheral blood lymphocytes were found in patients with renal cancer after a course of interferon therapy.

  3. Immunophenotyping of Lymphocyte T and B in the Peripheral Blood of Systemic Lupus Erythematosus

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The immunophenotyping expression levels of lymphocyte in the peripheral blood from 21 patients with active systemic lupus erythematosus (SLE) were analyzed by using the immunofluorescence labeling-flow cytometry technique to investigate the immunophenotyping expression of lymphocytes T and B in the peripheral blood of active SLE patients and its clinical value. It was showed that, compared with normal controls, the expression of CD+3, CD+4 and the ratio of CD+4/CD+8 in the peripheral blood of these patients were decreased (P<0.01), while the expression of CD+8, CD+20 was significantly increased (P<0.01). It was suggested that both T and B cells in patients with active SLE involved in immunoregulation, were activated. The abnormal expression of lymphocyte immunophenotyping could influence the immune reaction in SLE patients, which might be one of the important pathogenesis factors in SLE.

  4. Glutamate decreases the secretion of IL-10 by peripheral blood lymphocytes in persons with autoimmune thyroiditis.

    Science.gov (United States)

    Kvaratskhelia, E; Dabrundashvili, N; Gagua, M; Maisuradze, E; Mikeladze, D

    2008-11-01

    Human T lymphocytes expose ionotropic and metabotropic glutamate receptors, which control immune responses, cell activation, maturation, and death. Several cytokines release during inflammation which identification may have important physiological and clinical implications. Main biological function of IL-10 is limitation and termination of inflammatory responses and the regulation of differentiation and proliferation of several immune cells. Various inflammatory molecules regulated the secretion of IL-8 and IL-10, but the action of glutamate on the biosynthesis of cytokines is unknown. We have found that in peripheral blood lymphocytes glutamate at the concentrations within normal plasma levels (1 x 10(-5) M), as well as at lower concentration (0.3 x 10(-6) M) changes the secretion of immunosuppressive cytokine IL-10, whereas synthesis of proinflammatory chemokine, IL-8 did not changed significantly. Moreover, our results have shown that peripheral blood lymphocytes from patients with autoimmune thyroiditis release less IL-10 at both concentration of glutamate than peripheral blood lymphocytes from healthy persons. These data suggest that glutamate decrease the secretion of IL-10 by peripheral blood lymphocytes, especially in patients with autoimmune thyroiditis that may be responsible for prolongation of inflammation.

  5. Effect of oral salt loading on blood pressure and lymphocyte sodium metabolism in borderline hypertension

    DEFF Research Database (Denmark)

    Pedersen, K E; Jest, P; Klitgaard, N A

    1986-01-01

    A randomized double-blind cross-over trial was performed to test the effects of oral salt loading (normal diet + 200 mmol NaCl/day for 4 weeks followed by normal diet + 400 mmol/day for 1 week) against placebo on blood pressure and lymphocyte sodium homeostasis in 10 young borderline hypertensive...... men, genetically predisposed for essential hypertension. Salt loading caused no significant changes in blood pressure levels, lymphocyte sodium content and efflux. In conclusion, our subjects seem insensitive to a few weeks of excessive salt intake....

  6. Granzyme release and caspase activation in activated human T-lymphocytes.

    Science.gov (United States)

    Zapata, J M; Takahashi, R; Salvesen, G S; Reed, J C

    1998-03-20

    Recently it has been reported that caspase-3 activation occurs in stimulated T-lymphocytes without associated apoptosis (Miossec, C., Dutilleul, V., Fassy, F., and Diu-Hercend, A. (1997) J. Biol. Chem. 272, 13459-13462). To explore this phenomenon, human peripheral blood lymphocytes (PBLs) were stimulated with mitogenic lectins or anti-CD3 antibody, and the proteolytic processing of different caspases and caspase substrates was analyzed by immunoblotting. Proteolytic processing of caspases-3 and -7 and the caspase substrates poly(ADP-ribose) polymerase, GDP dissociation inhibitor, and PKCdelta was observed when PBLs were activated in vitro, and lysates were prepared using RIPA buffer which contains 1% Nonidet P-40, 0.5% deoxycholate, and 0.1% SDS. In contrast, when a lysis buffer containing 2% SDS was used, the caspases remained in their zymogen pro-forms, and no proteolytic processing of caspase substrates was detected. Moreover, in experiments using intact cells and a cell-permeable fluorigenic caspase substrate, no caspase activity was observed in activated T-cells, whereas it was clearly detected when PBLs were treated with the apoptosis-inducing anticancer drug etoposide. Since the granzyme B is a direct activator of caspase-3 and its expression is induced following T-cell activation, we tested the effects of anti-GraB, an engineered serpin that specifically inhibits GraB. When the activated T-lymphocytes were lysed in RIPA buffer containing anti-GraB, no proteolytic processing or activation of caspase-3 was observed, strongly suggesting that release of GraB or similar proteases from their storage sites in cytotoxic granules during the lysis procedure is responsible for caspase activation. These findings demonstrate that T-cells do not process caspases upon activation and caution about the method of cell lysis used when studying granzyme-expressing cells.

  7. 2-Methoxyestradiol induce the conversion of human peripheral blood memory B lymphocytes into plasma cells.

    Science.gov (United States)

    Cayer, Marie-Pierre; Drouin, Mathieu; Proulx, Maryse; Jung, Daniel

    2010-04-15

    2-Methoxyestradiol (2ME), an end-metabolite of 17beta-estradiol, is an antiproliferative agent that is currently being tested in clinical trials for cancer treatment. We hereby report that sub-cytotoxic concentrations of 2ME influence the in vitro proliferation of human peripheral blood B lymphocytes. More surprisingly, we have observed that 2ME induces the conversion of CD138(-) B lymphocytes into CD138(+) cells of phenotype similar to immunoglobulin (Ig)-secreting plasma cells. Normal human B lymphocytes expressing CD138 increased in response to 2ME in a dose-dependent fashion, from 2% at baseline up to 31% in cells cultured in the presence of 0.75 microM 2ME. Moreover, most of the converted cells were also CD27(+) and secreted high levels of IgG (151 microg/10(6)cells/24h). IEF studies revealed that conversion occurred in a polyclonal manner. We then exploited this effect of 2ME to gain further insights into the molecular mechanisms that govern changes in transcription factors involved in plasma cells differentiation. Plasma cells generated by 2ME treatment of normal human B lymphocytes expressed elevated levels of IRF4 and reduced levels of Pax5 and Bcl-6. Similarly, levels of XBP-1 and Blimp-1 transcripts were increased. Our results suggest that the differentiation of peripheral blood B lymphocytes into plasma cells requires a similar modulation of transcription factors expression that for tonsil and bone marrow B lymphocytes.

  8. Isolation of labile Fcgamma-receptors from human peripheral blood lymphocytes and production of an antiserum.

    Science.gov (United States)

    Sandilands, G P; Peel, M G; Froebel, K S; Belch, J J; MacSween, R N

    1985-05-01

    In this study, we have isolated membranelabile Fcgamma-receptors (i.e. FcgammaR I) from normal human peripheral blood lymphocytes and have produced a rabbit antiserum to this protein. Using this antiserum, we have shown that membrane-labile and membrane-stable (i.e. FcgammaR II) Fcgamma-receptors are antigenically distinct and that these two forms of the receptors probably coexist on the same lymphocyte subpopulation. Moreover, it was apparent that lymphocyte FcgammaR Is are distinct from FcgammaRs expressed on other cell types (e.g. monocytes, polymorphs and spermatozoa). Preliminary evidence does suggest, however, that human platelets express an FcgammaR which is antigenically similar to human lymphocyte FcgammaR I.

  9. Studying the proliferation of human peripheral blood T lymphocytes in serum-free medium.

    Science.gov (United States)

    Tabakov, V U; Litvina, M M; Schepkina, J V; Jarilin, A A; Chestkov, V V

    2009-01-01

    We compared the cultivation of human peripheral blood lymphocytes in serum-free medium Hybris-2 and RPMI 1640 medium with 10% fetal bovine serum in the presence of phytohemagglutinin and interleukin-2. The optimal concentration of phytohemagglutinin significantly differed in serum-free and serum-containing media (0.5 and 5 microg/ml, [corrected] respectively). Both mitogens were more potent in stimulating the proliferation of lymphocytes in serum-free medium than in serum-containing medium. Strong proliferation of CD3(+) and CD4(+) T lymphocytes was observed in both media. The dynamics of other markers was similar in serum-free and serum-containing media. However, significant differences were revealed between individual donors. Our results indicate that the developed serum-free medium may be used in lymphocyte cultivation for scientific, diagnostic, and therapeutic purposes.

  10. Suppressed peripheral blood lymphocyte blastogenesis in pre- and postpartal sheep by chronic heat-stress, and suppressive property of heat-stressed sheep serum on lymphocytes.

    Science.gov (United States)

    Niwano, Y; Becker, B A; Mitra, R; Caldwell, C W; Abdalla, E B; Johnson, H D

    1990-01-01

    Phytohemagglutinin (PHA) and concanavalin A (Con A)-induced blastogenesis of peripheral blood lymphocytes was examined in heat-stressed pre- and postpartal sheep. The peak responses of lymphocytes to PHA and Con A in heat-stressed sheep revealed significant reduction before and after parturition compared with those in the corresponding control animals kept under thermoneutral conditions. Furthermore, the effect of serum from control or heat-stressed sheep on PHA-induced lymphocyte blastogenesis was examined. Supplementation of serum from heat-stressed sheep significantly suppressed the blastogenesis of lymphocytes obtained from healthy sheep, bovine, and human donors. Unlike dexamethasone, heat-stressed sheep serum did not inhibit IL-2 production by PHA-stimulated human peripheral blood lymphocytes. These results indicate that the immunosuppression of heat-stressed sheep is in part mediated by serum factor(s) that can modulate T-cell function in a species nonspecific manner.

  11. Identification of biomarkers for cervical cancer in peripheral blood lymphocytes using oligonucleotide microarrays

    Institute of Scientific and Technical Information of China (English)

    SHENG Jie; ZHANG Wei-yuan

    2010-01-01

    Background Oligonucleotide microarrays are increasingly being used to identify gene expression profiles that associated with complex genetic diseases. Peripheral lymphocytes communicate with cells and extracellular matrixes in almost all tissues and organs in human body, suggesting that the gene expression profiles in peripheral lymphocytes may reflect the presence of disease in the body. This study aimed to identify molecular biomarkers for cervical cancer in peripheral blood lymphocytes by using oligonucleotide microarrays.Methods Total RNA was extracted from peripheral blood lymphocytes of 24 early stage cervical cancer patients and 18 healthy controls. We used 22K Human Genome microarrays to profile peripheral blood lymphocytes from 4 early stage cervical cancer patients and compared their gene expression profiles with those from 3 healthy controls. Differentially expressed genes would be identified if they had adjusted P values of less than 0.05 and a groupwise average fold change greater than 1.5 or less than 0.67. Then the selected 5 genes were validated in the remaining 20 early stage cervical cancer patients and the 15 healthy controls by using real-time reverse-transcription polymerase chain reaction (RT-PCR).Results Genes identified by the gene selection program expressed differently between the blood samples of the early stage cervical cancer patients and those of the healthy controls. To validate the gene expression data, 5 genes were analyzed by real-time RT-PCR. In three of the 5 identified genes, tenasin-c (TNC), nuceolin (NCL), and enolase 2 (ENO2) showed a significant up-regulation in the blood samples of the early stage cervical cancer patients versus that of the healthy controls.Conclusions The up-regulation of TNC, NCL, and ENO2 in peripheral blood may be used to identify novel blood biomarkers for detecting cervical cancer in a clinically accessible surrogate tissue, and thus to provide a possibility to develop a noninvasive and predictive

  12. Peripheral Blood Lymphocyte Depletion After Hepatic Arterial {sup 90}Yttrium Microsphere Therapy for Hepatocellular Carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Carr, Brian I., E-mail: brianicarr@hotmail.com [Department of Surgery, University of Pittsburgh Medical Center, Pittsburgh, PA and Department of Nutrition and Exptl Biology, Saverio De Bellis Medical Research Institute, Castellana Grotte, Bari (Italy); Metes, Diana M. [Department of Surgery, University of Pittsburgh Medical Center, Pittsburgh, PA and Department of Nutrition and Exptl Biology, Saverio De Bellis Medical Research Institute, Castellana Grotte, Bari (Italy)

    2012-03-01

    Purpose: The short- and long-term effects of {sup 90}Yttrium microspheres therapy for hepatocellular carcinoma (HCC) on peripheral blood lymphocytes are unknown and were therefore examined. Methods and Materials: Ninety-two HCC patients were enrolled in a {sup 90}Yttrium therapy study and routine blood counts were examined as part of standard clinical monitoring. Results: We found an early, profound, and prolonged lymphopenia. In a subsequent cohort of 25 additional HCC patients, prospective flow cytometric immune-monitoring analysis was performed to identify specific changes on distinct lymphocyte subsets (i.e., CD3, CD4, CD8 T, and CD19 B lymphocytes) and NK cells absolute numbers, in addition to the granulocytes and platelets subsets. We found that the pretreatment lymphocyte subset absolute numbers (with the exception of NK cells) had a tendency to be lower compared with healthy control values, but no significant differences were detected between groups. Posttherapy follow-up revealed that overall, all lymphocyte subsets, except for NK cells, were significantly (>50% from pretherapy values), promptly (as early as 24 h) and persistently (up to 30 months) depleted post-{sup 90}Yttrium microspheres therapy. In contrast, granulocytes increased rapidly (24 h) to compensate for lymphocyte depletion, and remained increased at 1-year after therapy. We further stratified patients into two groups, according to survival at 1 year. We found that lack of recovery of CD19, CD3, CD8, and especially CD4 T cells was linked to poor patient survival. No fungal or bacterial infections were noted during the 30-month follow-up period. Conclusions: The results show that lymphocytes (and not granulocytes, platelets, or NK cells) are sensitive to hepatic arterial {sup 90}Yttrium without associated clinical toxicity, and lack of lymphocyte recovery (possibly leading to dysregulation of adaptive cellular immunity) posttherapy indicates poor survival.

  13. Evaluation of radioinduced damage and repair capacity in blood lymphocytes of breast cancer patients

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    P.A. Nascimento

    2001-02-01

    Full Text Available Genetic damage caused by ionizing radiation and repair capacity of blood lymphocytes from 3 breast cancer patients and 3 healthy donors were investigated using the comet assay. The comets were analyzed by two parameters: comet tail length and visual classification. Blood samples from the donors were irradiated in vitro with a 60Co source at a dose rate of 0.722 Gy/min, with a dose range of 0.2 to 4.0 Gy and analyzed immediately after the procedure and 3 and 24 h later. The basal level of damage and the radioinduced damage were higher in lymphocytes from breast cancer patients than in lymphocytes from healthy donors. The radioinduced damage showed that the two groups had a similar response when analyzed immediately after the irradiations. Therefore, while the healthy donors presented a considerable reduction of damage after 3 h, the patients had a higher residual damage even 24 h after exposure. The repair capacity of blood lymphocytes from the patients was slower than that of lymphocytes from healthy donors. The possible influence of age, disease stage and mutations in the BRCA1 and BRCA2 genes are discussed. Both parameters adopted proved to be sensitive and reproducible: the dose-response curves for DNA migration can be used not only for the analysis of cellular response but also for monitoring therapeutic interventions. Lymphocytes from the breast cancer patients presented an initial radiosensitivity similar to that of healthy subjects but a deficient repair mechanism made them more vulnerable to the genotoxic action of ionizing radiation. However, since lymphocytes from only 3 patients and 3 normal subjects were analyzed in the present paper, additional donors will be necessary for a more accurate evaluation.

  14. Two small lymphocyte subpopulations in human peripheral blood. I. Purification and surface marker profiles

    DEFF Research Database (Denmark)

    Hokland, M; Hokland, P; Heron, I

    1978-01-01

    By means of simple rosette sedimentation methods two subsets from human peripheral blood lymphocytes have been isolated: (1) (E, Fc)- and (2) (E, Ig)-. The first subset was obtained by centrifuging suspensions of macrophage-depleted PBL in which E and EA rosettes had been allowed to form simultan......By means of simple rosette sedimentation methods two subsets from human peripheral blood lymphocytes have been isolated: (1) (E, Fc)- and (2) (E, Ig)-. The first subset was obtained by centrifuging suspensions of macrophage-depleted PBL in which E and EA rosettes had been allowed to form...... of small subsets of human lymphocytes are effective and easy to perform and might be used to purify cells for functional studies. Udgivelsesdato: 1978-null...

  15. Neutrophils, lymphocytes, and monocytes exhibit diverse behaviors in transendothelial and subendothelial migrations under coculture with smooth muscle cells in disturbed flow.

    Science.gov (United States)

    Chen, Cheng-Nan; Chang, Shun-Fu; Lee, Pei-Ling; Chang, Kyle; Chen, Li-Jing; Usami, Shunichi; Chien, Shu; Chiu, Jeng-Jiann

    2006-03-01

    Atherosclerosis develops at regions of the arterial tree exposed to disturbed flow. The early stage of atherogenesis involves the adhesion of leukocytes (white blood cells [WBCs]) to and their transmigration across endothelial cells (ECs), which are located in close proximity to smooth muscle cells (SMCs). We investigated the effects of EC/SMC coculture and disturbed flow on the adhesion and transmigration of 3 types of WBCs (neutrophils, peripheral blood lymphocytes [PBLs], and monocytes) using our vertical-step flow (VSF) chamber, in which ECs were cocultured with SMCs in collagen gels. Such coculture significantly increased the adhesion and transmigration of neutrophils, PBLs, and monocytes under VSF, particularly in the reattachment area, where the rolling velocity of WBCs and their transmigration time were decreased, as compared with the other areas. Neutrophils, PBLs, and monocytes showed different subendothelial migration patterns under VSF. Their movements were more random and shorter in distance in the reattachment area. Coculture of ECs and SMCs induced their expressions of adhesion molecules and chemokines, which contributed to the increased WBC adhesion and transmigration. Our findings provide insights into the mechanisms of WBC interaction with the vessel wall (composed of ECs and SMCs) under the complex flow environments found in regions of prevalence for atherogenesis.

  16. Flow cytometry of cerebrospinal fluid (CSF) lymphocytes: alterations of blood/CSF ratios of lymphocyte subsets in inflammation disorders of human central nervous system (CNS).

    Science.gov (United States)

    Kleine, T O; Albrecht, J; Zöfel, P

    1999-03-01

    Flow cytometry was adapted to measure lymphocytes in human cerebrospinal fluid (CSF). The method was sufficiently precise, reproducible and accurate despite low cell counts. In lumbar CSF of controls with 500 to 3500 (10(3)/l) leukocytes, lymphocyte counts correlated with those in corresponding venous blood: blood/CSF ratios of approximately 2000 : 1 were found for total T cells (CD3+) and CD3+ HLA-DR-, CD3+4+, CD3+8+ subsets, ratios were increased for the lymphocyte subsets CD3+ HLA-DR+ blood-brain and blood-CSF barriers) to blood lymphocyte subsets which favor the transfer of T subsets. Correlation of the subset ratios to the CD3+ ratio indicates distinct barrier properties which changed differently with acute and subacute inflammations and neuroimmunological diseases of central nervous system (CNS) in lumbar or ventricular CSF, but not with simple protein barrier disturbance. HLA DR+ T ratios were higher than HLA DR- T ratios only with controls and some neuroimmunological diseases. Lymphocyte barrier characteristics were related to protein leakage situated at the same barriers, indicating for the lymphocyte subsets selective transfer routes in control subjects and non-selective routes in patients with CNS inflammation where altered ratios revealed a mixture of both routes.

  17. [Production of interleukin-2 by peripheral blood lymphocytes from patients with soft tissue sarcomas].

    Science.gov (United States)

    Berezhnaia, N M; Goretskiĭ, B A; Konovalenko, V F; Palivets, A Iu; Tolstopiatov, B A

    1987-01-01

    Interleukin-2 (IL-2) production of phytohemagglutinin-stimulated peripheral blood lymphocytes (PBL) was studied in 9 healthy subjects and 19 patients with soft tissue sarcomas. Mean IL-2 production by PBL in 19 patients was significantly diminished as compared with the control. Surgery leads to an increase of IL-2 production, however, the levels observed in the control do not restore completely.

  18. The Assessment of Cytotoxicity and Genotoxicity of Mirtazapine in Human Blood Lymphocytes Using Micronucleus Test

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    M Norizadeh tazehkand

    2015-02-01

    Results: MN formation was not significantly induced at 24- and 48-h treatment periods when compared with control but Nuclear division index (NDI significantly decreased at all concentrations for two treatment periods. Conclusion: Mirtazapine was not genetoxic but was cytotoxic in human peripheral blood lymphocytes. According to this study mirtazapine has cytotoxic effects on human's cells.

  19. LYMPHOCYTE SUBSETS AND CYTOKINES IN BLOOD AND CEREBROSPINAL FLUID IN CHILDREN WITH VIRAL AND BACTERIAL MENINGITIS

    Directory of Open Access Journals (Sweden)

    L. A. Alekseeva

    2016-01-01

    Full Text Available Introduction of flow cytometry caused an increase in the investigation of liquor lymphocyte pool phenotype in the case of different brain disorders, including viral and bacterial meningitis, however this type of research in children has been relatively rare. Phenotype and lymphocyte functions are under cytokine control system, therefore detection of interconnections between lymphocyte pool subpopulation composition and cytokine level in blood and liquor of the patients concerns a great interest. The purpose of this research was to study lymphocyte subpopulation composition and the level of cytokines IL-1β, IL-6, IL-8, IL-10, IFNα, IFNγ and IL-4, and also IgG in liquor and blood of children with viral and bacterial meningitis. There was performed blood and liquor investigation in 46 children aged from 1 to 16 years old with viral (n = 35 and bacterial (n = 11 meningitis. Immunophenotyping of blood and liquor cells was performed by the method of flow cytometry with the use of monoclonal antibodies to CD3, CD4, CD8, CD19, CD16, CD56, CD25 and CD95. The content of cytokines was detected in ELISA, and that of IgG — by the method of quantitative immunoturbodimetry. During an acute period of viral meningitis there was detected a decrease in NK portion and activated CD25+ cells in the blood of patients accompanied by the increase in B-lymphocytes number, along with cytokine IFNγ, IL-8 and IL-10 serum level rise. There was determined T-lymphocytes accumulation in liquor with the prevalence of CD4+ Т-cells and, to a lesser degree, CD25+ and CD95+ cells, NK and B-lymphocytes. Intrathecally there was noted the predominance of IL-6 response accompanied by the growth of IL-8 and IL-10 concentration as well. During an acute period of bacterial meningitis there was noted a decrease in percentage of CD3+, CD4+, CD8+ Т-lymphocytes, NK, CD25+ and CD95+ cells, along with, on the contrary, sharp increase in B-cells pool, simultaneously with

  20. Leukocytic Response and Peripheral Venous Blood Lymphocyte Apoptosis as a Marker of Tissue Ischemia in Acute Massive Blood Loss

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    N. V. Borovkova

    2013-01-01

    Full Text Available Objective: to estimate the level of peripheral venous blood lymphocyte apoptosis and intraoperative hypoxia in victims with acute massive blood loss. Subjects and methods. Twenty-two patients with open and close chest and abdominal traumas complicated by acute massive blood loss were examined. All the patients were emergently operated on to stop bleeding. Tissue metabolism was evaluated from gases, acid-base parameters, and plasma lactate, glucose, potassium, and sodium levels. Apoptosis of mononuclear cells was studied and dead leukocytes were counted using flow cytometry. Results. Preoperatively, the victims were found to have venous hypoxemia, hyperlactatemia, hyperglycemia, moderate leukocytosis, and higher dead leukocyte counts. There were also raised counts of lymphocytes coming into the process of apoptosis. A significant relationship was found between monocyte counts and hypoxia values. At the end of surgery, oxygen balance values became stable and exerted an effect on the count of leukocytes, the relative level of granulocytes, the relative and absolute counts of dead and damaged leukocytes, and the concentration of lymphocytes in the victims’ venous blood during the early stages of apoptosis, as evidenced by nonlinear regression models. Conclusion. The indicators of immunocompetent cell apoptosis and the count of venous blood dead leukocytes along with lactate levels and venous hypoxemia parameters reflect the degree of tissue hypoxia and may be used as specific markers.

  1. Protective effect of quercetin against oxidative stress caused by dimethoate in human peripheral blood lymphocytes

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    Lassoued Saloua

    2011-08-01

    Full Text Available Abstract Background The aim of this study is to investigate the effect of quercetin in alleviating the cytotoxic effects of Dimethoate in human peripheral blood lymphocytes. Methods Lymphocytes were divided into too groups. The first group, lymphocytes were incubated for 4 h at 37°C with different concentrations (0, 40, 60 and 100 mM of Dimethoate. The second group was preincubated with quercetin for 30 min and followed by Dim incubation for 4 h at 37°C. Results Following in vitro incubation, Dimethoate caused a significant increase in malondialdehyde levels, a significant decrease in thiol levels, as well as a significant increase in superoxide dismutase, and catalase activities in lymphocytes at different concentrations. Quercetin pretreated lymphocytes showed a significant protection against the cytotoxic effects inducted by Dimethoate on the studied parameters. Conclusion In conclusion, antioxidant quercetin could protect against Dimethoate-induced oxidative stress by decreasing lipid peroxidation, protein oxidation and increasing superoxide dismutase and catalase activities in human lymphocytes.

  2. Type and Portions of Peripheral Blood T Lymphocytes in Oral Lichen Planus

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    Kia

    2013-06-01

    Full Text Available Background Lichen planus is a disease with unknown etiology that affects the skin and the mucous membranes. Immune dysregulation in the pathogenesis of oral lichen planus (OLP is well-known phenomenon. Objectives In this study, we compared the levels of the peripheral blood T lymphocytes between patients with OLP and control group. Patients and Methods In this study, 32 and 16 patients respectively with and without OLP were recruited. Five milliliters of the participants' peripheral venous blood was drew in an EDTA-containing test tube and the levels of CD3+, CD4+, and CD8+ cells, CD4+/CD8+ and CD4+/CD3+ ratio were measured by means of two-color flow cytometry. The data were analyzed in SPPS v.19 by employing Mann-Whitney U test. Results There were no significant difference among the levels of CD3+, CD4+, and CD8+ lymphocytes and the ratio of CD4+/CD8+ and CD4+/CD3+ lymphocytes between patients and control group; however, there was a significant difference between male and female patients with respect to the levels of CD3+ and CD4+ lymphocytes and the ratio of CD4+/CD8+ and CD4+/CD3+ lymphocytes. Conclusions Our results confirm that only local immune mechanism known as skin-associated lymphoid tissue, not a systemic immunologic disorder, was involved in the OLP.

  3. Apoptotic cell death, detected ex vivo in peripheral blood lymphocytes of HIV-1 infected persons

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    L. F. te Velde

    1996-01-01

    Full Text Available In HIV-1 infection the ongoing depletion of CD4+ T-lymphocytes is believed, to a large extent, to be due to apoptosis. Until now quantitative information about in vivo apoptosis of lymphocytes in HIV-patients is scarce because of the very nature of the apoptotic process. Successful detection of apoptosis ex vivo requires the recognition of the initial phase of this process, because at a later stage the cells may not remain any longer in the circulation. We measured quantitatively the amount of early apoptotic peripheral blood lymphocytes directly ex vivo in HIV-1 infected patients using a recently described flow cytometric assay. With this method we observed in an unselected heterogenous group of twelve HIV-infected individuals a median percentage of apoptotic lymphocytes to be significantly higher than in ten healthy controls. To the best of our knowledge this is the first report of ex vivo observed increased apoptosis of peripheral blood lymphocytes in HIV-infected persons.

  4. Changes in hematological indices and lymphocyte subsets in response to whole blood donation in healthy male donors.

    Science.gov (United States)

    Borai, Anwar; Livingstone, Callum; Alsobhi, Enaam; Al Sofyani, Abeer; Balgoon, Dalal; Farzal, Anwar; Almohammadi, Mohammed; Al-Amri, Abdulafattah; Bahijri, Suhad; Alrowaili, Daad; Bassiuni, Wafaa; Saleh, Ayman; Alrowaili, Norah; Abdelaal, Mohamed

    2017-04-01

    Whole blood donation has immunomodulatory effects, and most of these have been observed at short intervals following blood donation. This study aimed to investigate the impact of whole blood donation on lymphocyte subsets over a typical inter-donation interval. Healthy male subjects were recruited to study changes in complete blood count (CBC) (n = 42) and lymphocyte subsets (n = 16) before and at four intervals up to 106 days following blood donation. Repeated measures ANOVA were used to compare quantitative variables between different visits. Following blood donation, changes in CBC and erythropoietin were as expected. The neutrophil count increased by 11.3% at 8 days (p donation, there are transient changes in lymphocyte subsets. The effect of BMI on lymphocyte subsets and the effect of this immunomodulation on the immune response merit further investigation.

  5. Intracerebroventricular infusions of TNF-alpha preferentially recruit blood lymphocytes and induce a perivascular leukocyte infiltrate.

    Science.gov (United States)

    Seabrook, T J; Hay, J B

    2001-02-01

    Tumour necrosis factor (TNF)-alpha is important in several central nervous system (CNS) inflammatory diseases, however, its role in the recruitment of leukocytes into the cerebral spinal fluid (CSF) and CNS is incompletely understood. Therefore, we examined the effect of intracerebroventricular (icv) and parenchymal infusions of TNF-alpha on the type of leukocyte, the pool and subset of lymphocytes recruited into CSF and brain parenchyma. Parenchymal injections of 500 ng of recombinant human TNF-alpha did not induce inflammation, whereas an icv infusion of TNF-alpha caused CSF leuckocytosis and a perivascular infiltrate. Twenty-four hours after the icv infusion neutrophils predominated, with CD4+ T cells being the major lymphocyte subset in CSF. By 48 h lymphocytes were the dominant cell type with CD8+ cells surpassing CD4+ cells in both the CSF and the perivascular infiltrate. The labeled recirculating lymphocyte pool prevailed in normal CSF, but after the infusion of TNF-alpha, the blood pool of lymphocytes was preferentially recruited. These results have implications for the immune surveillance of the CNS.

  6. REPRESENTATION OF DIFFERENT LYMPHOCYTES' POPULATIONS IN PERIPHERAL BLOOD OF PATIENTS WITH UTERINE MYOMA

    Directory of Open Access Journals (Sweden)

    Ye. E. Zueva

    2014-07-01

    Full Text Available Abstract. Uterine myoma is one of the most widespread gynecological pathology among reproductive women older than 30 years. It is known, that often progress of this pathology is associated with genetic and endocrinologic factors. The immune system is not evident still. The aim of this study was to analyze the state of patient's immune system using flow cytometry assessment of different subpopulations of lymphocytes in peripheral blood. We have examined 46 patients with simple and proliferating forms of the myoma, with different variants of clinical symptoms. Absolute and relative content of different subpopulations of lymphocytes was not differed from normal population's standard. Significant differences of B-lymphocytes and natural killers content were observed between groups with simple and proliferating forms of disease. It was shown that metrorrhagia is associated with high level of T-lymphocytes and T-killers. It was noted that decreasing of B-lymphocytes content took place in cases with large number of uterine nodes. Obtained data are not sufficient for complete understanding of the role of immune system in pathogenesis of this disease, but they confirm that using of immunomodulating therapy is expedient for complex treatment of uterine myoma.

  7. In vitro cytotoxic, genotoxic and antioxidant/oxidant effects of guaiazulene on human lymphocytes

    Directory of Open Access Journals (Sweden)

    Başak Toğar

    2015-02-01

    Full Text Available The aim of this study was to evaluate for the cytotoxicity, genotoxicity and antioxidant/oxidant activity of GYZ on human peripheral blood lymphocytes (PBLs. Guaiazulene (GYZ was added into culture tubes at various concentrations (0-400 µg/mL-1. Cytotoxicity against the human lymphocytes cultures was examined by lactate dehydrogenase (LDH release assay. The proliferative response was estimated by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT assay. Antioxidant/oxidant activity was evaluated by measuring the total oxidant status (TOS and total antioxidant capacity (TAC levels. Micronucleus (MN and chromosomal aberration (CA tests were used in genotoxicity studies. The results showed that GYZ caused cytotoxicity in the PBLs at high concentrations, but TOS level were not affected, while the level of TAC was significantly increased. GYZ also did not induce chromosomal aberrations when compared to that of the control group. Results this study clearly revealed that GYZ was not genotoxic and also increased the capacity of the antioxidant in the culture of human PBL cells. This report is first report on the impact of GYZ on human PBL cells.

  8. No evidence of transmission of chronic lymphocytic leukemia through blood transfusion

    DEFF Research Database (Denmark)

    Hjalgrim, Henrik; Rostgaard, Klaus; Vasan, Senthil K;

    2015-01-01

    Monoclonal B-cell lymphocytosis (MBL) is a precursor of chronic lymphocytic leukemia (CLL). Observations of MBL in blood donors raise concern that transmitted MBL may cause recipient CLL. Using a database with health information on 1.5 million donors and 2.1 million recipients, we compared CLL oc...... transfusion experience over more than 30 years indicate that MBL/CLL transmission does not contribute importantly to recipient CLL risk....

  9. Expression of membrane receptor for tumour necrosis factor on human blood lymphocytes.

    Science.gov (United States)

    Zola, H; Flego, L; Weedon, H

    1993-08-01

    Using a monoclonal antibody against the human p75 tumour necrosis factor receptor (TNFR-I) combined with a high-sensitivity immunofluorescence flow cytometric procedure, a proportion of peripheral blood lymphocytes can be shown to express TNFR-I constitutively. Approximately 50% of peripheral blood lymphocytes consisting mostly of CD4 cells and including most CD45R0-positive cells, express TNFR-I. Receptor expression is increased by a variety of activation signals. Only a minority (up to 30%) of tonsil B cells express measurable levels of TNFR-I. The tonsil B cells which express TNFR-I include both cells with a germinal centre cell phenotype and cells with the phenotype of the follicular mantle zone. Activation of B cells with anti-immunoglobulin, alone or in combination with interleukin-4 or interleukin-2, increases receptor expression, particularly in cells with the phenotype of mantle zone cells. The functional significance of constitutive expression of TNFR by blood and tissue lymphocytes is discussed.

  10. Fas and Bcl-2 Expression on T Lymphocyte Subsets in the Peripheral Blood of Relapsing Patients with Condyloma Acuminatum

    Institute of Scientific and Technical Information of China (English)

    顾军; 范清源; 高春芳; 代夫; 郑茂荣

    2003-01-01

    Objective: To study the expression of Fas and Bcl-2 proteins on T lymphocyte subsets in the peripheral blood of relapsing patients with condyloma acuminatum(CA) and healthy controls.Methods: Flow cytometry (permeabization and staining procedure with conjugated antibodies) was used.Results: We observed that the expression of Fas protein on CD4+ T lymphocyte subset of CA patients was significantly higher than that of healthy controls( P<0.01 ).Conclusions: Increased expression of Fas proteinon CD4+ T lymphocyte subset may be a cause of de-creased percentage of CD4+ T lymphocyte subset. This induces the increased ratio of CD4+/CD8+.

  11. Effects of Immunopotentiator of the Traditional Chinese Medicine on T Lymphocytes in Chicken Blood

    Institute of Scientific and Technical Information of China (English)

    SHI Qiumei; LI Chunling; GAO Guisheng; SHEN Ping; TANG Shengling

    2008-01-01

    In order to investigate the mechanism of action of immunoenhancer, the effects of the traditional Chinese medicine immunopromoter on the quantity and the transformation rates of T lymphocytes in the chicken blood were determined. Total 120 chickens were randomly assigned into three groups. The 1% and the 0.5% of the Chinese medicine immunopromoter were added to the chicken drinking water, respectively. The quantity of T lymphocytes in each group was measured by a-Naphthyl acetate esterase (ANAE) staining. The results showed that the percentages of T lymphocytes of the treatment groups were significantly higher than that of the control group (P<0.05), and the percentage of the 1% group significantly higher than that of the 0.5% group (P<0.05). In conclusion, the transformation rates of T lymphocytes showed that the Chinese medicine immunopromoter had the significant enhancing effect on the transformation rates of T lymphoeytes of the treated chickens. The traditional Chinese medicine immunopromoter had the distinct function to promote the quantity and the transformation rate of T iymphocytes.

  12. DNA damage in blood lymphocytes in patients after {sup 177}Lu peptide receptor radionuclide therapy

    Energy Technology Data Exchange (ETDEWEB)

    Eberlein, Uta; Bluemel, Christina; Buck, Andreas Konrad; Werner, Rudolf Alexander; Lassmann, Michael [University of Wuerzburg, Department of Nuclear Medicine, Wuerzburg (Germany); Nowak, Carina; Scherthan, Harry [Bundeswehr Institute of Radiobiology affiliated to the University of Ulm, Munich (Germany)

    2015-10-15

    The aim of the study was to investigate DNA double strand break (DSB) formation and its correlation with the absorbed dose to the blood lymphocytes of patients undergoing their first peptide receptor radionuclide therapy (PRRT) with {sup 177}Lu-labelled DOTATATE/DOTATOC. The study group comprised 16 patients receiving their first PRRT. At least six peripheral blood samples were obtained before, and between 0.5 h and 48 h after radionuclide administration. From the time-activity curves of the blood and the whole body, residence times for blood self-irradiation and whole-body irradiation were determined. Peripheral blood lymphocytes were isolated, fixed with ethanol and subjected to immunofluorescence staining for colocalizing γ-H2AX/53BP1 DSB-marking foci. The average number of DSB foci per cell per patient sample was determined as a function of the absorbed dose to the blood and compared with an in vitro calibration curve established in our laboratory with {sup 131}I and {sup 177}Lu. The average number of radiation-induced foci (RIF) per cell increased over the first 5 h after radionuclide administration and decreased thereafter. A linear fit from 0 to 5 h as a function of the absorbed dose to the blood agreed with our in vitro calibration curve. At later time-points the number of RIF decreased, indicating progression of DNA repair. Measurements of RIF and the absorbed dose to the blood after systemic administration of {sup 177}Lu may be used to obtain data on the individual dose-response relationships in vivo. Individual patient data were characterized by a linear dose-dependent increase and an exponential decay function describing repair. (orig.)

  13. Fludarabine Phosphate and Total-Body Irradiation Before Donor Peripheral Blood Stem Cell Transplant in Treating Patients With Chronic Lymphocytic Leukemia or Small Lymphocytic Leukemia

    Science.gov (United States)

    2016-07-18

    B-Cell Prolymphocytic Leukemia; Chronic Lymphocytic Leukemia; Prolymphocytic Leukemia; Recurrent Chronic Lymphocytic Leukemia; Recurrent Small Lymphocytic Lymphoma; Refractory Chronic Lymphocytic Leukemia; T-Cell Prolymphocytic Leukemia

  14. Peripheral blood lymphocyte HIV DNA levels correlate with HIV associated neurocognitive disorders in Nigeria.

    Science.gov (United States)

    Jumare, Jibreel; Sunshine, Sara; Ahmed, Hayat; El-Kamary, Samer S; Magder, Laurence; Hungerford, Laura; Burdo, Tricia; Eyzaguirre, Lindsay M; Umlauf, Anya; Cherner, Mariana; Abimiku, Alash'le; Charurat, Man; Li, Jonathan Z; Blattner, William A; Royal, Walter

    2017-02-27

    Mononuclear cells play key roles in the pathogenic mechanisms leading to HIV-associated neurocognitive disorders (HANDs). We examined the association between HIV DNA within peripheral blood mononuclear cell (PBMC) subsets and HAND in Nigeria. PBMCs were collected at baseline from 36 antiretroviral naive participants. CD14+ cells and T&B lymphocyte fractions were isolated by, respectively, positive and negative magnetic bead separation. Total HIV DNA within CD14+ and T&B cells were separately quantified using real-time PCR assay targeting HIV LTR-gag and cell input numbers determined by CCR5 copies/sample. Utilizing demographically adjusted T scores obtained from a 7-domain neuropsychological test battery, cognitive status was determined by the global deficit score (GDS) approach, with a GDS of ≥0.5 indicating cognitive impairment. In a linear regression adjusting for plasma HIV RNA, CD4 and lymphocyte count, Beck's depression score, and years of education, there was 0.04 lower log10 HIV DNA copies within T&B lymphocytes per unit increase in global T score (p = 0.02). Adjusting for the same variables in a logistic regression, the odds of cognitive impairment were 6.2 times greater per log10 increase in HIV DNA within T&B lymphocytes (p = 0.048). The association between cognitive impairment and HIV DNA within CD14+ monocytes did not reach statistical significance. In this pretreatment cohort with mild cognitive dysfunction, we found a strong association between levels of HIV DNA within the lymphocyte subset and HAND independent of plasma HIV RNA. These findings likely reflect the neurologic impact of a larger HIV reservoir and active viral replication.

  15. Cytogenetic studies of blood lymphocytes from cosmonauts after long-term space flights on MIR station

    Science.gov (United States)

    Fedorenko, B.; Druzhinin, S.; Yudaeva, L.; Petrov, V.; Akatov, Yu.; Snigiryova, G.; Novitskaya, N.; Shevchenko, V.; Rubanovich, A.

    Long-term space missions may increase risks of unfavorable consequences for cosmonauts as a result of radiation effects. This paper presents results of a study of cytogenetic damage in cosmonauts' peripheral blood lymphocytes induced by space radiation. Cultivation of lymphocytes and analysis of chromosomal aberrations were made according to generally accepted methods. It is shown that the yields of dicentrics and centric rings scored after long-term space flights are considerably higher than those scored prior to the flights. An attempt was made to assess individual doses received by cosmonauts. Individual biodosimetry doses received by cosmonauts who showed a reliable increase in the yields of chromosomal-type aberrations after their first flights were estimated to be from 0.02 to 0.28 Gy.

  16. [Rapid dicentric assay of human blood lymphocytes after exposure to low doses of ionizing radiation].

    Science.gov (United States)

    Repin, M V; Repina, L A

    2011-01-01

    The probability of losses of different chromosome aberrations during the dicentric chromosome assay of metaphase cells with incomplete sets of chromosome centromeres was estimated using a mathematical model for low doses of ionizing radiation. A dicentric assay of human blood lymphocytes without determination of the total amount of chromosome centromeres in cells without chromosome aberrations (rapid dicentric assay) has been proposed. The rapid dicentric analysis allows to register chromosome aberrations in full compliance with the conventional classification. The experimental data have shown no statistically significant difference between the frequencies of dicentric chromosomes detected by rapid and classical dicentric chromosome assays of human lymphocytes exposed to 0.5 Gy of 60Co gamma-rays. The rate of the rapid dicentric assay was almost twice as high as that of the classical dicentric assay.

  17. DNA strand breaks (comet assay) in blood lymphocytes from wild bottlenose dolphins.

    Science.gov (United States)

    Lee, Richard F; Bulski, Karrie; Adams, Jeffrey D; Peden-Adams, Margie; Bossart, Gregory D; King, Lydia; Fair, Patricia A

    2013-12-15

    The comet assay was carried out on blood lymphocytes from a large number of wild dolphins (71 from Indian River Lagoon, FL, USA; 51 from Charleston Harbor, SC, USA) and provides a baseline study of DNA strand breaks in wild dolphin populations. There were no significant differences in the comet assay (% DNA in tail) results between the different age and sex categories. Significant difference in DNA strand breaks were found between Charleston Harbor dolphins (median--17.4% DNA in tail) and Indian River Lagoon dolphins (median--14.0% DNA in tail). A strong correlation found between T-cell proliferation and DNA strand breaks in dolphin lymphocytes suggests that dolphins with a high numbers of DNA strand breaks have a decreased ability to respond to infection. Higher concentrations of genotoxic agents in Charleston Harbor compared with Indian River lagoon may have been one of the causes of higher DNA strand breaks in these dolphins.

  18. Blood mean platelet volume and platelet lymphocyte ratio as new predictors of hip osteoarthritis severity.

    Science.gov (United States)

    Taşoğlu, Özlem; Şahin, Ali; Karataş, Gülşah; Koyuncu, Engin; Taşoğlu, İrfan; Tecimel, Osman; Özgirgin, Neşe

    2017-02-01

    Osteoarthritis (OA) is a low grade systemic inflammatory disease in which many inflammatory mediators are known to be elevated in the peripheric blood. Blood platelet lymphocyte ratio (PLR) and mean platelet volume (MPV) are accepted as novel markers in many of the systemic inflammatory disorders, but have not been investigated in synovitis-free radiographic OA yet.The aim of this study was to evaluate the levels of blood PLR and MPV in radiographic hip OA. A total of 880 patients were evaluated retrospectively and after certain exclusion criteria, 237 of them who have primary hip OA were included. Age, sex, height, weight, body mass index, neutrophil, lymphocyte and platelet counts, erythrocyte sedimentation rate (ESR), PLR, and MPV levels were recorded, Kellgren-Lawrence (KL) grading of the hip joints were performed. Patients were then divided into 2 groups as KL grades 1 to 2 (mild-moderate) and KL grades 3 to 4 (severe) hip OA.Mean age, mean neutrophil, lymphocyte and platelet counts, mean MPV, mean PLR, and mean ESR were statistically significantly different between mild/moderate hip OA group and severe hip OA group. In univariate analysis, older age and higher MPV, PLR, and ESR were severely associated with severe hip OA. In multiple logistic regression analysis, MPV, PLR, and ESR emerged as independent predictors of severe hip OA.The results of the present study, for the first time in the literature, suggest blood PLR and MPV as novel inflammatory markers predicting the radiographic severity of hip OA in the daily practice.

  19. Impaired NADPH oxidase activity in peripheral blood lymphocytes of galactosemia patients.

    Science.gov (United States)

    Al-Essa, Mazen; Dhaunsi, Gursev S; Al-Qabandi, Wafa'a; Khan, Islam

    2013-07-01

    Galactosemia is an autosomal recessive disorder with a wide range of clinical abnormalities. Cellular oxidative stress is considered as one of the pathogenic mechanisms of galactosemia. In this study, we examined the activity of NADPH oxidase (NOX), a major superoxide-generating enzyme system, in peripheral blood lymphocytes (PBL) from galactosemia patients. PBL were isolated from galactosemia patients and healthy control subjects and used for cell culture studies and biochemical assays. PBL were cultured in the presence or absence of galactose or galactose-1-phosphate (Gal-1-P), and enzyme activities and/or gene expression of NOX, catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPx) were measured in the cell homogenates. PBL isolated from galactosemia patients showed significantly reduced (P Galactosemia patients were found to have significantly (P galactosemia patients; however, Western blotting revealed that NOX-1 protein was not significantly altered. Interestingly, levels of NOX activity in lymphocytes isolated from galactosemia patients significantly increased but remained subnormal when cultured in galactose-deficient medium for two weeks, indicating a galactose-mediated inhibition of NOX. Lymphocytes isolated from control subjects were found to have significantly (P galactosemia patients.

  20. Peripheral blood lymphocyte subsets in Fasciola hepatica infected and immunised goats.

    Science.gov (United States)

    Zafra, R; Pérez, J; Buffoni, L; Martínez-Moreno, F J; Acosta, I; Mozos, E; Martínez-Moreno, A

    2013-09-01

    The proportions of CD4(+), CD8(+) and WC1+ T lymphocytes from peripheral blood using flow cytometry were investigated in goats infected with Fasciola hepatica and previously immunised with recombinant Cathepsin-L1 (rCL1) and Glutathione-S-transferase sigma class (GST). The immunisation trial did not induce protective responses, and no significant differences were recorded between immunised and non-immunised groups. However, there was a significant decrease in the proportion of CD4(+) T lymphocytes in the infected groups both at 5 weeks post-infection (wpi), coinciding with the migratory stage of the infection, and at 12 wpi in the biliary stage of the infection. The proportional decrease in this circulating population may be related to the recruitment of CD4(+) T cells in liver and hepatic lymph nodes and also to the immunomodulatory effect of the parasite through the interaction of F. hepatica excretory-secretory products (FhESP) with this cell population. To date, this is the first report about the effect of F. hepatica infection in peripheral lymphocyte subsets in goats.

  1. Expression of blood serum proteins and lymphocyte differentiation clusters after chronic occupational exposure to ionizing radiation.

    Science.gov (United States)

    Rybkina, Valentina L; Azizova, Tamara V; Scherthan, Harry; Meineke, Viktor; Doerr, Harald; Adamova, Galina V; Teplyakova, Olga V; Osovets, Sergey V; Bannikova, Maria V; Zurochka, Alexander V

    2014-11-01

    This study aimed to assess effects of chronic occupational exposure on immune status in Mayak workers chronically exposed to ionizing radiation (IR). The study cohort consists of 77 workers occupationally exposed to external gamma-rays at total dose from 0.5 to 3.0 Gy (14 individuals) and workers with combined exposure (external gamma-rays at total dose range 0.7-5.1 Gy and internal alpha-radiation from incorporated plutonium with a body burden of 0.3-16.4 kBq). The control group consists of 43 age- and sex-matched individuals who never were exposed to IR, never involved in any cleanup operations following radiation accidents and never resided at contaminated areas. Enzyme-linked immunoassay and flow cytometry were used to determine the relative concentration of lymphocytes and proteins. The concentrations of T-lymphocytes, interleukin-8 and immunoglobulins G were decreased in external gamma-exposed workers relative to control. Relative concentrations of NKT-lymphocytes, concentrations of transforming growth factor-β, interferon gamma, immunoglobulins A, immunoglobulins M and matrix proteinase-9 were higher in this group as compared with control. Relative concentrations of T-lymphocytes and concentration of interleukin-8 were decreased, while both the relative and absolute concentration of natural killers, concentration of immunoglobulins A and M and matrix proteinase-9 were increased in workers with combined exposure as compared to control. An inverse linear relation was revealed between absolute concentration of T-lymphocytes, relative and absolute concentration of T-helpers cells, concentration of interferon gamma and total absorbed dose from external gamma-rays in exposed workers. For workers with incorporated plutonium, there was an inverse linear relation of absolute concentration of T-helpers as well as direct linear relation of relative concentration of NKT-lymphocytes to total absorbed red bone marrow dose from internal alpha-radiation. In all, chronic

  2. Effects of cerebrolysin administration on oxidative stress-induced apoptosis in lymphocytes from CADASIL patients.

    Science.gov (United States)

    Formichi, Patrizia; Radi, Elena; Battisti, Carla; Di Maio, Giuseppe; Dotti, Maria Teresa; Muresanu, Dafin; Federico, Antonio

    2013-04-01

    Cerebrolysin (Cere) is a peptidergic nootropic drug with neurotrophic properties which has been used to treat dementia and sequelae of stroke. Use of Cere prevents nuclear structural changes typical of apoptosis and significantly reduces the number of apoptotic cells after several apoptotic stimuli. Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) is a hereditary disease caused by mutations of the Notch3 gene encoding the Notch3 protein. Notch3 is involved in the regulation of apoptosis, modulating Fas-Ligand (Fas-L)- induced apoptosis. The aim of this study was to evaluate the in vitro protective effects of Cere against oxidative stress-induced apoptosis in cells from CADASIL patients. We used peripheral blood lymphocytes (PBLs) from 15 CADASIL patients (age range 34-70 years); 2-deoxy-D-ribose (dRib), a highly reducing sugar, was used as paradigm pro-apoptotic stimulus. Apoptosis was analyzed by flow cytometry and fluorescence microscopy. Administration of Cere to PBLs from CADASIL patients cultured under standard conditions had no effect on the percentage of apoptotic cells. Administration of Cere to PBLs cultured with dRib caused a significant decrease in apoptosis after 48 h of culture in only 5 patients, whereas in the other 10 patients, Cere treatment was not associated with any significant difference in the percentage of apoptosis. This result showed a protective effect of Cere against oxidative stress-induced apoptosis only in 30 % of the CADASIL patients, suggesting that the Notch3 gene probably does not influence the anti-apoptotic properties of Cere in vitro.

  3. Human immune compartment comparisons: Optimization of proliferative assays for blood and gut T lymphocytes.

    Science.gov (United States)

    Dock, Jeffrey; Hultin, Lance; Hultin, Patricia; Elliot, Julie; Yang, Otto O; Anton, Peter A; Jamieson, Beth D; Effros, Rita B

    2017-03-21

    The accumulation of peripheral blood late-differentiated memory CD8 T cells with features of replicative (cellular) senescence, including inability to proliferate in vitro, has been extensively studied. Importantly, the abundance of these cells is directly correlated with increased morbidity and mortality in older persons. Of note, peripheral blood contains only 2% of the total body lymphocyte population. By contrast, the gut-associated lymphoid tissue (GALT) is the most extensive lymphoid organ, housing up to 60% of total body lymphocytes, but has never been assessed with respect to senescence profiles. We report here the development of a method for measuring and comparing proliferative capacity of peripheral blood and gut colorectal mucosa-derived CD8 T cells. The protocol involves a 5-day culture of mononuclear leukocyte populations, from blood and gut colorectal mucosa respectively, labeled with 5-(and 6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) and 5-bromo-2'-deoxyuridine (BrdU) and stimulated with anti-CD2/3/28-linked microbeads. Variables tested and optimized as part of the protocol development include: mode of T cell stimulation, CFSE concentration, inclusion of a second proliferation marker, BrdU, culture duration, initial culture concentration, and inclusion of autologous irradiated feeder cells. Moving forward, this protocol demonstrates a significant advance in the ability of researchers to study compartment-specific differences of in vitro proliferative dynamics of CD8 T cells, as an indicator of replicative senescence and immunological aging. The study's two main novel contributions are (1) Optimization and adaptation of standard proliferative dynamics blood T cell protocols for T cells within the mucosal immune system. (2) Introduction of the novel technique of combining CFSE and BrdU staining to do so.

  4. Infants' Peripheral Blood Lymphocyte Composition Reflects Both Maternal and Post-Natal Infection with Plasmodium falciparum.

    Directory of Open Access Journals (Sweden)

    Odilon Nouatin

    Full Text Available Maternal parasitoses modulate fetal immune development, manifesting as altered cellular immunological activity in cord blood that may be linked to enhanced susceptibility to infections in early life. Plasmodium falciparum typifies such infections, with distinct placental infection-related changes in cord blood exemplified by expanded populations of parasite antigen-specific regulatory T cells. Here we addressed whether such early-onset cellular immunological alterations persist through infancy. Specifically, in order to assess the potential impacts of P. falciparum infections either during pregnancy or during infancy, we quantified lymphocyte subsets in cord blood and in infants' peripheral blood during the first year of life. The principal age-related changes observed, independent of infection status, concerned decreases in the frequencies of CD4+, NKdim and NKT cells, whilst CD8+, Treg and Teff cells' frequencies increased from birth to 12 months of age. P. falciparum infections present at delivery, but not those earlier in gestation, were associated with increased frequencies of Treg and CD8+ T cells but fewer CD4+ and NKT cells during infancy, thus accentuating the observed age-related patterns. Overall, P. falciparum infections arising during infancy were associated with a reversal of the trends associated with maternal infection i.e. with more CD4+ cells, with fewer Treg and CD8+ cells. We conclude that maternal P. falciparum infection at delivery has significant and, in some cases, year-long effects on the composition of infants' peripheral blood lymphocyte populations. Those effects are superimposed on separate and independent age- as well as infant infection-related alterations that, respectively, either match or run counter to them.

  5. Age-related changes of the multidrug resistance P-glycoprotein function in normal human peripheral blood T lymphocytes

    Directory of Open Access Journals (Sweden)

    C.G. Machado

    2003-12-01

    Full Text Available The multidrug resistance P-glycoprotein is a transmembrane efflux pump expressed by lymphocytes and is involved in their cytolytic activity. In the present study, we investigated the age-related changes of P-glycoprotein function in normal peripheral blood lymphocytes. Blood samples from 90 normal volunteers (age range, 0 to 86 years were analyzed. P-glycoprotein function was assessed by the flow cytometric rhodamine 123 assay. P-glycoprotein function was highest in cord blood and progressively declined with age in peripheral blood T CD4+ and CD8+ cells. In contrast, P-glycoprotein function did not vary with age in CD19+ B or CD16+CD56+ natural killer cells. These data suggest that the decline in P-glycoprotein function in T CD4+ and CD8+ lymphocytes as a function of age may contribute to the decrease in T cell cytolytic activity with aging.

  6. Postirradiation Changes of White Blood Cells and Lymphocyte Subpopulations in Cancer Patients

    Energy Technology Data Exchange (ETDEWEB)

    Ahn, Sung Ja; Chung, Woong Ki; Nam, Taek Keun; Nah, Byung Sik; Noh, Young Hee [Chonnam National University College of Medicine, Kwangju (Korea, Republic of)

    1996-03-15

    Purpose : Radiation-induced alteration in the immune function is well known phenomenon in cancer patients. Our purpose is to evaluate the extent of immune suppression immediately after mediastinal or pelvic irradiation, which include significant volume of active bone marrow in adults. Methods and Materials : 48 cancer patients with mediastinal(N=29) and pelvic irradiation(N=19) were the basis of this analysis. Age ranged from 36 to 76 and mean and median value was 57 years, respectively. Sex ratio was 1.3(M:F 27/21). The immunological parameters were the complete blood cell(CBC) with differential cell(D/C) count, T cell subset(CD3, CD4, CD8, CD19), NK cell test(CD16,CD56), and serum immunoglobulin (lgG, lgA, lgM) level. Results : The mean value of white blood cell(WBC) was reduced from 7017 to 4470 after irradiation (p=0.0000). In the differential count, the number of lymphocyte, neutrophil, and basophil was markedly reduced with statistical significance(p<0.01) and the number of monocyte was not changed and, on the contrary, that of eosinophil was increased by irradiation. In the lymphocyte subpopulation analysis, the number of all subpopulations, CD3(T cell), CD4(helper T cell), CD8(suppressor T cell), CD16(NK cell), CD19(B cell) was reduced with statistical significance. The mean ratio of CD4 to CD8 in all patients was 1.09 initially and reduced to 0.99 after radiotherapy(p = 0.34), but the proportional percentage of all subpopulations was not changed except CD19(B cell) after irradiation.In the immunoglobulin study, initial values of lg G, lg A, and lg M were relatively above the normal range and the only lg M was statistically significantly reduced after radiotherapy(p=0.02). Conclusion : Mediastinal and pelvic irradiation resulted in remarkable suppression of lymphocyte count in contrast to the relatively good preservation of other components of white blood cells. But the further study on the functional changes of lymphocyte after radiotherapy may be

  7. Study of p53 protein expression levels from irradiated peripheral blood lymphocytes for biodosimetry

    Energy Technology Data Exchange (ETDEWEB)

    Cavalcanti, M.B.; Fernandes, T.S. [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Dept. de Energia Nuclear; Amaral, A. [Universite Paris XII (UPXII) (France); Melo, J.A. [Centro de Radioterapia de Pernambuco (CERAPE), PE (Brazil); Neves, M.A.B.; Machado, C.G.F, E-mail: maribrayner@yahoo.com.br [Fundacao de Hematologia e Hemoterapia de Pernambuco, PE (Brazil)

    2005-07-01

    Biodosimetry can be defined as the investigation of radioinduced biological effects in order to correlate them with the absorbed dose. Scoring of unstable chromosomal aberrations and micronuclei, from in vitro irradiated peripheral blood lymphocytes, is commonly used for biodosimetry based on cytogenetic analysis. However, this method of analysis is time-consuming, which may represent a pitfall when fast investigation of a possible exposure to ionizing radiation (IR) is needed. The interaction of IR with the living cell can cause injuries in the DNA molecules. However, normal cells possess mechanisms of repair that are capable to correct those damages. During the repair process of the DNA various proteins are expressed. Among these proteins, p53 plays an important role. This protein is a transcription factor that helps in the maintenance of the genomic integrity. p53 protein is found into the cytoplasm in reduced concentrations and has a short average life. However, expression of p53 protein can be induced by DNA harmful radioinduced, which increases the concentration and the average life of this protein, making possible its detection. Thus, the correlation between the increasing of p53 expression and the irradiation may constitute a fast and reliable method of individual monitoring in cases of accidental or suspected exposures to IR. In this context, the objective of this research was to evaluate the p53 protein expression levels from lymphocytes of the human peripheral blood after in vitro irradiation. For this, samples of peripheral blood from healthy individuals were irradiated with known doses. Lymphocytes were separated on ficoll gradient by centrifugation and re-suspended at 1x 10{sub 6}/mL in RPMI medium enriched with fetal calf serum. Hence, lymphocytes were incubated in 5% CO{sub 2} at 37 deg C prior to the methodology of flow cytometry, using intranuclear antigens for the quantification of p53. In this report, the methodology performed and the results

  8. Air pollution exposure during critical time periods in gestation and alterations in cord blood lymphocyte distribution: a cohort of livebirths

    Directory of Open Access Journals (Sweden)

    Herr Caroline EW

    2010-08-01

    Full Text Available Abstract Background Toxic exposures have been shown to influence maturation of the immune system during gestation. This study investigates the association between cord blood lymphocyte proportions and maternal exposure to air pollution during each gestational month. Methods Cord blood was analyzed using a FACSort flow cytometer to determine proportions of T lymphocytes (CD3+ cells and their subsets, CD4+ and CD8+, B lymphocytes (CD19+ and natural killer (NK cells. Ambient air concentrations of 12 polycyclic aromatic hydrocarbons (PAH and particulate matter 2.5 were measured using fixed site monitors. Arithmetic means of these pollutants, calculated for each gestational month, were used as exposure metrics. Data on covariates were obtained from medical records and questionnaires. Multivariable linear regression models were fitted to estimate associations between monthly PAH or PM2.5 and cord blood lymphocytes, adjusting for year of birth and district of residence and, in further models, gestational season and number of prior live births. Results The adjusted models show significant associations between PAHs or PM2.5 during early gestation and increases in CD3+ and CD4+ lymphocytes percentages and decreases in CD19+ and NK cell percentages in cord blood. In contrast, exposures during late gestation were associated with decreases in CD3+ and CD4+ fractions and increases in CD19+ and NK cell fractions. There was no significant association between alterations in lymphocyte distribution and air pollution exposure during the mid gestation. Conclusions PAHs and PM2.5 in ambient air may influence fetal immune development via shifts in cord blood lymphocytes distributions. Associations appear to differ by exposure in early versus late gestation.

  9. Complete blood count and acetylcholinesterase activity of lymphocytes of demyelinated and ovariectomized rats treated with resveratrol.

    Science.gov (United States)

    Martins, Danieli B; Mazzanti, Cinthia M; Costa, Márcio M; França, Raqueli; Pagnoncelli, Marcielen; Maciel, Roberto M; Schmatz, Roberta; Oliveira, Lizielle; Morsch, Vera; Facco, Grasiela; Visentini, Diandra; Mann, Thais; Mazzanti, Alexandre; Lopes, Sonia T A

    2012-12-01

    Resveratrol is a phytoestrogen that has many beneficial actions. This study aimed to evaluate the effect of resveratrol on the complete blood count (CBC) and the acetylcholinesterase (AChE) activity of lymphocytes of ovariectomized rats experimentally demyelinated by ethidium bromide (EB). Forty adult female Wistar rats (60 days, 200-220 g) were divided randomly into five groups (n = 4) to evaluate the demyelination phase and five groups (n = 4) to evaluate the remyelination phase. In each phase, the groups consisted of sham rats-G1; ovariectomized rats, not demyelinated, treated only with vehicle (ethanol 25%)-G2; demyelinated ovariectomized rats treated only with vehicle-G3; ovariectomized rats, not demyelinated, treated with resveratrol-G4; and demyelinated ovariectomized rats treated with resveratrol-G5. Only during the remyelination phase, CBC showed a significant difference (p < 0.05) in the number of monocytes between G2 and G5 groups. In the demyelination phase, there was a significant decrease (p < 0.05) in the AChE activity in the G4 group, while the G5 group was statistically similar to the G1, G2 and G4 groups. In the remyelination phase, there were no significant differences in the AChE activity among the groups. The treatment for 7 days with resveratrol with or without the experimental demyelization with EB appears to influence the AChE activity of lymphocytes, without changing the number of these cells in the circulation. However, in the remyelination phase, there seems to be stabilization in its effect on the lymphocyte AChE activity.

  10. [Double-strand DNA breaks induction and repair in human blood lymphocytes irradiated with adapting dose].

    Science.gov (United States)

    Osipov, A N; Lizunova, E Iu; Vorob'eva, N Iu; Pelevina, I I

    2009-01-01

    Using a DNA-comet assay was shown that irradiation of human blood lymphocytes at G1 cell cycle with a low conditioning dose (5 cGy) induces an adaptive response (AR) manifested in reduction of the double-strand DNA (DSB) amount induced by challenging dose at 10 Gy. 24 h after conditioning irradiation (48 h after PHA addition) in cells irradiated at both conditioning and challenging doses a relative DBS amount was approximately 24% less in comparison to versus a control irradiated at challenging dose only. 48 h after adapting irradiation this index increased to approximately 35%, while 72 h after was decreased to approximately 29%. AR observed by us during 72 h after its induction did not accompanied by statistically significant changes in DBS repair enhancing. It is possible to assume that basic role in AR forming in lymphocytes under experimental conditions used by us playing the processes preventing radiation-induced DBS formation (antioxidant defense system activation, chromatin conformation changes ets).

  11. In-vitro carbofuran induced micronucleus formation in human blood lymphocytes.

    Science.gov (United States)

    Sharma, R K; Rai, D K; Sharma, B

    2012-12-22

    The farmers in general get exposed to different chemicals including pesticides. Many of these compounds are capable of inducing mutations in DNA and lead to several diseases including cancer. Carbofuran is a broad spectrum pesticide and frequently used in agricultural practices in India. In this study we intended to evaluate DNA damage inflicted by pesticide exposure in human blood lymphocytes under in vitro condition. The lymphocytes were exposed to varying concentrations of carbofuran (0—50μM) and analyzed by means of the micronucleus (MN) test. The results obtained showed significant increase in MN frequency after exposure to 5, 10, 25 and 50μM of carbofuran as compared to the control group. The frequencies of MN were observed to be in concentration dependent manner. As we further increase the concentration of carbofuran, we observed significant decrease in the mean percentage of binucleated cells (70—49%) and increase in the number of micronuclei formed per 1000 binucleated cells. Simultaneously, we also observed reduction in Cytokinesis—Block Proliferation index (CBPI) with increase in the carbofuran concentrations. The results indicate that this pesticide may exhibit genotoxic effect at higher concentrations. This study emphasizes the need to reinforce the good practices campaigns in order to enlighten those who work with pesticides and also to make them aware about the importance of using protective measures.

  12. Reference ranges and age-related changes of peripheral blood lymphocyte subsets in Chinese healthy adults

    Institute of Scientific and Technical Information of China (English)

    JIAO Yang; QIU ZhiFeng; XIE Jing; LI DongJing; LI TaiSheng

    2009-01-01

    This study was performed to build region-specific reference ranges of peripheral blood lymphocyte subsets for Chinese healthy adults from the young to the elderly and analyze the trends of changes in lymphocyte subsets for evaluating the impact of age on the values. 151 healthy adults aged 19-86 were recruited based on the SENIEUR protocol. Three sets of reference ranges were finally built applicable for the healthy young (19-44 years), middle-aged (45-64 years) and elder adults (>65). Comparisons in parameters among the three cohorts showed that e statistically significant increase in CD16CD56+ NK cell was observed between the middle-aged and elder cohorts, whereas for the majority of the parameters, a significant decline was observed between the young and the middle-aged cohorts.Further results showed that inverse correlations were observed between the age and CD19+ B, CD3+T,CD3+CD4+1, CD4+CD45RA+CD62L+ naTve T cell and CD4+CD28+/CD4+, while the positive one was identified between the age end the NK cell. These significant changes of the most of immune parameters provided evidence for immunosenescence. Notably, T cell activation markers of CD8+CD38+ and CD8+HLA-DR+ showed reverse trends of association with age, which provides a clue for further researches on the mechanisms underlying the paradoxical clinical presentation of the elder patients.

  13. Genotoxicity evaluation of drinking water sources in human peripheral blood lymphocytes using the comet assay

    Institute of Scientific and Technical Information of China (English)

    WU Yulin; CHEN Haigang; LI Zhaoli; SUN Liwei; QU Mengmeng; LI Mei; KONG Zhiming

    2008-01-01

    The potential harm of organic pollutants in drinking water to human health is widely focused on in the world; more and more pollutants with genotoxic substances are released into the aquatic environment. Water source samples were collected from 7 different localities of Nanjing City. The potential genotoxicity of organic extracts from drinking water sources were investigated by means of the comet assay in human peripheral lymphocytes. The results showed that all the organic extracts from all the water source samples could induce DNA damages of human peripheral blood lymphocytes at different levels. A significant difference (P < 0.01) was observed when compared with the solvent control. The DNA damage increased with the increase of the dosage of the original water source. Significant differences of DNA damage were observed in different drinking water sources, as shown by the multiple comparisons analysis at the dosage of 100×; the degree of DNA damage treated by Hushu waterworks (at town level) was the most serious, the arbitrary units (AU) was 141.62±6.96, however, that of Shangyuanmen waterworks (at city level) was only 109.64±2.97. The analysis also revealed that the genotoxicity of town's water sources was higher than that of the city. The results demonstrated that the comet assay can be successfully applied to the genotoxicity monitoring programs of drinking water sources.

  14. Geno- and cytotoxicity of salinomycin in human nasal mucosa and peripheral blood lymphocytes.

    Science.gov (United States)

    Scherzad, Agmal; Hackenberg, Stephan; Schramm, Carolin; Froelich, Katrin; Ginzkey, Christian; Hagen, Rudolf; Kleinsasser, Norbert

    2015-06-01

    Salinomycin is usually applied in stock breading but has also been described as a promising agent against cancer stem cells (CSC). However, knowledge about the toxicity of this ionophor substance is incomplete. The aim of this study was to investigate cyto- and genotoxic effects of salinomycin in human non-malignant cells. Primary human nasal mucosa cells (monolayer and mini organ cultures) and peripheral blood lymphocytes from 10 individuals were used to study the cytotoxic effects of salinomycin (0.1-175 μM) by annexin-propidiumiodide- and MTT-test. The comet assay was performed to evaluate DNA damage. Additionally, the secretion of interleukin-8 was analyzed by ELISA. Flow cytometry and MTT assay revealed significant cytotoxic effects in nasal mucosa cells and lymphocytes at low salinomycin concentrations of 10-20 μM. No genotoxic effects could be observed. IL-8 secretion was elevated at 5 μM. Salinomycin-induced cytotoxic and pro-inflammatory effects were seen at concentrations relevant for anti-cancer treatment. Concurrent to the evaluation of salinomycin application in experimental oncology, adverse effects in non-malignant cells need to be monitored and reduced as much as possible. Further studies are also warranted to evaluate the toxic effects in a variety of human cell systems, e.g., liver, kidney and muscle cells.

  15. Effect of propolis on mitotic and cellular proliferation indices in human blood lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Montoro, A.; Almonacid, M.; Villaescusa, J. [Valencia Hospital Univ. la Fe, Servicio de Proteccion Radiologica (Spain); Barquinero, J. [Barcelona Univ. Autonom, Servicio de Dosimetria Biologica, Unidad de Antropologia, Dept. de Biologia Animal, Vegetal y Ecologia, barcelona (Spain); Barrios, L. [Barcelona Univ. Autonoma, Dept. de Biologia Celular y Fisiologia. Unidad de Biologia Celular (Spain); Verdu, G. [Valencia Univ. Politecnica, Dept. de Ingenieria Quimica y Nuclear (Spain); Perez, J. [Hospital la Fe, Seccion de Radiofisica, Servicio de Radioterapia, valencia (Spain)

    2006-07-01

    The study of the frequency of chromosomal aberrations per cell is the tool used in Biological dosimetry studies. Using dose-effect calibration curve obtained in our laboratory, we can evaluate the radioprotector effect of the EEP (ethanolic extract of propolis) in cultures in vitro. Propolis is the generic name for resinous substance collected by honeybees. The results showed a reduction in chromosomal aberrations's frequency of up to 50 %. The following study consisted of analyzing human peripheral blood lymphocytes exposed to 2 Gy {gamma} rays, in presence and absence of EEP, the change in the frequency of chromosome aberrations was analysed with biological dosimetry. The protection against the formation of dicentric and ring was dose-dependent, but there seemed to be a maximum protection, i.e. a further increase in the concentration of EEP does not show additional protection. This work studies the effect of the EEP of the cellular cycle using the mitotic and cellular proliferation index, as an alternative for the screening cytostatic activity. The results indicate that the lymphocytes which were cultures in presence of EEP exhibited a significant and dependent-concentration decrease in mitotic index and proliferation kinetics. The possible mechanisms involved in the radioprotective influence of EEP are discussed. (authors)

  16. Setae from larvae of the northern processionary moth (Thaumetopoea pinivora, TP) stimulate proliferation of human blood lymphocytes in vitro.

    Science.gov (United States)

    Holm, Göran; Andersson, Margareta; Ekberg, Monica; Fagrell, Bengt; Sjöberg, Jan; Bottai, Matteo; Björkholm, Magnus

    2014-01-01

    Larvae of the Northern pine processionary moth (Thaumetopoea pinivora, TP) carry microscopic needles (setae), which by penetrating skin and mucous membranes, may cause inflammatory/immune derived symptoms in man. In the present study the stimulatory effects of setae on human blood lymphocytes in vitro was investigated. Blood mononuclear cells were separated from venous blood or buffy coat of ten healthy individuals, six previously exposed to setae and four with no known exposure. Lymphoproliferation was measured as uptake of 3H-thymidine. Setae were prepared from TP larvae. Setae and saline setae extracts stimulated proliferation of T-lymphocytes in the presence of monocytic cells. Stimulation was pronounced in cells from persons who had been exposed to setae, and weak in cells from non-exposed donors. Chitin also induced lymphocyte proliferation in most donors, but to a lesser extent and independently of donor's previous exposure to setae. In conclusion, setae contain molecules that in the presence of monocytes activate human T-lymphocytes to proliferation. The antigenic nature of stimulatory molecules was supported by the significantly stronger lymphocyte response in persons previously exposed to setae than in non-exposed donors. The nature of such molecules remains to be defined.

  17. Setae from larvae of the northern processionary moth (Thaumetopoea pinivora, TP stimulate proliferation of human blood lymphocytes in vitro.

    Directory of Open Access Journals (Sweden)

    Göran Holm

    Full Text Available Larvae of the Northern pine processionary moth (Thaumetopoea pinivora, TP carry microscopic needles (setae, which by penetrating skin and mucous membranes, may cause inflammatory/immune derived symptoms in man. In the present study the stimulatory effects of setae on human blood lymphocytes in vitro was investigated. Blood mononuclear cells were separated from venous blood or buffy coat of ten healthy individuals, six previously exposed to setae and four with no known exposure. Lymphoproliferation was measured as uptake of 3H-thymidine. Setae were prepared from TP larvae. Setae and saline setae extracts stimulated proliferation of T-lymphocytes in the presence of monocytic cells. Stimulation was pronounced in cells from persons who had been exposed to setae, and weak in cells from non-exposed donors. Chitin also induced lymphocyte proliferation in most donors, but to a lesser extent and independently of donor's previous exposure to setae. In conclusion, setae contain molecules that in the presence of monocytes activate human T-lymphocytes to proliferation. The antigenic nature of stimulatory molecules was supported by the significantly stronger lymphocyte response in persons previously exposed to setae than in non-exposed donors. The nature of such molecules remains to be defined.

  18. Dynamics of heat shock protein 70 concentrations in peripheral blood lymphocyte lysates during pregnancy in lactating Holstein-Friesian cows.

    Science.gov (United States)

    Yániz, J L; López-Gatius, F; Almería, S; Carretero, T; García-Ispierto, I; Serrano, B; Smith, R F; Dobson, H; Santolaria, P

    2009-11-01

    The aim of this study was to characterize the dynamics of the concentrations of heat shock protein 70 kDa (HSP70) in peripheral blood lymphocytes of lactating Holstein-Friesian dairy cows (Bos taurus) during pregnancy. The detection of pregnancy was carried out and blood samples collected on Days 40, 90, 120, 150, 180, and 210 of gestation from 46 cows (11 primiparous and 35 pluriparous, 34 seropositive and 12 seronegative to Neospora caninum). Peripheral blood lymphocytes were isolated by density gradient centrifugation. Serologic analysis of Neospora infection and determinations of HSP70 concentrations in lymphocyte lysates were carried out using commercial enzyme-linked immunosorbent assay kits. Climate variables were monitored using on-farm data loggers. Heat shock protein 70 concentrations increased in lymphocytes as gestation progressed, particularly in primiparous cows, with no effect from Neospora infection, climate variables, milk production, semen-providing bull, or outcome of gestation (singletons or twins). Our results show that HSP70 concentrations increased in lymphocytes as gestation progressed and were not affected by stressful factors, such as milk production, heat stress, chronic infection (neosporosis), or twin pregnancies.

  19. LYMPHOCYTE SUBPOPULATIONS OF BLOOD AND PLEURAL EXUDATE IN MBT-NEGATIVE AND MBT-POSITIVE TUBERCULOUS PLEURISY

    Directory of Open Access Journals (Sweden)

    E. A. Jureva

    2010-01-01

    Full Text Available Subpopulation indices of peripheral blood and pleural exudate lymphocytes have been studied in the patients with various clinical variants of a tuberculous pleurisy. It has been shown that, in patients with both MBT-negative and MBT-positive variants of tubercular pleurisy, decreased quantity of CD3- and CD4-positive lymphocytes was observed, whereas an increase in numbers of CD8+ and CD16+ lymphocytes was more pronounced in the persons with MBT-negative pleurisy. In pleural exudates from the patients with tubercular pleurisy, the numbers of CD3+ lymphocytes proved to be significantly higher than their contents in peripheral blood. It may be assumed that lymphocytosis in pleural exudates results from migration of the cells from peripheral blood into pleural cavity. It is revealed that, during exudative pleurisy of tuberculosis origin, T-cells with CD4+ and CD8+ phenotype are more likely subject to migration into pleural cavity, accomplished by additional migration of CD16+ lymphocytes in MBT-positive cases of pleurisy.

  20. Effect of in vitro x-irradiation on human peripheral blood T and B lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Prusek, W. (Szpital Wojewodzki, Wroclaw (Poland)); Astaldi, G. (The Blood Research Foundation Centre, Tortona (Italy))

    1979-01-01

    The effect of in vitro irradiation with increasing in logarythmic progress X-ray doses on lymphocyte viability and on T and B lymphocyte populations was studied in normal adults, patients with myasthenia gravis and in patients undergoing long-term steroid therapy. Decrease in numbers of lymphocytes carrying T or B lymphocyte surface markers was higher than the viable cell loss. The decrease showed no linear correlation with X-ray doses applied, which might reflect the existence of radioresistant T and B lymphocytes. A higher so-called early radiosensitivity of B lymphocytes was demonstrated. In patients with myasthenia gravis early radioresistance of T lymphocytes was detected. In patients undergoing long-term steroid therapy, an increase in numbers of cells lacking markers of any of lymphocyte populations was found in parallel with a decrease in T lymphocyte number which, in these patients, showed a higher radiosensitivity.

  1. Ambiguous nucleus regulates the proliferation and percentage of T lymphocytes in peripheral blood

    Institute of Scientific and Technical Information of China (English)

    Wei Wang; Wei Chen; Yingwu Mei; Bin Guo; Zhanqing Yang; Shoupeng Fu; Zhanpeng Yue; Juxiong Liu

    2011-01-01

    The aim of this study was to examine the immunomodulatory role of the unilateral ambiguous nucleus (Amb). We performed electrical stimulation of the unilateral Amb, electrical stimulation of the left parietal cortex and the lateral hypothalamus following unilateral Amb lesion, as well as microinjection of acetylcholine chloride and hemicholine-3 into the unilateral Amb, and electrical stimulation of the unilateral Amb after injection of atropine, mecamylamine, propranolol, and phentolamine. Results showed that the number and proliferation of peripheral blood T lymphocytes were increased after electrical stimulation of the unilateral Amb. The cholinergic neurons in the Amb released choline substances to alter cellular immunity, thus confirming that the Amb mediates the neuro-immunomodulatory process.

  2. Extended interferon-alpha therapy accelerates telomere length loss in human peripheral blood T lymphocytes.

    Directory of Open Access Journals (Sweden)

    Joel M O'Bryan

    Full Text Available BACKGROUND: Type I interferons have pleiotropic effects on host cells, including inhibiting telomerase in lymphocytes and antiviral activity. We tested the hypothesis that long-term interferon treatment would result in significant reduction in average telomere length in peripheral blood T lymphocytes. METHODS/PRINCIPAL FINDINGS: Using a flow cytometry-based telomere length assay on peripheral blood mononuclear cell samples from the Hepatitis-C Antiviral Long-term Treatment against Cirrhosis (HALT-C study, we measured T cell telomere lengths at screening and at months 21 and 45 in 29 Hepatitis-C virus infected subjects. These subjects had failed to achieve a sustained virologic response following 24 weeks of pegylated-interferon-alpha plus ribavirin treatment and were subsequently randomized to either a no additional therapy group or a maintenance dose pegylated-IFNα group for an additional 3.5 years. Significant telomere loss in naïve T cells occurred in the first 21 months in the interferon-alpha group. Telomere losses were similar in both groups during the final two years. Expansion of CD8(+CD45RA(+CD57(+ memory T cells and an inverse correlation of alanine aminotransferase levels with naïve CD8(+ T cell telomere loss were observed in the control group but not in the interferon-alpha group. Telomere length at screening inversely correlated with Hepatitis-C viral load and body mass index. CONCLUSIONS/SIGNIFICANCE: Sustained interferon-alpha treatment increased telomere loss in naïve T cells, and inhibited the accumulation of T cell memory expansions. The durability of this effect and consequences for immune senescence need to be defined.

  3. CD 13/APN expression in peripheral blood lymphocytes and skin lesions in patients with advanced psoriasis vulgaris

    Institute of Scientific and Technical Information of China (English)

    Liu Taihua; Liu Defang; Chen Yihua; Hu Zonghai; Chen Lu; Luo Chen; Xu Zhejuan

    2009-01-01

    Objective: To observe the expression of CDI3/APN in peripheral blood lymphocytes and skin lesions of patients with advanced psoriasis vulgaris, and discuss its effect on the pathogenesis of psoriasis. Methods: CDI 3 expression in peripheral blood lymphocytes and skin lesions was detected by flow cytometry and immunohistochemical technique, respectively. Results were compared with those of healthy controls. Results: CD13 expression was significantly higher in peripheral blood lymphocytes of patients with advanced psoriasis vulgaris than in that of healthy controls, and in skin lesions than in healthy skin tissues. The expression was mainly in the suprabasal layers of skin lesions, andpositively correlated to PASI (R=0.78029). Conclusion: The significantly higher expression of CDI3 in peripheralblood lymphocytes and skin lesions of the patients with advanced psoriasis vulgaris probably is related to immunological abnormality, blood vessel abnormality and proliferation of keratinocyte in the pathogenic course of psoriasis. It may be a novel and effective way to treat psoriasis with specific CD13 inhibitors.

  4. Achievements and challenges of adoptive T cell therapy with tumor-infiltrating or blood-derived lymphocytes for metastatic melanoma

    DEFF Research Database (Denmark)

    Svane, Inge Marie; Verdegaal, Els M

    2014-01-01

    Adoptive cell therapy (ACT) based on autologous T cell derived either from tumor as tumor-infiltrating lymphocytes (TILs) or from peripheral blood is developing as a key area of future personalized cancer therapy. TIL-based ACT is defined as the infusion of T cells harvested from autologous fresh...... review....

  5. Chromosome aberrations frequency in peripheral blood lymphocytes in young tobacco smoking and non-smoking people

    Directory of Open Access Journals (Sweden)

    Anja Haverić

    2016-10-01

    Full Text Available Introduction: Cigarette smoking is associated with severe health problems, especially cancers. In addition, cigarette smoking causes different genotoxic effects. Chromosome aberrations are one of well-known intermediate end points in carcinogenesis. The aim of this study was to compare frequencies of chromosome aberrations in peripheral blood lymphocytes between young smokers and non-smokes groups.Methods: The study was conducted with 30 smokers (average age 26.93 years and 30 non-smokers (average age 26.96 years, and included the analysis of 100 metaphases per each blood sample. Differences in the arithmetic means of determined frequencies of chromosome aberrations were tested by two-tailed t-test for independent samples with the significance level of p < 0.05.Results: The results showed a significant increase in the frequencies of chromatid-type aberrations and total structural chromosome aberrations in smoker group. Frequencies of numerical aberrations did not differ significantly between two groups.Conclusions: This study confirmed genotoxicity of cigarette smoking and provided new evidence about its clastogenic activity.

  6. Chromosomal Aberrations in Human Peripheral Blood Lymphocytes after Exposure to Ionizing Radiation

    Science.gov (United States)

    Ryu, Tae Ho; Kim, Jin-Hong; Kim, Jin Kyu

    2016-01-01

    Biological dosimetry using chromosome aberration analyses in human peripheral blood lymphocytes is suitable and useful tool for estimating the dose when a nuclear or radiological emergency is investigated. Blood samples from five healthy donors were obtained to establish dose-response calibration curves for chromosomal aberrations after exposure to ionizing radiation. In this work, dicentric assay and CBMN assay were compared considering the sensitivity and accuracy of dose estimation. In a total of 21,688 analyzed metaphase spreads, 10,969 dicentric chromosomes, 563 centric rings and 11,364 acentric chromosomes were found. The number of metaphase cells decreased with increasing radiation dose. The centric rings were not found in the non-irradiated control. There was no relationship between radiation dose and acentric ring induction. The frequency of total MN increased in a dose-dependent manner. In comparison with the control value, MN increased about 9, 32, 75, 87, and 52 fold higher after treatment with 1, 2, 3, 4, and 5 Gy, respectively. The results revealed that the mean frequency of chromosomal aberrations, both in dicentric and in micronuclei analyses increased with increasing radiation dose. PMID:28217281

  7. Reference ranges and age-related changes of peripheral blood lymphocyte subsets in Chinese healthy adults

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    This study was performed to build region-specific reference ranges of peripheral blood lymphocyte subsets for Chinese healthy adults from the young to the elderly and analyze the trends of changes in lymphocyte subsets for evaluating the impact of age on the values.151 healthy adults aged 19-86 were recruited based on the SENIEUR protocol.Three sets of reference ranges were finally built applicable for the healthy young(19-44 years),middle-aged(45-64 years) and elder adults(≥65).Comparisons in parameters among the three cohorts showed that a statistically significant increase in CD16CD56+ NK cell was observed between the middle-aged and elder cohorts,whereas for the majority of the parameters,a significant decline was observed between the young and the middle-aged cohorts.Further results showed that inverse correlations were observed between the age and CD19+ B,CD3+ T,CD3+CD4+ T,CD4+CD45RA+CD62L+ nave T cell and CD4+CD28+/CD4+,while the positive one was identified between the age and the NK cell.These significant changes of the most of immune parameters provided evidence for immunosenescence.Notably,T cell activation markers of CD8+CD38+ and CD8+HLA-DR+ showed reverse trends of association with age,which provides a clue for further researches on the mechanisms underlying the paradoxical clinical presentation of the elder patients.

  8. Effect of copper excess on peripheral blood T-lymphocytes in the chicken

    Institute of Scientific and Technical Information of China (English)

    Cui Hengnmin; Peng Xi; Deng Junliang; Xu Zhiyong; Zhu Kuicheng

    2008-01-01

    Experimental study was conducted to examine the effect of copper excess on the peripheral blood Tlymphocyte by the methods of flow cytometry (FCM) and experimental pathology.420 one-day-old Avian chickens were randomly divided into seven groups, and fed on diets as follows: 1 .controls (Cu 11mg/kg)and 2.copper excess( Cu 100mg/kg, copper excess group Ⅰ; Cu 200mg/kg, copper excess group Ⅱ; Cu 300mg/kg, copper excess group Ⅲ; Cu 400mg/kg, copper excess group Ⅳ; Cu 500mg/kg, copper excess group V;Cu 600mg/kg,copper excess group Ⅵ) for six weeks.The results were as follows: 1) In thymus, lymphocytes in the medulla were decreased in number in copper excess groups Ⅲ, Ⅳ,Ⅴ and Ⅵ,and the increased and enlarged thymic corpuscles and the proliferated reticular cells were also observed in both copper excess group Ⅴ and copper excess group Ⅵ in comparison with those of control group.2) The percentage of CD4 + T cells was markedly decreased from 2 to 6 weeks of age in copper excess groups Ⅳ,Ⅴ and Ⅵ (P<0.05 or P<0.01).3) The percentage of CD8+ T cell was not varied in six copper excess groups during the experiment when compared with that of control group ( P>0.05).4) The CD4+ /CDs + ratio was lower from 2 to 6 weeks of age in copper excess groups Ⅳ, Ⅴ and Ⅵ than in control group (P<0.05 or P<0.01).5) It was concluded that dietary copper in excess of 300rag / kg suppressed the development of T-lymphocytes and reduced the percentage of CD4+ T ceils and the CD4+/CD8+ ratio, and resulted in pathological injury of the thymus.Cellular immune function was finally impaired.

  9. Analysis of the T-Lymphocytes and Their Subpopu-lations of the Peripheral Blood in Patients with Urticaria

    Institute of Scientific and Technical Information of China (English)

    王冬云; 彭振辉; 谭升顺; 楚瑞琦; 刘平

    2003-01-01

    Objective: To study the function of cellular immunity of patients with urticaria.Methods:T-lymphocytes subpopulations of the peripheraal blood in 60 patients with urticaria and 40 henlthy controls were examined by flow cylonuetry. Results: The number of CD3+ and CD4+ cells in the urticaria group were significantly lower than those in the control group ( P<0. 01 ), especially in patients with acute urticaria. Conclusion: There was immunologic dysfunction of T lymphocyte in the patients with urticaria, and not only humoral immunity takes part but also cellular immunity plays a certain role in the pathogenesis of urticaria.

  10. Ex vivo measurement of calpain activation in human peripheral blood lymphocytes by detection of immunoreactive products of calpastatin degradation.

    Directory of Open Access Journals (Sweden)

    Jacek M Witkowski

    2008-01-01

    Full Text Available Limited proteolysis of multiple intracellular proteins by endogenous Ca-dependent cysteine proteases--calpains--is an important regulatory mechanism for cell proliferation, apoptosis etc. Its importance for cellular functions is stressed by existence of endogenous calpain inhibitors--calpastatins. The calpain-calpastatin system within living cells is in a fragile balance, which depends on both partners. The interdependence of calpain--a protease--and calpastatin--an endogenous inhibitor and at the same time a substrate for this enzyme makes any assessment of actual activity of this enzyme in the cells very difficult. In this work we made an attempt to estimate and compare the activity of calpain in human peripheral blood lymphocytes by assessing the levels of limited proteolysis of calpastatin in these cells by western blot, while at the same time the levels of calpain protein inside these cells was measured by flow cytometry. Our results indicate that it is possible to compare (semi-quantitatively the activities of calpain in peripheral blood CD4+ and CD19+ lymphocytes from various donors that way. Preliminary results showed that calpain activity is increased in the CD4+ T cells isolated from peripheral blood of rheumatoid arthritis patients as compared to control lymphocytes. Extremely high intrinsic activity of calpain was detected in chronic lymphocytic leukemia (CD19+ cells. All this confirms the detection of immunoreactive products of calpastatin as a good maker of endogenous calpain activity.

  11. Mitogen-induced lymphocyte proliferation in loggerhead sea turtles: comparison of methods and effects of gender, plasma testosterone concentration, and body condition on immunity.

    Science.gov (United States)

    Keller, Jennifer M; McClellan-Green, Patricia D; Lee, A Michelle; Arendt, Mike D; Maier, Philip P; Segars, Al L; Whitaker, J David; Keil, Deborah E; Peden-Adams, Margie M

    2005-02-10

    A fully functioning immune system is vital to the survival of threatened and endangered sea turtles. Immunological protection against diseases in any organism can be reduced by a number of natural and anthropogenic factors, such as seasonal changes, malnutrition, disease states, and contaminant exposure. These factors are even more critical when they occur in endangered species or populations. To identify alterations in the immunological health of loggerhead sea turtles (Caretta caretta), the mitogen-induced lymphocyte proliferation (LP) assay was developed using peripheral blood leukocytes (PBLs). Collection and culture conditions were optimized for this assay using non-lethal blood samples collected from free-ranging turtles along the southeastern US coast. During the collection, two anticoagulants (sodium heparin and lithium heparin) were compared to determine effects of different ions on assay results. Optimal culture conditions were established for loggerhead PBLs while two different methods of measuring LP were compared: (1) the traditional radioactive (3)H-thymidine assay and (2) a non-radioactive, colorimetric method utilizing 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium (MTT). The results indicate that the (3)H-thymidine and the non-radioactive MTT methods did not correlate with each other and that the use of heparin type did not influence the results of the LP assay. Lastly, using these optimized methods, we investigated the effect of gender, plasma testosterone concentration, and body condition on LP in loggerhead turtles and found that none of the parameters largely influenced LP.

  12. Cancer Risk Estimates from Space Flight Estimated Using Yields of Chromosome Damage in Astronaut's Blood Lymphocytes

    Science.gov (United States)

    George, Kerry A.; Rhone, J.; Chappell, L. J.; Cucinotta, F. A.

    2011-01-01

    To date, cytogenetic damage has been assessed in blood lymphocytes from more than 30 astronauts before and after they participated in long-duration space missions of three months or more on board the International Space Station. Chromosome damage was assessed using fluorescence in situ hybridization whole chromosome analysis techniques. For all individuals, the frequency of chromosome damage measured within a month of return from space was higher than their preflight yield, and biodosimetry estimates were within the range expected from physical dosimetry. Follow up analyses have been performed on most of the astronauts at intervals ranging from around 6 months to many years after flight, and the cytogenetic effects of repeat long-duration missions have so far been assessed in four individuals. Chromosomal aberrations in peripheral blood lymphocytes have been validated as biomarkers of cancer risk and cytogenetic damage can therefore be used to characterize excess health risk incurred by individual crewmembers after their respective missions. Traditional risk assessment models are based on epidemiological data obtained on Earth in cohorts exposed predominantly to acute doses of gamma-rays, and the extrapolation to the space environment is highly problematic, involving very large uncertainties. Cytogenetic damage could play a key role in reducing uncertainty in risk estimation because it is incurred directly in the space environment, using specimens from the astronauts themselves. Relative cancer risks were estimated from the biodosimetry data using the quantitative approach derived from the European Study Group on Cytogenetic Biomarkers and Health database. Astronauts were categorized into low, medium, or high tertiles according to their yield of chromosome damage. Age adjusted tertile rankings were used to estimate cancer risk and results were compared with values obtained using traditional modeling approaches. Individual tertile rankings increased after space

  13. [Peripheral blood T lymphocyte subtypes in multiple sclerosis--dependance of clinical course and duration of the disease].

    Science.gov (United States)

    Vojinović, S; Vojinović, K; Kamenov, B; Vojinović, D; Gocić-Stanković, D

    1994-01-01

    Multiple sclerosis is a disease mediated by immunological mechanisms, with characteristics of an autoimmune prosses. We registered changes in distribution of immunophenotipisation markers CD2, CD3, CD4, CD8, CD56 and DR, by indirect immunoflourescence assay, on immune cells of peripheral blood. We tested 20 patients with clinically definite category of illness, in exacerbation, and 10 healthy individuals. Multiple sclerosis patients had changes in distribution of T cell subtypes in exacerbation, which correlated with clinical course and duration of the disease. Relapsing-remitting course of disease is followed by decrease of activated T lymphocytes and fluctuation of CD4+ T lymphocytes, while there are no changes in studied markers at patients with progressive course. Duration of the disease over 10 years is followed by decreases of CD4+ and CD8+ T lymphocytes, independent of course of the disease.

  14. Metaphase yields from staphylococcal enterotoxin A stimulated peripheral blood lymphocytes of unirradiated and irradiated aged rhesus monkeys

    Science.gov (United States)

    Hill, F. S.; Cox, A. B.; Salmon, Y. L.; Cantu, A. O.; Lucas, J. N.

    1994-01-01

    The mitogen phytohemagglutinin (PHA) works well in both human and cynomolgus monkey (Macaca fascicularis) lymphocyte cultures to stimulate T cell proliferation. T cells from rhesus monkeys (Macaca mulatta) are less responsive than human cells, producing few metaphases when thousands are required, e.g. in biological dosimetry studies. We show that staphylococcal enterotoxin A (SEA), one of the most potent mitogens known, at a concentration of 0.5 microgram/ml stimulated peripheral lymphocytes to grow with a mitotic index (MI) averaging 0.13 metaphases/cell in old, irradiated rhesus macaques. This was significantly greater (p < 0.001) than that produced by PHA (MI < 0.01) in lymphocytes from the same animals. Whole blood was cultured for 96, 120 and 144 h for five irradiated individuals and for two controls. All cells cultured with SEA produced a high MI with a peak response at 120 h whereas the same cultures showed low MI for each PHA stimulated culture.

  15. Relation between clinical mature and immature lymphocyte cells in human peripheral blood and their spatial label free scattering patterns

    Science.gov (United States)

    Zhang, Lu; Zhao, Xin; Zhang, Zhenxi; Zhao, Hong; Chen, Wei; Yuan, Li

    2016-07-01

    A single living cell's light scattering pattern (LSP) in the horizontal plane, which has been denoted as the cell's "2D fingerprint," may provide a powerful label-free detection tool in clinical applications. We have recently studied the LSP in spatial scattering planes, denoted as the cell's "3D fingerprint," for mature and immature lymphocyte cells in human peripheral blood. The effects of membrane size, morphology, and the existence of the nucleus on the spatial LSP are discussed. In order to distinguish clinical label-free mature and immature lymphocytes, the special features of the spatial LSP are studied by statistical method in both the spatial and frequency domains. Spatial LSP provides rich information on the cell's morphology and contents, which can distinguish mature from immature lymphocyte cells and hence ultimately it may be a useful label-free technique for clinical leukemia diagnosis.

  16. [Changes in blood lymphocytes and their subpopulation in patients with myeloblastic leukemia treated with cytostatic agents].

    Science.gov (United States)

    Urasiński, I; Proniewska, M; Schumacher, K

    1979-01-01

    Quantitative determinations of lymphocytas were done in the active period of the disease, immediately after treatment by the COAP schedule and during remission. In 6 patients the determinations were done several times during 20 weeks of maintenance treatment. It was found that independently of the stage of the disease the absolute lymphocyte count and the counts of B and T populations were low, while that of lymphocyte O population was raised. It was observed that the reduced count concerned all 4 subclasses of lymphocytes B, that is those with surface receptors for IgA, IgM, IgG and IgE immunoglobulins. In remission the values of lymphocytes and their T and B subpopulations increased, failing, however, to reach the normal values. This rise was more pronounced in the case of lymphocytes T. Lymphocyte depression in these patients is explained by the authors as due mainly to intensive cytostatic treatment.

  17. Genomic instability and cellular stress in organ biopsies and peripheral blood lymphocytes from patients with colorectal cancer and predisposing pathologies

    Science.gov (United States)

    Lombardi, Sara; Fuoco, Ilenia; di Fluri, Giorgia; Costa, Francesco; Ricchiuti, Angelo; Biondi, Graziano; Nardini, Vincenzo; Scarpato, Roberto

    2015-01-01

    Inflammatory bowel disease (IBD) and polyps, are common colorectal pathologies in western society and are risk factors for development of colorectal cancer (CRC). Genomic instability is a cancer hallmark and is connected to changes in chromosomal structure, often caused by double strand break formation (DSB), and aneuploidy. Cellular stress, may contribute to genomic instability. In colorectal biopsies and peripheral blood lymphocytes of patients with IBD, polyps and CRC, we evaluated 1) genomic instability using the γH2AX assay as marker of DSB and micronuclei in mononuclear lymphocytes kept under cytodieresis inhibition, and 2) cellular stress through expression and cellular localization of glutathione-S-transferase omega 1 (GSTO1). Colon biopsies showed γH2AX increase starting from polyps, while lymphocytes already from IBD. Micronuclei frequency began to rise in lymphocytes of subjects with polyps, suggesting a systemic genomic instability condition. Colorectal tissues lost GSTO1 expression but increased nuclear localization with pathology progression. Lymphocytes did not change GSTO1 expression and localization until CRC formation, where enzyme expression was increased. We propose that the growing genomic instability found in our patients is connected with the alteration of cellular environment. Evaluation of genomic damage and cellular stress in colorectal pathologies may facilitate prevention and management of CRC. PMID:26046795

  18. Ouabain exacerbates activation-induced cell death in human peripheral blood lymphocytes

    Directory of Open Access Journals (Sweden)

    Mabel B. Esteves

    2005-06-01

    Full Text Available Lymphocytes activated by mitogenic lectins display changes in transmembrane potential, an elevation in the cytoplasmic Ca2+ concentrations, proliferation and/or activation induced cell death. Low concentrations of ouabain (an inhibitor of Na+,K+-ATPase suppress mitogen-induced proliferation and increases cell death. To understand the mechanisms involved, a number of parameters were analyzed using fluorescent probes and flow cytometry. The addition of 100nM ouabain to cultures of peripheral blood lymphocytes activated with 5µg/ml phytohemagglutinin (PHA did not modify the increased expression of the Fas receptor or its ligand FasL induced by the mitogen. However, treatment with ouabain potentiated apoptosis induced by an anti-Fas agonist antibody. A synergy between ouabain and PHA was also observed with regard to plasma membrane depolarization. PHA per se did not induce dissipation of mitochondrial membrane potential but when cells were also exposed to ouabain a marked depolarization could be observed, and this was a late event. It is possible that the inhibitory effect of ouabain on activated peripheral blood lymphocytes involves the potentiation of some of the steps of the apoptotic process and reflects an exacerbation of the mechanism of activation-induced cell death.Quando linfócitos são ativados por lectinas mitogênicas apresentam mudanças do potencial de membrana, elevação das concentrações citoplasmáticas de cálcio, proliferação e/ou morte celular induzida por ativação (AICD. Concentrações baixas de ouabaína (um inibidor da Na,K-ATPase suprimem a proliferação induzida por mitógenos e aumentam a morte celular. Para entender os mecanismos envolvidos, uma série de parâmetros foram avaliados usando sondas fluorescentes e citometria de fluxo. A adição de 100nM de ouabaína para culturas de linfócitos de sangue periférico ativadas por fitohemaglutinina (PHA não modificou o aumento de expressão do receptor Fas ou de

  19. Measurement of subgroups of peripheral blood T lymphocytes in patients with severe acute respiratory syndrome and its clinical significance

    Institute of Scientific and Technical Information of China (English)

    唐小平; 尹炽标; 张复春; 付永贵; 陈伟烈; 陈燕清; 王建; 贾卫东; 徐安龙

    2003-01-01

    Objective To investigate the changes of subgroups of peripheral blood T lymphocytes in patients with severe acute respiratory syndrome (SARS) and its clinical significance. Methods Subgroups of blood T lymphocytes in 93 patients with SARS were detected by flow cytometer. The results detected in 64 normal subjects and 50 patients with AIDS served as controls. Results The numbers of CD3+, CD4+, and CD8+ lymphocytes all significantly decreased in acute phase of patients with SARS [(722±533)/μl, (438±353)/μl, (307±217)/μl)] compared with those in normal controls [(1527±470)/μl, (787±257)/μl, (633±280)/μl, all P<0.01)], which was different from what we observed in patients with AIDS who had decreased CD4+[(296±298)/μl] but ncreased CD8+[(818±566)/μl] counts. The counts of CD3+,CD4+, and CD8+ lymphocytes decreased more apparently in patients with severe SARS. All the five patients who died had CD4+ counts less than 200/μl. As the patients' condition improved, CD3+, CD4+, and CD8+ counts gradually returned to normal ranges. Conclusion The damage of cellular immunity is probably an important mechanism of pathogenesis of SARS.

  20. Metallothionein 1 Isoform Gene Expression Induced by Cadmium in Human Peripheral Blood Lymphocytes

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    To study the gene expression of metallothionein 1 (MT-1) isoforms in human peripheral blood lymphocytes (HPBLs). Methods The expression of mRNA representing the seven active MT-1 genes was determined in HPBLs by quantitative RT-PCR before and after exposure to cadmium. Results Basal expressions of MT-1X, and MT-1A in HPBLs were similar to expression of housekeeping gene. In contrast, the basal gene expressions of MT-1H, 1F, 1E, and 1G were a little transcripts in human HPBLs. No signal was detected for MT-1B. There was a sex difference (P<0.05). in basal gene expression of MT-1E. The levels of gene expression of MT-1A, 1E, 1F, 1G, 1H, and 1X increased, but the level of MT-1B did not increase after exposure to cadmium. Conclusions Gene expressions of MT-1G, MT-1H, MT-1F, and MT-1X in HPBLs can be used as a potential biomarker of cadmium exposure.

  1. Derivation of induced pluripotent stem cells from human peripheral blood T lymphocytes.

    Directory of Open Access Journals (Sweden)

    Matthew E Brown

    Full Text Available Induced pluripotent stem cells (iPSCs hold enormous potential for the development of personalized in vitro disease models, genomic health analyses, and autologous cell therapy. Here we describe the generation of T lymphocyte-derived iPSCs from small, clinically advantageous volumes of non-mobilized peripheral blood. These T-cell derived iPSCs ("TiPS" retain a normal karyotype and genetic identity to the donor. They share common characteristics with human embryonic stem cells (hESCs with respect to morphology, pluripotency-associated marker expression and capacity to generate neurons, cardiomyocytes, and hematopoietic progenitor cells. Additionally, they retain their characteristic T-cell receptor (TCR gene rearrangements, a property which could be exploited for iPSC clone tracking and T-cell development studies. Reprogramming T-cells procured in a minimally invasive manner can be used to characterize and expand donor specific iPSCs, and control their differentiation into specific lineages.

  2. Disturbances in lipid second messengers generation by stimulated blood lymphocytes in breast cancer

    Directory of Open Access Journals (Sweden)

    Galstyan H. M.

    2011-10-01

    Full Text Available Aim. The main objective of this study was the comparative investigation of diverse lipid second messenger (LSM generation by human peripheral blood lymphocytes (HPBL at different (5, 10, 30 and 60 s time points of cell co-stimulation by anti-CD3 and anti-CD28 monoclonal antibodies in norm and breast cancer (BC. Methods. Ficoll-Hypaque gradient centrifugation. Results. The data obtained indicate that some mechanisms of LSM generation/utilization in stimulated crude HPBL were significantly altered in BC compared to norm. Particularly, the reliable generation of arachidonyl-1,2-diacylglycerol (1,2-DAG at the initial step (5 s of cell stimulation observed in norm was depressed in BC and reached the value below the basal level in unstimulated cells. It is important that the disturbances in 1,2-DAG formation in HPBL obtained from patients with BC were identical with those observed earlier in other forms of cancer. Conclusions. We conclude that the regularities revealed are common characteristics for all the types of malignancy studied and can be used as additional testing parameters for cancer definition and individual correction of the chemotherapy programs for disease treatment

  3. Mesenchymal stem cells derived from human placenta suppress allogeneic umbilical cord blood lymphocyte proliferation

    Institute of Scientific and Technical Information of China (English)

    Chang Dong LI; Wei Yuan ZHANG; He Lian LI; Xiao Xia JIANG; Yi ZHANG; Pei Hsien TANG; Ning MAO

    2005-01-01

    Human placenta-derived mononuclear cells (MNC) were isolated by a Percoll density gradient and cultured in mesenchymal stem cell (MSC) maintenance medium.The homogenous layer of adherent cells exhibited a typical fibroblastlike morphology,a large expansive potential,and cell cycle characteristics including a subset of quiescent cells.In vitro differentiation assays showed the tripotential differentiation capacity of these cells toward adipogenic,osteogenic and chondrogenic lineages.Flow cytometry analyses and immunocytochemistry stain showed that placental MSC was a homogeneous cell population devoid of hematopoietic cells,which uniformly expressed CD29,CD44,CD73,CD 105,CD166,laminin,fibronectin and vimentin while being negative for expression of CD31,CD34,CD45 and α-smooth muscle actin.Most importantly,immuno-phenotypic analyses demonstrated that these cells expressed class I major histocompatibility complex (MHC-Ⅰ),but they did not express MHC-Ⅱ molecules.Additionally these cells could suppress umbilical cord blood (UCB) lymphocytes proliferation induced by cellular or nonspecific mitogenic stimuli.This strongly implies that they may have potential application in allograft transplantation.Since placenta and UCB are homogeneous,the MSC derived from human placenta can be transplanted combined with hematopoietic stem cells (HSC) from UCB to reduce the potential graft-versus-host disease (GVHD) in recipients.

  4. NAD(P-DEPENDENT DEHYDROGENASE ACTIVITY IN PERIPHERAL BLOOD LYMPHOCYTES OF INFANTS WITH ENLARGEMENT OF PHARYNGEAL TONSILS

    Directory of Open Access Journals (Sweden)

    L. M. Kurtasova

    2014-01-01

    Full Text Available We have observed and examined 57 children 1 to 3 years old diagnosed with enlargement of pharyngeal tonsils. A control group was presented by 35 healthy children. Bioluminescence technique was applied for studying NAD(P-dependent dehydrogenase activity in peripheral blood lymphocytes. Activation of aerobic respiration and increasing activity of pentose phosphate cycle-dependent plastic processes were registered in blood lymphocytes of children with hypertrophic pharyngeal tonsils; along with decreased function of malate-aspartate shunt in energy metabolism of the cells, diminished anaerobic reaction of NADHdependent LDH, lower interaction between Krebs cycle and reactions of amino acid metabolism, and reduced activity of glutathione reductase.

  5. The impact of everolimus versus mycophenolate on blood and lymphocyte cyclosporine exposure in heart-transplant recipients

    DEFF Research Database (Denmark)

    Gustafsson, Finn; Barth, David; Delgado, Diego H

    2009-01-01

    . METHODS: Twelve-hour pharmacokinetic studies of whole-blood and intralymphocytic CsA concentrations were conducted in long-term heart-transplant recipients treated with mycophenolate mofetil (MMF) + CsA (n = 8) and everolimus + CsA (n = 9). RESULTS: There was a highly significant correlation between blood...... CsA C2 levels and blood CsA AUC(0-12) in groups of patients treated with MMF or everolimus (R(2) 0.93 and 0.96, respectively; P MMF (R(2) = 0.98), there was poor correlation...... between whole-blood C2 and lymphocyte AUC(0-12) in patients treated with everolimus (R(2) = 0.24). CONCLUSION: Standard blood CsA levels accurately predict intralymphocytic exposure to CsA in patients concomitantly treated with MMF but not in patients treated with everolimus....

  6. Expressions of programmed death-1 and programmed death ligand on the surface of peripheral blood lymphocytes in patients with tuberculosis

    Institute of Scientific and Technical Information of China (English)

    胥萍

    2014-01-01

    Objective To describe the expressions of programmed death-1(PD-1)and its ligand PD-L1 on the surface of peripheral blood lymphocytes in patients with tuberculosis.Methods A total of 77 cases of pulmonary tuberculosis were recruited,of which 27 were single infection,41 were coincident with bacterial or fungal infection and 9 patients with diabetes millitus.Twenty-nine

  7. The Dopaminergic System in Peripheral Blood Lymphocytes: From Physiology to Pharmacology and Potential Applications to Neuropsychiatric Disorders

    OpenAIRE

    Buttarelli, Francesca R.; Fanciulli, Alessandra; Pellicano, Clelia; Pontieri, Francesco E.

    2011-01-01

    Besides its action on the nervous system, dopamine (DA) plays a role on neural-immune interactions. Here we review the current evidence on the dopaminergic system in human peripheral blood lymphocytes (PBL). PBL synthesize DA through the tyrosine-hydroxylase/DOPA-decarboxylase pathway, and express DA receptors and DA transporter (DAT) on their plasma membrane. Stimulation of DA receptors on PBL membrane contributes to modulate the development and initiation of immune responses under physiolog...

  8. Correlation between T lymphocyte subsets in peripheral blood lymphocytes and 2-year all-cause mortality in an apparently healthy elderly Chinese cohort

    Institute of Scientific and Technical Information of China (English)

    WANG Yu-hong; ZHANG Jing-yu; QIAO Fang-fang; ZHU Jing; YIN Feng; HAN Hui

    2012-01-01

    Background Few data have been acquired on the predictive value of age-related T-lymphocyte subsets among older individuals.The present study has determined the distribution of T-cell phenotypes and their correlation to 2-year mortality in a cohort of Chinese male seniors.Methods A total of 101 asymptomatic elderly individuals with laboratory homeostasis were enrolled at baseline.Three age subgroups were categorized as young (65-74 years old),middle (75-84 years old ),and old (≥85 years) for age-related comparison.T-cell subsets in peripheral blood were measured by multi-colored flow cytometry.Results At baseline,there was a mild negative correlation by age for total lymphocytes and CD3+ T-cells.The frequency of CD28 and CD95 demonstrated a "curved" rather than linear tendency by age.At 2-year follow-up,little change of T-cell distribution was found among those who remained alive (as survivors) comparing the data at baseline to the 2-year time point.Immune risk phenotypes were distinctly demonstrated between survivors and non-survivors.Conclusions Since few studies have studied on the distribution of T-lymphocyte subsets in an elderly Chinese population,our results have not only provided reference values of T-subsets for aged Chinese men,but confirmed the immune risk phenotypes among elderly Chinese.The inappropriate age-dependent trajectory of CD28-/CD8+ and CD95-/CD8+ by age,which suggested 85 might be an inflexion point of age during T-cell ageing,warrants further exploration of the underlying mechanisms of T-cell ageing.

  9. Exposure to Brominated Trihalomethanes in Water During Pregnancy and Micronuclei Frequency in Maternal and Cord Blood Lymphocytes

    Science.gov (United States)

    Pedersen, Marie; Patelarou, Evridiki; Decordier, Ilse; Vande Loock, Kim; Chatzi, Leda; Espinosa, Ana; Fthenou, Eleni; Nieuwenhuijsen, Mark J.; Gracia-Lavedan, Esther; Stephanou, Euripides G.; Kirsch-Volders, Micheline; Kogevinas, Manolis

    2013-01-01

    Background: Water disinfection by-products have been associated with an increased cancer risk. Micronuclei (MN) frequency in lymphocytes is a marker of genomic damage and can predict adult cancer risk. Objective: We evaluated maternal exposure to drinking water brominated trihalomethanes (BTHM) in relation to MN frequency in maternal and cord blood lymphocytes. Methods: MN frequency was examined in 214 mothers and 223 newborns from the Rhea mother–child cohort in Crete, Greece, in 2007–2008. Residential BTHM water concentrations were estimated during pregnancy using tap water analyses and modeling. Questionnaires on water related habits were used to estimate BTHM exposure from all routes. Associations between BTHM and MN frequency were estimated using negative binomial regression. Results: BTHM concentrations in residential tap water during pregnancy ranged from 0.06 to 7.1 μg/L. MN frequency in maternal binucleated lymphocytes was found to increase with BTHM concentrations in residential water for exposure during the first [rate ratio (RR) for 1 μg/L = 1.05; 95% CI: 1.00, 1.11] and second trimesters (RR for 1 μg/L = 1.03; 95% CI: 1.00, 1.06), and through all routes of BTHM exposure during the first trimester (RR for 1 μg/week = 3.14; 95% CI: 1.16, 8.50). Conclusions: These findings suggest that exposure to BTHM may increase the frequency of MN in maternal binucleated lymphocytes. Citation: Stayner LT, Pedersen M, Patelarou E, Decordier I, Vande Loock K, Chatzi L, Espinosa A, Fthenou E, Nieuwenhuijsen MJ, Gracia-Lavedan E, Stephanou EG, Kirsch-Volders M, Kogevinas M. 2014. Exposure to brominated trihalomethanes in water during pregnancy and micronuclei frequency in maternal and cord blood lymphocytes. Environ Health Perspect 122:100–106; http://dx.doi.org/10.1289/ehp.1206434 PMID:24184846

  10. T Lymphocytes and Inflammatory Mediators in the Interplay between Brain and Blood in Alzheimer's Disease: Potential Pools of New Biomarkers

    Science.gov (United States)

    Mietelska-Porowska, Anna

    2017-01-01

    Alzheimer's disease (AD) is a chronic neurodegenerative disorder and the main cause of dementia. The disease is among the leading medical concerns of the modern world, because only symptomatic therapies are available, and no reliable, easily accessible biomarkers exist for AD detection and monitoring. Therefore extensive research is conducted to elucidate the mechanisms of AD pathogenesis, which seems to be heterogeneous and multifactorial. Recently much attention has been given to the neuroinflammation and activation of glial cells in the AD brain. Reports also highlighted the proinflammatory role of T lymphocytes infiltrating the AD brain. However, in AD molecular and cellular alterations involving T cells and immune mediators occur not only in the brain, but also in the blood and the cerebrospinal fluid (CSF). Here we review alterations concerning T lymphocytes and related immune mediators in the AD brain, CSF, and blood and the mechanisms by which peripheral T cells cross the blood brain barrier and the blood-CSF barrier. This knowledge is relevant for better AD therapies and for identification of novel biomarkers for improved AD diagnostics in the blood and the CSF. The data will be reviewed with the special emphasis on possibilities for development of AD biomarkers.

  11. Estrogen protects the blood-brain barrier from inflammation-induced disruption and increased lymphocyte trafficking.

    Science.gov (United States)

    Maggioli, E; McArthur, S; Mauro, C; Kieswich, J; Kusters, D H M; Reutelingsperger, C P M; Yaqoob, M; Solito, E

    2016-01-01

    Sex differences have been widely reported in neuroinflammatory disorders, focusing on the contributory role of estrogen. The microvascular endothelium of the brain is a critical component of the blood-brain barrier (BBB) and it is recognized as a major interface for communication between the periphery and the brain. As such, the cerebral capillary endothelium represents an important target for the peripheral estrogen neuroprotective functions, leading us to hypothesize that estrogen can limit BBB breakdown following the onset of peripheral inflammation. Comparison of male and female murine responses to peripheral LPS challenge revealed a short-term inflammation-induced deficit in BBB integrity in males that was not apparent in young females, but was notable in older, reproductively senescent females. Importantly, ovariectomy and hence estrogen loss recapitulated an aged phenotype in young females, which was reversible upon estradiol replacement. Using a well-established model of human cerebrovascular endothelial cells we investigated the effects of estradiol upon key barrier features, namely paracellular permeability, transendothelial electrical resistance, tight junction integrity and lymphocyte transmigration under basal and inflammatory conditions, modeled by treatment with TNFα and IFNγ. In all cases estradiol prevented inflammation-induced defects in barrier function, action mediated in large part through up-regulation of the central coordinator of tight junction integrity, annexin A1. The key role of this protein was then further confirmed in studies of human or murine annexin A1 genetic ablation models. Together, our data provide novel mechanisms for the protective effects of estrogen, and enhance our understanding of the beneficial role it plays in neurovascular/neuroimmune disease.

  12. Abnormal humoral immune responses in peripheral blood lymphocyte cultures of bone marrow transplant recipients.

    Science.gov (United States)

    Pahwa, S G; Pahwa, R N; Friedrich, W; O'Reilly, R J; Good, R A

    1982-01-01

    The present study was aimed at investigating recovery of humoral immunity in vitro after bone marrow transplantation in patients with acute leukemia and severe aplastic anemia. Hemolytic plaque assays were utilized to quantitate pokeweed mitogen-stimulated polyclonal immunoglobulin production and sheep erythrocyte antigen-specific antibody responses in cultures of peripheral blood mononuclear cells of 39 patients beginning at 1 month, for variable periods up to a maximum of 4 years after marrow transplantation. Three phases were identified: an early period of primary B cell dysfunction with concomitant immunoregulatory T cell abnormalities--i.e., decreased helper and increased suppressor activities; an intermediate phase in which B cell dysfunction could be attributed in large measure to immunoregulatory T cell abnormalities; and a late phase of normal B and T lymphocyte functions. Patients with graft-versus-host disease differed from those without it in that they often did not manifest increased T cell suppressor activity in the early period, and they were noted to have prolonged and profound B and T cell abnormalities in the chronic phase of their disease. In selected patients, simultaneous assessment of ratios of Leu-2 to Leu-3 antigens on T cells by monoclonal antibodies and of immunoregulatory T cell functions revealed a correlation between the two only late in the post-transplant period. These studies provide an insight into the ontogeny of B cell function in the post-transplant period and indicate that in certain situations phenotypic alterations in T cell subsets cannot reliably be used to predict abnormalities in their function in recipients of marrow transplantation. Images PMID:6211673

  13. [Regularities of endogenous lipid metabolites formation in phorbol 12-miristate 13-acetate-stimulated peripheral blood lymphocytes at leukemia].

    Science.gov (United States)

    Batikian, T B; Akopian, G V; Lazian, M P; Torgomian, T R; Kazarian, R A; Amirkhanian, E S; Tadevosian, Iu V

    2011-01-01

    Regularities of biologically active lipid metabolites formation in dynamics (5, 10, 30, 60 s) by phorbol 12-miristate 13-acetate stimulation in [14C]palmitic acid have been investigated in normal and leukemia peripheral blood lymphocytes prelabeled with [14C]palmitate. In normal cells there was two-phase formation of 1,2-diacylglycerol (5, 30 s), lysophosphatidylcholine (10, 60 s), as well as free palmitic acid at 10 s of stimulation. Under the identical experimental conditions there was inhibition of investigated lipid release processes at early (5 and 10 s) stages of stimulation of leukemic lymphocytes. At later (30, 60 s) terms of these lymphocytes the activation, basically, similar to norm changes in the formation of palmitic acid-containing metabolites except free palmitic acid (the level of which raised only at 60 second of the post-stimulation) was found. Various protein kinases C are involved in the regulation of investigated lipid levels at certain stages of signal transduction both in norm, and in blast cells. Short-term (5, 10 s) activations of healthy donors lymphocytes are coupled to functioning of Ca2+-independent isoforms of protein kinase C. The inhibition of this protein kinase C in leukemic cells leads to normalization of the investigated lipid release. The data obtained suggests disorders of early membrane-bound reactions in agonist - and a protein kinase C-mediated processes of formation palmitic acid-containing lipid metabolites in the leukemic cells in comparison with the norm.

  14. Estimating the number of hematopoietic or lymphoid stem cells giving rise to clonal chromosome aberrations in blood T lymphocytes.

    Science.gov (United States)

    Nakano, M; Kodama, Y; Ohtaki, K; Itoh, M; Awa, A A; Cologne, J; Kusunoki, Y; Nakamura, N

    2004-03-01

    Quantifying the proliferative capacity of long-term hematopoietic stem cells in humans is important for bone marrow transplantation and gene therapy. Obtaining appropriate data is difficult, however, because the experimental tools are limited. We hypothesized that tracking clonal descendants originating from hematopoietic stem cells would be possible if we used clonal chromosome aberrations as unique tags of individual hematopoietic stem cells in vivo. Using FISH, we screened 500 blood T lymphocytes from each of 513 atomic bomb survivors and detected 96 clones composed of at least three cells with identical aberrations. The number of clones was inversely related to their population size, which we interpreted to mean that the progenitor cells were heterogeneous in the number of progeny that they could produce. The absolute number of progenitor cells contributing to the formation of the observed clones was estimated as about two in an unexposed individual. Further, scrutiny of ten clones revealed that lymphocyte clones could originate roughly equally from hematopoietic stem cells or from mature T lymphocytes, thereby suggesting that the estimated two progenitor cells are shared as one hematopoietic stem cell and one mature T cell. Our model predicts that one out of ten people bears a non- aberrant clone comprising >10% of the total lymphocytes, which indicates that clonal expansions are common and probably are not health-threatening.

  15. The effects of biological and life-style factors on baseline frequencies of chromosome aberrations in human lymphocytes

    Directory of Open Access Journals (Sweden)

    Hilada Nefic

    2014-01-01

    Full Text Available Objective: This study investigated the influence of sex and ageing on chromosomal damage and the role of life-style habits on the frequency of chromosomal aberrations (CAs in peripheral blood lymphocytes (PBLs of healthy Bosnian subjects. Materials and Methods: Peripheral blood samples were obtained from 100 healthy, unrelated individuals in Bosnia and Herzegovina during 2010 and 2011. Chromosome preparations were made by dropping and air drying and slides were stained with 10% Giemsa solution (pH 6.8. The cytogenetic analysis was carried out in a cytogenetic laboratory in the Department of Biology of the Faculty of Science in Sarajevo. The category of total structural CAs was sub classified as chromosome-type aberrations (CSAs and chromatid-type aberrations (CTAs while the category of total numerical CAs was sub classified as aneuploid and polyploid mitoses. All statistical analyses were carried out using Microsoft Excel 2010 (Microsoft Corporation and the Windows Kwikstat Winks SDA 7.0.2 statistical software package (Texa Soft Cedar Hill, Texas. Results: Cytogenetic analysis revealed the average number of structural CAs was 2.84 and of numerical CAs was 9.56. There was a significant increase in the frequency of chromosome-type aberrations (1.92 compared with chromatid-type aberrations (CTAs (0.92 and a significant increase in the frequency of aneuploid (8.83 compared with polyploid (0.73 mitoses. Significant positive correlations between age and CTAs in human PBLs were also demonstrated. Additional statistical analysis showed that ageing increase number of numerical CAs in lymphocytes of drinkers. The frequency of structural CAs of females exposed to radiation was significantly greater than in males. Analysis indicates the presence of a positive association between CAs and smoking in younger subjects but a negative correlation between aberrant cells frequencies and alcohol in older drinkers. Conclusion: The results of the study support the

  16. The function of T-lymphocyte subtypes of blood in patients with hyper-IgE syndrome

    Institute of Scientific and Technical Information of China (English)

    LEI Xiao-bing; TAN Sheng-shun; ZENG Wei-hui; WANG Jun-min; ZHANG Pan-jian; YUAN Yuan

    2005-01-01

    Objective: To study the function in cellular immunity of patients with hyper-IgE syndrome (HIE). Methods: T-lymphocyte subtypes of the peripheral blood and cutaneous delayed-type hypersensitivity (DTH) to two recall antigens, tetanus toxoid (TT) and purified protein derivative(PPD), were measured in 5 patients with HIE and 15 healthy controls, respectively. Results: The CD4+ cell counts in HIE group were significantly lower than those in the control group (P<0.01). In contrast, CD8+ cells were significantly higher in HIE group than those in the controls. The induration sizes of DTH to two recall antigens were smaller in HIE group than those in controls (P<0.01). Conclusion: There is an immunologic dysfunction of T lymphocytes in the patients with HIE and T cells play an important role in the pathogenesis.

  17. Peculiarities of induction and persistence of hidden chromosome instability in peripheral blood lymphocytes of persons occupationally exposed to ionizing radiation.

    Science.gov (United States)

    Pilinska, M A; Dybsky, S S; Dybska, O B; Shvayko, L I; Sushko, V O

    2014-09-01

    Objective - to investigate the induction of hidden chromosome instability in persons occupationally exposed to ionizing radiation and its persistence in vitro in successive mitoses. Materials and methods. Using two tests ("G2-bleomycin sensitivity assay" and two-term cultivation of human peripheral blood lymphocytes) voluntary cytogenetic examination of 15 individuals participated in the conversion of the "Shelter" ("Chornobyl NPP") into ecologically safe system had been carried out. Total 24 034 metaphase had been analyzed, of which 12 243 - without additional mutagenic exposure, 11 791 - exposed to bleomycin in vitro at concentration of 0.05 μg/ml. Results. The magnitude and dynamics of background as well as bleomycin-induced cytogenetic effects in both terms of lymphocytes' cultivation in occupational group differed significantly from the group of comparison towards increasing of chromosome instability indices with significant interindividual fluctuations. Conclusion. Interindividual differences in persistence of radiation-induced hidden chromosome instability in successive generations of human somatic cells had been found.

  18. Antiviral activity of derivatized dextrans on HIV-1 infection of primary macrophages and blood lymphocytes.

    Science.gov (United States)

    Seddiki, N; Mbemba, E; Letourneur, D; Ylisastigui, L; Benjouad, A; Saffar, L; Gluckman, J C; Jozefonvicz, J; Gattegno, L

    1997-11-28

    The present study demonstrates at the molecular level that dextran derivatives carboxymethyl dextran benzylamine (CMDB) and carboxymethyl dextran benzylamine sulfonate (CMDBS), characterized by a statistical distribution of anionic carboxylic groups, hydrophobic benzylamide units, and/or sulfonate moieties, interact with HIV-1 LAI gp120 and V3 consensus clades B domain. Only limited interaction was observed with carboxy-methyl dextran (CMD) or dextran (D) under the same conditions. CMDBS and CMDB (1 microM) strongly inhibited HIV-1 infection of primary macrophages and primary CD4+ lymphocytes by macrophage-tropic and T lymphocyte-tropic strains, respectively, while D or CMD had more limited effects on M-tropic infection of primary macrophages and exert no inhibitory effect on M- or T-tropic infection of primary lymphocytes. CMDBS and CMDB (1 microM) had limited but significant effect on oligomerized soluble recombinant gp120 binding to primary macrophages while they clearly inhibit (> 50%) such binding to primary lymphocytes. In conclusion, the inhibitory effect of CMDB and the CMDBS, is observed for HIV M- and T-tropic strain infections of primary lymphocytes and macrophages which indicates that these compounds interfere with steps of HIV replicative cycle which neither depend on the virus nor on the cell.

  19. Monoclonal B lymphocytes with the characteristics of "indolent" chronic lymphocytic leukemia are present in 3.5% of adults with normal blood counts.

    Science.gov (United States)

    Rawstron, Andy C; Green, Michael J; Kuzmicki, Anita; Kennedy, Ben; Fenton, James A L; Evans, Paul A S; O'Connor, Sheila J M; Richards, Stephen J; Morgan, Gareth J; Jack, Andrew S; Hillmen, Peter

    2002-07-15

    Molecular and cellular markers associated with malignant disease are frequently identified in healthy individuals. The relationship between these markers and clinical disease is not clear, except where a neoplastic cell population can be identified as in myeloma/monoclonal gammopathies of undetermined significance (MGUS). We have used the distinctive phenotype of chronic lymphocytic leukemia (CLL) cells to determine whether low levels of these cells can be identified in individuals with normal complete blood counts. CLL cells were identified by 4-color flow cytometric analysis of CD19/CD5/CD79b/CD20 expression in 910 outpatients over 40 years old. These outpatients were age- and sex-matched to the general population with normal hematologic parameters and no evident history of malignant disease. CLL phenotype cells were detectable in 3.5% of individuals at low level (median, 0.013; range, 0.002- 1.458 x 10(9) cells/L), and represented a minority of B lymphocytes (median, 11%; range, 3%-95%). Monoclonality was demonstrated by immunoglobulin light-chain restriction in all cases with CLL phenotype cells present and confirmed in a subset of cases by consensus-primer IgH-polymerase chain reaction. As in clinical disease, CLL phenotype cells were detected with a higher frequency in men (male-to-female ratio, 1.9:1) and elderly individuals (2.1% of 40- to 59-year-olds versus 5.0% of 60- to 89-year-olds, P =.01). The neoplastic cells were identical to good-prognosis CLL, being CD5+23+20(wk)79b(wk)11a(-)22(wk)sIg(wk)CD38-, and where assessed had a high degree (4.8%-6.6%) of IgH somatic hypermutation. The monoclonal CLL phenotype cells present in otherwise healthy individuals may represent a very early stage of indolent CLL and should be useful in elucidating the mechanisms of leukemogenesis.

  20. Dose assessment by quantification of chromosome aberrations and micronuclei in peripheral blood lymphocytes from patients exposed to gamma radiation

    Energy Technology Data Exchange (ETDEWEB)

    Silva-Barbosa, Isvania; Pereira-MagnataI, Simey; Amaral, Ademir [Pernambuco Univ., Recife, PE (Brazil). Dept. de Energia Nuclear. Grupo de Estudos em Radioprotecao e Radioecologia - GERAR; Sotero, Graca [Fundacao de Hematologia e Hemoterapia, Recife, PE (Brazil); Melo, Homero Cavalcanti [Hospital do Cancer, Recife, PE (Brazil). Centro de Radioterapia de Pernambuco]. E-mail: isvania@uol.com.br

    2005-07-15

    Scoring of unstable chromosome aberrations (dicentrics, rings and fragments) and micronuclei in circulating lymphocytes are the most extensively studied biological means for estimating individual exposure to ionizing radiation (IR), which can be used as complementary methods to physical dosimetry or when the latter cannot be performed. In this work, the quantification of the frequencies of chromosome aberrations and micronuclei were carried out based on cytogenetic analyses of peripheral blood samples from 5 patients with cervical uterine cancer following radiotherapy in order to evaluate the absorbed dose as a result of partial-body exposure to 60Co source. Blood samples were collected from each patient in three phases of the treatment: before irradiation, 24 h after receiving 0.08 Gy and 1.8 Gy, respectively. The results presented in this report emphasize biological dosimetry, employing the quantification of chromosome aberrations and micronuclei in lymphocytes from peripheral blood, as an important methodology of dose assessment for either whole or partial-body exposure to IR.

  1. Distribution of natural killer cells and T-lymphocyte subsets in peripheral blood, gallbladder cancer and surrounding tissue

    Institute of Scientific and Technical Information of China (English)

    Gang Liu; Hong Ren; Xue-Jun Sun; Jing-Sen Shi

    2007-01-01

    BACKGROUND:The patient with malignant tumor always show immunologic function drawback and ingravescent with tumor development, especially in the aspect of cell-mediated immunity. This study was undertaken to deifne the relationship between the immune function of local cells and cancer development by investigating the distribution of natural killer (NK) cells and T-lymphocyte subsets in peripheral blood, the cancer tissue and the tissue surrounding gallbladder carcinoma. METHODS:The numbers of CD4+and CD8+T-lymphocytes and NK cells were measured by lfow cytometry in samples taken from gallbladder cancer tissue, the surrounding tissues and peripheral blood of 38 patients, and compared with the numbers in the peripheral blood and gallbladder tissue of 30 patients with cholecystitis as controls. RESULTS:The numbers of CD4+and CD8+T-cells and NK cells in gallbladder cancer tissues were signiifcantly higher than those in the surrounding tissue and gallbladder with gallstone. However, the ratio of CD4+/CD8+was lower in the cancer tissue than that in the surrounding tissue and tissue from gallbladders with gallstones. The distribution of CD4+ and CD8+ T-cells and NK cells in mucous membrane of cholecystitis gallbladder and that in the tissue surrounding gallbladder cancer were signiifcantly different. CONCLUSIONS:Disproportionate and imbalanced distri-bution of NK cells and subsets of T-lymphocytes occurs in the mucous membrane proper of gallbladder cancer and surrounding tissue. Although gallbladder cancer tissue has higher expressions of CD4+, CD8+and NK cells, the immune function is low or in an inhibited state. In gallbladder cancer immunization therapy, local cellular immunological function should be enhanced and the protective barrier improved.

  2. The prognostic role of blood lymphocyte subset distribution in patients with resected high-risk primary or regionally metastatic melanoma

    DEFF Research Database (Denmark)

    Hernberg, Micaela; Mattila, Petri S; Rissanen, Marjo;

    2007-01-01

    Cooperative Group adjuvant interferon study. The frequencies of peripheral blood lymphocyte subsets were monitored by flow cytometry using CD3, CD4, CD8, CD56, and CD69 monoclonal antibodies. Patients with low proportions of CD3+CD4+CD69+ cells and of CD3+CD56+ cells before treatment had an improved disease...... independent prognostic factors for overall survival. Our data show that both the proportions of CD3+CD4+CD69+ cells and of CD3+CD56+ cells seem to have a prognostic potential in the natural course of melanoma. These results need to be confirmed in larger studies. Udgivelsesdato: 2007-Oct...

  3. Development of a preliminary diagnostic measure for bovine leukosis in dairy cows using peripheral white blood cell and lymphocyte counts

    Science.gov (United States)

    NISHIIKE, Masao; HAOKA, Michiyo; DOI, Takashi; KOHDA, Tomoko; MUKAMOTO, Masafumi

    2016-01-01

    Analysis of the association between antibodies against bovine leukemia virus (BLV), BLV proviral load, and white blood cell (WBC) and lymphocyte counts was performed with 774 dairy cows. The average age, WBC counts and lymphoid cell counts tended to be higher in BLV antibody-positive cows than in antibody-negative cows. There was a similar trend in levels of proviral DNA. We analyzed age, WBC counts and lymphocyte counts by principal component analyses to create a distribution chart of the principle component scores. Using the chart, we categorized cows into four quadrants based on additional information, such as the presence of antibody and the levels of proviral DNA. Antibody-positive cows and cows with high BLV proviral load were found mostly in one quadrant of the chart, indicating that it is possible to predict the risk of infection without any knowledge on antibody status by using information, such as WBC counts as a biomarker. When only antibody-positive cows were included in the analysis, a characteristic distribution of different levels of proviral DNA was seen in the quadrants, suggesting that it is possible to estimate the extent of bovine leukosis infection by using this analysis. For this analysis and categorization of the cows into quadrants, we computed a mathematical formulation using discriminant analysis based on age and WBC and lymphocyte counts. This mathematical formulation for the hematological preliminary diagnosis of the disease is recommended as a screening tool to monitor bovine leukosis. PMID:27064146

  4. Metaphase yields from staphylococcal enterotoxin A stimulated peripheral blood lymphocytes of unirradiated and irradiated aged rhesus monkeys

    Energy Technology Data Exchange (ETDEWEB)

    Hill, F.S.; Cantu, A.O.; Lucas, J.N. (Lawrence Livermore National Lab., CA (United States)); Cox, A.B.; Salmon, Y.L. (Air Force Armstrong Lab., Brookes AFB, TX (United States))

    1994-10-01

    The mitogen phytohemagglutinin (PHA) works well in both human and cynomolgus monkey (Macaca fascicularis) lymphocyte cultures to stimulate T cell proliferation. T cells from rhesus monkeys (Macaca mulatta) are less responsive than human cells, producing few metaphases when thousands are required, e.g. in biological dosimetry studies. We show that staphylococcal enterotoxin A (SEA), one of the most potent mitogens known, at a concentration of 0.5 [mu]g/ml stimulated peripheral lymphocytes to grow with a mitotic index (MI) averaging 0.13 metaphases/cell in old, irradiated rhesus macaques. This was significantly greater (p < 0.001) that that produced by PHA (M1<0.01) in lymphocytes from the same animals. Whole blood was cultured for 96, 120 and 144 h for five irradiated individuals and for two controls. All cells cultured with SEA produced a high MI with a peak response at 120 h whereas the same cultures showed low MI for each PHA stimulated culture. (author).

  5. T lymphocytes in the lesional skin and the levels of peripheral blood cytokines in patients with psoriasis

    Directory of Open Access Journals (Sweden)

    İbrahim Kökçam

    2011-03-01

    Full Text Available In the current study, it was aimed to investigate the roles of tissue cellular immunity and serum levels of cytokines in the patients with plaque psoriasis treated with calcipotriol-betamethasone dipropionate.Materials and methods: The study included 20 patients with psoriasis. Peripheral blood and biopsy samples were collected from lesional and normal skins before and after treatment. The results were compared with each other.Results: Immunohistochemical examination revealed significant elevations of CD4+, CD8+ and CD25+ T lymphocytes in the lesional tissues when compared to that in the healthy tissues and post treatment tissue (p0.05. The levels of IL–4, IL–10, TNF-α, IFN-γ and TGF-β1 in serum were not significantly different between before and after treatment periods (p>0.05.Conclusion: Our study demonstrated that there were infiltration of CD4+ and CD8+ cell in the lesional skin and CD8+ T-lymphocytes were the dominant cell types. The improvement of the lesions and significant decreases in CD4+ and CD8+ T-cells in accordance with the treatment strongly support the hypothesis that Th lymphocytes may have prominent roles in the immunopathogenesis of the disease. However, our findings showed that sufficient T-cells still remains in the tissue, which is consistent with the chronic characteristic of the disease, and the topical treatment could not be able to prevent the activation of the disease.

  6. Expression of HLA class Ⅰ and Ⅱ on peripheral blood lymphocytes in HBV infection

    Institute of Scientific and Technical Information of China (English)

    WANG Chuan-xin; WANG Jin-feng; LIU Min; ZOU Xiong; YU Xiu-ping; YANG Xiao-jing; ZHENG Gui-xi

    2006-01-01

    @@ Persistent hepatitis B virus (HBV) infection is the most important reason for chronic hepatitis B,hepatic cirrhosis, and hepatocellular carcinoma.1 T lymphocytes, including CD4+ and CD8+ T cells, are major composition of host cellular immunity.Furthermore, CD8+ cells play a primary role in host immune reaction of anti-tumor and anti-infection.

  7. Cytotoxic T lymphocyte antigen-4 (CTLA-4 A49G polymorphism and autoimmune blood diseases

    Directory of Open Access Journals (Sweden)

    Faruk Aktürk

    2010-06-01

    Full Text Available Objective: The cytotoxic T lymphocyte associated antigen-4 (CTLA-4 is expressed on T lymphocytes, and inhibits the T-cell responses. In animal models, it has been shown that complete CTLA-4 deficiency was lethal due to massive infiltration of tissues by polyclonally proliferating lymphocytes. CTLA-4 A49G polymorphism, which has been suggested to reduce the inhibitory function of the CTLA-4 molecule, was found to be associated with various autoimmune diseases in recent studies. Material and Methods: In this study, we evaluated the frequency of CTLA-4 A49G polymorphism in 46 patients with autoimmune hemolytic anemia (AIHA, 62 patients with immune thrombocytopenic purpura (ITP, and 150 healthy individuals. Results: Allele frequencies and genotype distributions were similar in both ITP and AIHA patients compared to healthy individuals. In subgroup analysis, however, we found that in chronic lymphocytic leukemia (CLL patients with AIHA (n=4, all patients had CTLA-4 A49G polymorphism (3 had AG, 1 had GG. There was no significant statistical association between G allele and systemic lupus erythematosus (SLE or AIHA.Conclusion: These data suggest that CTLA-4 A49G polymorphism does not contribute to the pathogenesis of lymphoproliferative diseases itself, nor does it increase the risk of autoimmune complications in patients with lymphoproliferative disease.

  8. Diminution of Oxidative Damage to Human Erythrocytes and Lymphocytes by Creatine: Possible Role of Creatine in Blood.

    Directory of Open Access Journals (Sweden)

    Neha Qasim

    Full Text Available Creatine (Cr is naturally produced in the body and stored in muscles where it is involved in energy generation. It is widely used, especially by athletes, as a staple supplement for improving physical performance. Recent reports have shown that Cr displays antioxidant activity which could explain its beneficial cellular effects. We have evaluated the ability of Cr to protect human erythrocytes and lymphocytes against oxidative damage. Erythrocytes were challenged with model oxidants, 2, 2'-azobis(2-amidinopropane dihydrochloride (AAPH and hydrogen peroxide (H2O2 in the presence and absence of Cr. Incubation of erythrocytes with oxidant alone increased hemolysis, methemoglobin levels, lipid peroxidation and protein carbonyl content. This was accompanied by decrease in glutathione levels. Antioxidant enzymes and antioxidant power of the cell were compromised while the activity of membrane bound enzyme was lowered. This suggests induction of oxidative stress in erythrocytes by AAPH and H2O2. However, Cr protected the erythrocytes by ameliorating the AAPH and H2O2 induced changes in these parameters. This protective effect was confirmed by electron microscopic analysis which showed that oxidant-induced cell damage was attenuated by Cr. No cellular alterations were induced by Cr alone even at 20 mM, the highest concentration used. Creatinine, a by-product of Cr metabolism, was also shown to exert protective effects, although it was slightly less effective than Cr. Human lymphocytes were similarly treated with H2O2 in absence and presence of different concentrations of Cr. Lymphocytes incubated with oxidant alone had alterations in various biochemical and antioxidant parameters including decrease in cell viability and induction of DNA damage. The presence of Cr attenuated all these H2O2-induced changes in lymphocytes. Thus, Cr can function as a blood antioxidant, protecting cells from oxidative damage, genotoxicity and can potentially increase their

  9. Homing receptor expression is deviated on CD56+ blood lymphocytes during pregnancy in Type 1 diabetic women.

    Directory of Open Access Journals (Sweden)

    Suzanne D Burke

    Full Text Available Type 1 Diabetes Mellitus (T1DM is characterized by an augmented pro-inflammatory immune state. This contributes to the increased risk for gestational complications observed in T1DM mothers. In normal pregnancies, critical immunological changes occur, including the massive recruitment of lymphocytes, particularly CD56bright NK cells, into early decidua basalis and a 2nd trimester shift towards Type 2 immunity. Decidual CD56bright NK cells arise at least partly from circulating progenitors expressing adhesion molecules SELL and ITGA4 and the chemokine receptors CXCR3 and CXCR4. In vitro studies show that T1DM reduces interactions between blood CD56+ NK cells and decidual endothelial cells by reducing SELL and ITGA4-based interactions. To address the mechanisms by which specific lymphocyte subsets may be recruited from the circulation during pregnancy and whether these mechanisms are altered in T1DM, flow cytometry was used to examine eight peripheral blood lymphocyte subsets (Type 1 (IL18R1+ and Type 2 (IL1RL1+ CD56bright NK, CD56dim NK, NKT and T cells from control and T1DM women. Blood was collected serially over pregnancy and postpartum, and lymphocytes were compared for expression of homing receptors SELL, ITGA4, CXCR3, and CXCR4. The decline of Type 1/Type 2 immune cells in normal pregnancy was driven by an increase in Type 2 cells that did not occur in T1DM. CD56bright NK cells from control women had the highest expression of all four receptors with greatest expression in 2nd trimester. At this time, these receptors were expressed at very low levels by CD56bright NK cells from TIDM patients. Type 1/Type 2 NKT cell ratios were not influenced by either pregnancy or TIDM. Our results suggest that T1DM alters immunological balances during pregnancy with its greatest impact on CD56bright NK cells. This implicates CD56bright NK cells in diabetic pregnancy complications.

  10. Scanning electron microscopy of interaction of peripheral blood lymphocytes from colonic cancer patients with human colonic cancer-derived cells; P-4788.

    Directory of Open Access Journals (Sweden)

    Sugihara,Mutsuto

    1979-12-01

    Full Text Available Peripheral blood lymphocytes and the various lymphocyte fractions from patients with cancer of the colon were cultivated with target cells (P-4788 derived from the colon cancer. Changes in the surface ultrastructure during tumor cell destruction were studied by scanning electron microscopy (SEM. P-4788 cells adhering to the coverslip showed various surface activity. The surfaces of some cells were relatively flat; others were smooth or had fine granules. Still other cells were villous, round or had marked blebs. When host lymphocytes were added to the target cells, adhesion of the two cell groups began by many fine projections. After incubation for 6 h, some lymphocytes had adhered to the target cells. Many lymphocytes had adhered to the target tumor cells by 24--48 h incubation. Ultimately the tumor cells became swollen and disrupted. Most lymphocytes adherent to the target cells had few microvilli. Lymphocytes after elimination of phagocytes by carbonyl iron treatment also adhered readily. Some target cells showed adhesion with lymphocytes passed through nylon-wool columns, although the number of lymphocytes adhering was fewer than in the case of lymphocytes not passed through nylon-wool columns. T cells were collected from lymphocytes that form rosettes with SRBC by isolation with NH4Cl. They had markedly elongated microvilli which in places were sparsely scattered and tended to be localized on the side, a finding which suggests loss of cell activity by the time of SEM. Only a few T cells adhered to target cells and they seemed to be T cells without activity. It was thought that there are cytotoxic cells among T cells and that the co-existence of T cells, non-T cells and monocytes caused target cell destruction.

  11. Natural background radiation induces cytogenetic radioadaptive response more effectively than occupational exposure in human peripheral blood lymphocytes

    Science.gov (United States)

    Monfared, A. Shabestani; Mozdarani, H.; Amiri, M.

    2003-01-01

    Ramsar, a city in the northern Iran, has the highest level of natural background radiation in the world. It has been clearly shown that low doses of ionising radiation can induce resistance to subsequent higher exposures. This phenomenon is termed radioadaptive response. We have compared induction of cytogenetic radioadaptive response by High Natural Background Radiation (HNBR) in Ramsar and X-ray occupational exposure as conditioning doses in human peripheral blood lymphocytes. 30 healthy control individuals, living in Ramsar but in normal background radiation areas, 15 healthy individuals from Talesh Mahalleh, a region with extraordinary high level of background radiation, and 7 X-ray radiographers working in Ramsar hospital located in normal natural background ionising radiation area were evaluated. Peripheral blood samples were prepared and exposed to challenge dose of 0 and 2 Gy. Lymphocytes were scored using analysis of metaphase, for the presence of chromosomal aberrations. An adaptive response was observed in HNBR and radiation workers groups in comparison with sham controls. A significant increase in adaptive response was observed in the HNBR group if compared with the occupationally exposed group. These findings indicate that both natural background radiation and occupational exposure could induce cytogenetic radioadaptive response and it is more significant regarding to natural background ionising radiation.

  12. Strong association between long and heterogeneous telomere length in blood lymphocytes and bladder cancer risk in Egyptian.

    Science.gov (United States)

    Wang, Hongkun; Wang, Ying; Kota, Krishna K; Kallakury, Bhaskar; Mikhail, Nabiel N; Sayed, Douaa; Mokhtar, Ahmed; Maximous, Doaa; Yassin, Etemad H; Gouda, Iman; Sobitan, Adebiyi; Sun, Bing; Loffredo, Christopher A; Zheng, Yun-Ling

    2015-11-01

    Although it is widely recognized that telomere dysfunction plays an important role in cancer, the relationship between telomere function and bladder cancer risk is not well defined. In a case-control study of bladder cancer in Egypt, we examined relationships between two telomere features and bladder cancer risk. Telomere fluorescent in situ hybridization was used to measure telomere features using short-term cultured blood lymphocytes. Logistic regression was used to estimate the strength of association between telomere features and the risk of urothelial carcinoma of the bladder. High telomere length variation (TLV) across all chromosomal ends was significantly associated with an increased risk of bladder cancer [adjusted odds ratios (OR) = 2.22, 95% confidence interval (CI) = 1.48-3.35], as was long average telomere length (OR = 3.19, 95% CI = 2.07, 4.91). Further, TLV and average telomere length jointly affected bladder cancer risk: when comparing individuals with long telomere length and high TLV to those with short telomere length and low TLV, the adjusted OR was 14.68 (95% CI: 6.74-31.98). These associations were stronger among individuals who are 60 years of age or younger. In summary, long and heterogeneous telomere length in blood lymphocytes was strongly associated with an increased bladder cancer risk in Egyptian and the association was modulated by age.

  13. Thymus cells in myasthenia gravis selectively enhance production of anti-acetylcholine-receptor antibody by autologous blood lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Newsom-Davis, J.; Willcox, N.; Calder, L.

    1981-11-26

    We investigated the role of the thymus in 16 patients with myasthenia gravis without thymoma by studying the production of anti-acetylcholine-receptor antibody by thymic and blood lymphocytes cultured alone or together. In 10 responders (with the highest receptor-antibody titers in their plasma), cultured thymic cells spontaneously produced measurable receptor antibody. Receptor-antibody production by autologous blood lymphocytes was enhanced by the addition of responder's thymic cells, irradiated to abrogate antibody production and suppression (P<0.01). This enhancement was greater and more consistent than that by pokeweed mitogen; it depended on viable thymic cells, appeared to be selective for receptor antibody, and correlated with the ratio of thymic helper (OKT4-positive or OKT4+) to suppressor (OKT8+) T cells (P<0.01). These results suggest that myasthenic thymus contains cell-bound acetylcholine-receptor-like material or specific T cells (or both) that can aid receptor-antibody production. This may be relevant to the benefits of thymectomy in myasthenia and to the breakdown in self-tolerance in this and other autoimmune diseases.

  14. Comparison of differences between dicentric assay and translocation analysis for biodosimetry in cultured peripheral blood lymphocytes of Korean individuals

    Energy Technology Data Exchange (ETDEWEB)

    Park, Hyun Jin; Park, Mi Young; Seo, Min Ji; Kwon, Hee Kyung; Lee, Su Jae; Lee, Yun Sil; Ji, Young Hoon; Choi, Soo Yong; Cho, Chul Koo; Kim, Tae Hwan; Kang, Chang Mo [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

    2004-07-01

    Chromosome aberrations are considered to be important indicators of induced DNA damage and genomic instability. For this reason, they constitute the main parameter used to monitor individuals exposed to radiation. Biological dosimetry using the analysis of dicentrics in human lymphocytes is well established, especially in case of acute exposure, when the blood samples are taken within a few weeks. However, dicentric analysis is not an adequate parameter in case of chronic exposure, because these aberrations are unstable with time, and have a limited use for dose assessment of past exposures. In contrast to dicentrics, however, translocations are considered stable in cell division and so the yield should not fall with time. In the present study, using FISH-chromosome painting analysis with the dose-response curve for chromosome aberrations, we monitored the stable and unstable chromosome aberrations of 2 Korean's periperal blood lymphocytes irradiated in vitro with {gamma}-rays from {sup 137}Cs (doses between 0.0 and 2.0 Gy). By using the dose-response curve for chromosome aberration, our aim was to estimate the absorbed doses, and then establish comparison with the results obtained by conventional dicentric analysis, thus taking the opportunity to test the validity of chromosome aberration analysis by FISH painting method for retrospective biodosimetry in Korean individual.

  15. Increased percentage of CD8 CD28– suppressor lymphocytes in peripheral blood and skin infiltrates correlates with advanced disease in patients with cutaneous T-cell lymphomas

    Directory of Open Access Journals (Sweden)

    Donata Urbaniak-Kujda

    2009-07-01

    Full Text Available Introduction: T cells with the CD8 CD28– phenotype are CD8 lymphocytes with regulatory function. Their increased numbers were observed in infections, autoimmune and neoplastic diseases, and in elderly healthy individuals. CD8 CD28– lymphocyte levels in patients with cutaneous T-cell lymphoma (CTCL has not yet been described. The aim of the study was to determine their levels in these patients’ peripheral blood and cutaneous infiltrates and their relation to the clinical stage of disease.Material/Methods: Forty-one untreated patients, 26 males and 15 females, with CTCL were enrolled in the study. CD8 CD28– lymphocyte levels were determined by flow cytometry in peripheral blood and by immunochemistry in skin infiltrates.Results: The percentage of CD8 CD28– lymphocytes in the peripheral blood of the patients was significantly higher than in the controls. Patients with advanced disease displayed a higher percentage of CD8 CD28– lymphocytes in the peripheral blood and skin than did the individuals with early stages of the disease. Moreover, positive correlations between CD8 CD28– lymphocyte level in peripheral blood and age, clinical stage, and the levels in the skin infiltrates was revealed. Additionally, the percentage of CD8 CD28– T cells in the skin infiltrates correlated positively with age and clinical stage of the disease.Conclusions: These data suggest that CD8 CD28– lymphocytes play an important role in the development of immunotolerance in the progression of cutaneous T-cell lymphoma.

  16. Effects of doxycycline on haematology, blood chemistry and peripheral blood lymphocyte subsets of healthy dogs and dogs naturally infected with Ehrlichia canis.

    Science.gov (United States)

    Villaescusa, A; García-Sancho, M; Rodríguez-Franco, F; Tesouro, M Á; Sainz, Á

    2015-06-01

    Canine monocytic ehrlichiosis (CME), caused by Ehrlichia canis, is a vector-borne disease with a worldwide distribution. It has been proposed that the pathogenesis, clinical severity and outcome of disease caused by Ehrlichia spp. can be attributed to the immune response rather than to any direct rickettsial effect. Moreover, doxycycline, the antimicrobial of choice for the treatment of CME, has immunomodulatory and anti-inflammatory properties associated with blood leukocyte proliferation function, cytokine synthesis, and matrix metalloproteinase activity. In order to assess the potential effects of doxycycline, dependent and independent of its antimicrobial activity, the present study compared changes in haematology, blood chemistry and circulating lymphocyte subpopulations in 12 healthy dogs and 20 dogs with CME after doxycycline therapy. Some changes were recorded only in the CME affected dogs, probably due to the antimicrobial effect of doxycycline. However, increases in mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, platelet count and α2-globulins, and decreased plasma creatinine were observed in both healthy and CME affected dogs. The absolute count of B lymphocytes (CD21(+)) increased initially, but then decreased until the end of the study period in both groups. A potential effect of doxycycline unrelated to its antimicrobial activity against E. canis is suggested, taking into account the results observed both in healthy dogs and in dogs with CME.

  17. SILVER NANOPARTICLES AND EXPERESSION OF MOLECULAR MARKERS IN LYMPHOCYTE ACTIVATION AND MARKER OF AUTOIMMUNE PROCESSES IN PERIPHERAL BLOOD OF PATIENTS WITH VIRAL CORNEAL PATHOLOGY

    Directory of Open Access Journals (Sweden)

    Ulyanov V.A.

    2015-08-01

    Full Text Available The influence of the nanoparticles of silver on the expression of molecular markers activation of lymphoid cells CD7+, CD25+, CD38+, CD45+, CD54+, CD95+, CD150+ and CD5+ – marker of the autoimmune process, as well as on phagocytic activity of neutrophils in patients with viral pathologies of the cornea was studied in vitro. In the Laboratory of Immunology, SI Institute of Eye Diseases and Tissue Therapy NAMS of Ukraine was developed technique of cultivation of peripheral blood lymphocytes with immunomodulation drugs, followed by determination of changes in the level of expression of molecular markers of lymphocyte activation. Assessment of the level of expression of molecular markers of activation of peripheral blood lymphocytes was performed method using a panel of monoclonal antibodies, CD5+, CD7+, CD25+, CD38+, CD45+, CD54+, CD95+ and CD 150+. The study was conducted in vitro with the peripheral lymphocytes the blood of 23 patients of viral pathology of the cornea. Our studies of the effects of nanosilver particles in vitro on the state of expression of molecular markers of activation of peripheral blood lymphocytes and phagocytic activity of neutrophils in patients with viral corneal pathology, showed a significant increase in the level of expression of the CD7+, CD25+, CD45+ and phagocytic activity of neutrophils after application silver nanoparticles.

  18. THE ROLE OF PROTEIN OXIDATIVE MODIFICATION IN REDOX-REGULATION OF CASPASE-3 ACTIVITY IN BLOOD LYMPHOCYTES DURING OXIDATIVE STRESS IN VITRO

    Directory of Open Access Journals (Sweden)

    O. L. Nosareva

    2015-01-01

    Full Text Available The formation of oxidative stress lies at the heart of many frequent and socially-important diseases. Blood lymphocytes are the cells which provide immunological control of our organism. As a result of their function implementation blood lymphocytes contact with different endogenic and exogenic factors, which can lead to active oxygen species production activation, macromolecules oxidative modification and to cell survival alteration. At the present time it is essential to expand and deepen the fundamental knowledge of blood lymphocytes apoptosis regulation peculiarities. The research objective was to establish the interaction among alterations of glutathione system condition, carbonylation level, protein glutathionylation and caspase-3 activity in blood lymphocytes during oxidative stress in vitro.Material and Methods. The material for research was blood lymphocytes cultivated with addition of hydrogen peroxide in final concentration of 0,5 mmol and/or protein SH-group inhibitor N-ethylmaleimide – 5 mmol, protector – 5 mmol – 1,4-dithioerythritol. Reduced, oxidized and protein-bound glutathione concentration was measured by method of spectropho-tometry, additionally, the ratio size of reduced to oxidized thiol fraction was estimated. With help of enzymoimmunoassay the level of protein carbonyl derivatives was evaluated; caspase-3 activity was registered by spectrofluorometric method.Results. Protein SH-group blocking in blood lymphocytes during oxidative stress in vitro was accompanied by protein-bound glutathione concentration rapid decrease in connection with increase of protein carbonyl derivatives content and caspase-3 activity. Protein SH-group protection in blood lymphocytes during oxidative stress in vitro was accompanied by concentration increase of protein-bound glutathione and protein carbonyl derivatives under comparable values of enzyme activity under study.Conclusion. The carried out research shows that caspase-3 and protein

  19. The in vitro genotoxic effects of a commercial formulation of alpha-cypermethrin in human peripheral blood lymphocytes.

    Science.gov (United States)

    Kocaman, Ayşe Yavuz; Topaktaş, Mehmet

    2009-01-01

    alpha-Cypermethrin, a highly active pyrethroid insecticide, is effective against a wide range of insects encountered in agriculture and animal husbandry. The potential genotoxicity of a commercial formulation of alpha-cypermethrin (Fastac 100 EC, containing 10% alpha-cypermethrin as the active ingredient) on human peripheral lymphocytes was examined in vitro by sister chromatid exchange (SCE), chromosomal aberrations (CAs), and micronucleus (MN) tests. The human lymphocytes were treated with 5, 10, 15, and 20 microg/ml of alpha-cypermethrin for 24- and 48-hr. alpha-Cypermethrin induced SCEs and CAs significantly at all concentrations and treatment times and MN formation was significantly induced at 5 and 10 microg/ml of alpha-cypermethrin when compared with both the control and solvent control. Binuclear cells could not be detected sufficiently in the highest two concentration of alpha-cypermethrin (15 and 20 microg/ml) for both the 24- and 48-hr treatment times. alpha-Cypermethrin decreased the proliferation index (PI) at three high concentrations (10, 15, and 20 microg/ml) for both treatment periods as compared with the control groups. In addition, alpha-cypermethrin reduced both the mitotic index (MI) and nuclear division index (NDI) significantly at all concentrations for two treatment periods. The PI and MI were reduced by alpha-cypermethrin in a concentration-dependent manner during both treatment times. In general, alpha-cypermethrin showed higher cytotoxic and cytostatic effects than positive control (MMC) at the two highest concentrations for the 24- and 48-hr treatment periods. The present study is the first to report the genotoxic and cytotoxic effects of commercial formulation of alpha-cypermethrin in peripheral blood lymphocytes.

  20. The Induction of Chromosome Aberrations and Micronuclei in Human Peripheral Blood Lymphocytes at Low Doses of Radiation

    CERN Document Server

    Shmakova, N L; Krasavin, E A; Melnikova, L A; Fadeeva, T A

    2003-01-01

    The chromosome damage induced by the low doses of gamma-irradiation with ^{60}Co and X-rays in peripheral blood lymphocytes has been studied using different cytogenetic assays. Isolated lymphocytes were exposed to 0.01-1.0 Gy, simulated by PHA, and analysed for chromosome aberrations by the metaphase and the anaphase methods, by the micronucleus assay. Despite the quantitative differences in the amount of chromosome damage revealed by different methods, all of them demonstrated complex nonlinear dose dependence of the frequency of aberrant cells and aberrations. At the dose range of 0.01-0.05 Gy the cells showed the highest radiosensitivity; at 0.05-0.5 Gy the dose-independent induction of chromosome damage was revealed. At the doses of 0.5-1.0 Gy the dose-effect curves became linear with the decreased slope compared with the initial one (by a factor of 5 to 10 for different criteria) reflecting a higher radioresistance of the cells. These data confirm the idea that the direct linear extrapolation of high-dos...

  1. Dioxidine-induced changes in genome-wide DNA methylation in a culture of peripheral blood lymphocytes.

    Science.gov (United States)

    Smirnikhina, S A; Voronina, E S; Lavrov, A V; Bochkov, N P

    2013-06-01

    We studied the effect of dioxidine on genome-wide methylation in short-term cultures of peripheral blood lymphocytes derived from healthy donors. Methylation was evaluated in lymphocytes before culturing, after 25 h in culture, and 1 h after addition of dioxidine in two concentrations (0.1 and 0.01 mg/ml). The total time in culture was 25 h. The level of methylation was assessed using methyl-sensitive single-cell gel electrophoresis ("comet assay") with additional restriction with HpaII amd MspI. Significant individual differences were found in the levels of methylation in both native cells and in cells treated with dioxidine in both concentrations. Mean group indicators of methylation did not differ before culturing and after 25 h in culture (45.28 and 44.80%, respectively). The mean group rate of methylation increased to 46.14% (p<0.001) after dioxidine treatment in a concentration of 0.01 mg/ml. Dioxidine in 0.1 mg/ml reduced the level of methylation (mean group rate 42.31%; p<0.001).

  2. The Changes of Protein Kinase C Activity in Peripheral Blood Lymphocytes in the Patients with Obstructive Jaundice and the Implication

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The roles of protein kinase C (PKC) signal pathway in the pathogenesis of obstructive jaundice were studied. PKC from cytosolic and membrane fractions of peripheral blood lymphocytes (PBL) in 51 patients with obstructive jaundice and 16 cases of normal controls was isolated and purified. The activities of PKC were determined by radioactive isotope γ-32P-ATP-catalyzing assay. The results showed that the total PKC activities in PBL in the patients with obstructive jaundice were significantly increased as compared with those in the normal controls (P<0.01). Moreover, the membrane PKC activities and their percentages of the total PKC activities were higher in obstructive jaundice group than in those in the normal controls (P<0.05). The total PKC activities in PBL in the patients with obstructive jaundice were significantly positively correlated with the levels of soluble IL-2 receptor (sIL-2R) (r=0.58, P<0.01) and the degree of jaundice (T-BIL) (r=0.67, P<0.01) in serum. It was concluded that the activities of PKC signal pathway was related with the degree of T-BIL. PKC signal pathway might took part in the activation of T-lymphocytes in the patients with obstructive jaundice and play an important role in the immune regulation and the assessment of pathosis in the patients with obstructive jaundice.

  3. Analysis of the CDR3 Length Repertoire and the Diversity of TCRα Chain in Human Peripheral Blood T Lymphocytes

    Institute of Scientific and Technical Information of China (English)

    Xinsheng Yao; Ying Diao; Wanbang Sun; Junmin Luo; Ming Qin; Xianying Tang

    2007-01-01

    Analysis of complementarity determining region 3 (CDR3) length of T lymphocyte receptors (TCRs) by immunoscope spectratyping technique has been used successfully to investigate the diversity of TCR in autoimmune diseases and infection diseases. In this study, we investigated the patterns of CDR3 length distribution for all 32 TCR AV gene families in human peripheral blood lymphocytes of four normal volunteers by the immunoscope spectratyping technique. It was found that PCR products exhibited an obscure band on 1.5% agarose gel electrophoresis. Each TCR AV family exhibited more than 8 bands on 6% sequencing gel electrophoresis. The CDR3 spectratyping of all TCR AV families showed a standard Gaussian distribution with different CDR3 length,and the expression frequency of CDR3 was similar among the gene families. Most of CDR3 in TCR AV family recombine in frame. However, some of the CDR3 showed out-of frame gene rearrangement. Additionally, we found that in some of TCR AV families there were 18 amino acid discrepancies between the longest CDR3 and shortest CDR3. These results may be helpful to further study the recombination mechanism of human TCR genes, the TCR CDR3 gene repertoire, and the repertoire drift in health people and disease state.

  4. Use of whole blood lymphocyte stimulation test for immunocompetency studies in bald eagles, red-tailed hawks, and great horned owls.

    Science.gov (United States)

    Redig, P T; Dunnette, J L; Sivanandan, V

    1984-11-01

    Mitogen-induced whole blood lymphocyte stimulation tests for immunocompetency studies in bald eagles (Haliaeetus leucocephalus), red-tailed hawks (Buteo jamaicensis), and great horned owls (Bubo virginianus) were developed. Combinations of incubation times, blood dilutions, concentrations of [3H]thymidine and [125I]2-deoxyuridine, antibiotics, phytohemagglutinin-P, and concanavalin A were tested for their effects on the stimulation index (SI). An antibiotic combination of gentamicin plus amphotericin B yielded low SI with lymphocytes from bald eagles, but not with lymphocytes from great horned owls or red-tailed hawks. Penicillin plus streptomycin caused no such depression of SI. Lymphocytes from all 3 species yielded maximum responses with a 48-hour prelabel and 12- to- 16 hour postlabel incubation period at 41 C and 1:20 blood dilution. Optimal mitogen concentrations for lymphocytes from bald eagles, red-tailed hawks, and great horned owls were 25 micrograms, 10 micrograms, and 10 micrograms of phytohemagglutinin-P/well, respectively, and 2.5 micrograms, 10 micrograms, and 10 micrograms of concanavalin A/well, respectively. Differences in SI were not seen between the 2 radioactive labels. The optimal concentration of the [3H]thymidine label ranged from 0.06 to 0.125 microCi/well.

  5. Preoperative Monocyte-to-Lymphocyte Ratio in Peripheral Blood Predicts Stages, Metastasis, and Histological Grades in Patients with Ovarian Cancer

    Directory of Open Access Journals (Sweden)

    Jiangdong Xiang

    2017-02-01

    Full Text Available PURPOSE: The monocyte-to-lymphocyte ratio (MLR has been shown to be associated with the prognosis of various solid tumors. This study sought to evaluate the important value of the MLR in ovarian cancer patients. METHODS: A total of 133 ovarian cancer patients and 43 normal controls were retrospectively reviewed. The patients' demographics were analyzed along with clinical and pathologic data. The counts of peripheral neutrophils, lymphocytes, monocytes, and platelets were collected and used to calculate the MLR, neutrophil-to-lymphocyte ratio (NLR. and platelet-to-lymphocyte ratio (PLR. The optimal cutoff value of the MLR was determined by using receiver operating characteristic curve analysis. We compared the MLR, NLR, and PLR between ovarian cancer and normal control patients and among patients with different stages and different grades, as well as between patients with lymph node metastasis and non–lymph node metastasis. We then investigated the value of the MLR in predicting the stage, grade, and lymph node positivity by using logistic regression. The impact of the MLR on overall survival (OS was calculated by Kaplan-Meier method and compared by log-rank test. RESULTS: Statistically significant differences in the MLR were observed between ovarian cancer patients and normal controls. However, no difference was found for the NLR and PLR. Highly significant differences in the MLR were found among patients with different stages (stage I-II and stage III-IV, grades (G1 and >G1, and lymph node metastasis status. The MLR was a significant and independent risk factor for lymph node metastasis, as determined by logistic regression. The optimal cutoff value of the MLR was 0.23. We also classified the data according to tumor markers (CA125, CA199, HE4, AFP, and CEA and conventional coagulation parameters (International Normalized Ratio [INR] and fibrinogen. Highly significant differences in CA125, CA199, HE4, INR, fibrinogen levels, and lactate

  6. In vitro antigenic stimulation of peripheral blood and lymph node lymphocytes of sensitized guinea-pigs: the effect of a second administration of antigen in vivo

    Science.gov (United States)

    Housley, J.; Gell, P. G. H.

    1969-01-01

    Stimulation of DNA synthesis by a guinea-pig albumin—orthanilic acid conjugate (AO) and by tuberculin purified protein derivative (PPD) was obtained in in vitro cultures of peripheral blood and lymph node lymphocytes from guinea-pigs with delayed hypersensitivity to these antigens. Animals sensitized to both AO and PPD were given a further injection of 5 μg AO, intravenously, 8 hours before killing for in vitro studies. In these guinea-pigs, peripheral blood cultures, but not lymph node cultures, showed greater DNA synthesis in response to both AO and PPD than cultures from controls not given a further injection of AO. It is suggested that the further increase in DNA synthesis was due to non-specific lymphocyte `activation' following the interaction of antigen and specifically sensitized lymphocytes. PMID:5352364

  7. Aging of immune system: Immune signature from peripheral blood lymphocyte subsets in 1068 healthy adults.

    Science.gov (United States)

    Qin, Ling; Jing, Xie; Qiu, Zhifeng; Cao, Wei; Jiao, Yang; Routy, Jean-Pierre; Li, Taisheng

    2016-05-01

    Aging is a major risk factor for several conditions including neurodegenerative, cardiovascular diseases and cancer. Functional impairments in cellular pathways controlling genomic stability, and immune control have been identified. Biomarker of immune senescence is needed to improve vaccine response and to develop therapy to improve immune control. To identify phenotypic signature of circulating immune cells with aging, we enrolled 1068 Chinese healthy volunteers ranging from 18 to 80 years old. The decreased naïve CD4+ and CD8+ T cells, increased memory CD4+ or CD8+ T cells, loss of CD28 expression on T cells and reverse trend of CD38 and HLA-DR, were significant for aging of immune system. Conversely, the absolute counts and percentage of NK cells and CD19+B cells maintained stable in aging individuals. The Chinese reference ranges of absolute counts and percentage of peripheral lymphocyte in this study might be useful for future clinical evaluation.

  8. Expressions of HSP 70 and NF-κB in the peripheral blood lymphocyte of chronic gastritis patients of different syndrome patterns

    Institute of Scientific and Technical Information of China (English)

    胡玲

    2012-01-01

    Objective To study the expressions of heat shock protein 70 (HSP 70) and nuclear factor-kappa B(NF-κB) in the peripheral blood lymphocyte of chronic gastritis (CG) patients of Pi-Wei hygropyrexia syndrome (PWHS) and Pi-qi deficiency syndrome(PQDS),and to explore their correlation with Helicobacter pylori(Hp) infection. Methods

  9. T-Lymphocytes Traffic into the Brain across the Blood-CSF Barrier: Evidence Using a Reconstituted Choroid Plexus Epithelium.

    Science.gov (United States)

    Strazielle, Nathalie; Creidy, Rita; Malcus, Christophe; Boucraut, José; Ghersi-Egea, Jean-François

    2016-01-01

    An emerging concept of normal brain immune surveillance proposes that recently and moderately activated central memory T lymphocytes enter the central nervous system (CNS) directly into the cerebrospinal fluid (CSF) via the choroid plexus. Within the CSF space, T cells inspect the CNS environment for cognate antigens. This gate of entry into the CNS could also prevail at the initial stage of neuroinflammatory processes. To actually demonstrate T cell migration across the choroidal epithelium forming the blood-CSF barrier, an in vitro model of the rat blood-CSF barrier was established in an "inverse" configuration that enables cell transmigration studies in the basolateral to apical, i.e. blood/stroma to CSF direction. Structural barrier features were evaluated by immunocytochemical analysis of tight junction proteins, functional barrier properties were assessed by measuring the monolayer permeability to sucrose and the active efflux transport of organic anions. The migratory behaviour of activated T cells across the choroidal epithelium was analysed in the presence and absence of chemokines. The migration pathway was examined by confocal microscopy. The inverse rat BCSFB model reproduces the continuous distribution of tight junction proteins at cell margins, the restricted paracellular permeability, and polarized active transport mechanisms, which all contribute to the barrier phenotype in vivo. Using this model, we present experimental evidence of T cell migration across the choroidal epithelium. Cell migration appears to occur via a paracellular route without disrupting the restrictive barrier properties of the epithelial interface. Apical chemokine addition strongly stimulates T cell migration across the choroidal epithelium. The present data provide evidence for the controlled migration of T cells across the blood-CSF barrier into brain. They further indicate that this recruitment route is sensitive to CSF-borne chemokines, extending the relevance of this

  10. Human Prolactin Improves Engraftment and Reconstitution of Human Peripheral Blood Lymphocytes in SCID Mice

    Institute of Scientific and Technical Information of China (English)

    Rui Sun; Jian Zhang; Cai Zhang; Jianhua Zhang; Shujuan Liang; Anyuan Sun; Junfu Wang; Zhigang Tian

    2004-01-01

    Recombinant human prolactin (rhPRL) was administered to huPBL-SCID mice to determine its effects on human immunologic reconsfitution and function. The huPBL-SCID mice were given 10 μg I.p. Injection of rhPRL every other day for a total of 10 injections after huPBL were transferred. The results demonstrated that rhPRL improved the engraftment of lymphocytes into thymus, lymph nodes and spleens, showing that the cellularities of these organs increased although the cellularities tended to vary depending on the donor. The amounts of human T cells (HLA-ABC+/CD3+) increased greatly in thymus (14.2 folds), spleen (4.16 folds) and lymph nodes (40.18 folds) after rhPRL injections. The amounts of human B cells (HLA-ABC+/CD19+) also increased greatly in lymph nodes (42.5 folds) and spleen (5.78 folds). The lymph node cells from the rhPRL-treated huPBL-SCID mice were more sensitive to PHA stimulation ([3H] thymidine incorporation). The supernatant of PHA-stimulated PBL from rhPRL-treated huPBL/SCID chimerism contained more cytokines (IFN-γ and IL-2). The natural cytotoxicity against human sensitive target cells, K562 cells, from spleen and bone marrow of hPBL/SCID chimerism was significantly enhanced by rhPRL administration. The lymph node cells were stimulated with LPS in vitro for 3 days and the lymphocytes from the rhPRL-treated huPBL-SCID mice were more sensitive to mitogen stimulation. Both serum total IgG level and IgM level of rhPRL-treated huPBL/SCID chimerism were increased, and even without DT-rechallenge the base line of DT-specific IgG was elevated after rhPRL treatment in huPBL-SCID mice. Thus, rhPRL stimulation promotes reconstitution of human immune system in huPBL-SCID mice.

  11. Human Prolactin Improves Engraftment and Reconstitution of Human Peripheral Blood Lymphocytes in SCID Mice

    Institute of Scientific and Technical Information of China (English)

    RuiSun; JianZhang; CaiZhang; JianhuaZhang; ShujuanLiang; AnyuanSun; JunfuWang; ZhigangTian

    2004-01-01

    Recombinant human prolactin (rhPRL) was administered to huPBL-SCID mice to determine its effects on human immunologic reconstitution and function. The huPBL-SCID mice were given 10 μg i.p. injection of rhPRL every other day for a total of 10 injections after huPBL were transfered. The results demonstrated that rhPRL improved the engraftment of lymphocytes into thymus, lymph nodes and spleens, showing that the cellularities of these organs increased although the cellularities tended to vary depending on the donor. The amounts of human T cells (HLA-ABC+/CD3+) increased greatly in thymus (14.2 folds), spleen (4.16 folds) and lymph nodes (40.18 folds) after rhPRL injections. The amounts of human B cells (HLA-ABC+/CD19+) also increased greatly in lymph nodes (42.5 folds) and spleen (5.78 folds). The lymph node cells from the rhPRL-treated huPBL-SCID mice were more sensitive to PHA stimulation (〔3H〕thymidine incorporation). The supernatant of PHA-stimulated PBL from rhPRL-treated huPBL/SCID chimerism contained more cytokines (IFN-γ and IL-2). The natural cytotoxicity against human sensitive target cells, K562 cells, from spleen and bone marrow of hPBL/SCID chimerism was significantly enhanced by rhPRL administration. The lymph node cells were stimulated with LPS in vitro for 3 days and the lymphocytes from the rhPRL-treated huPBL-SCID mice were more sensitive to mitogen stimulation. Both serum total IgG level and IgM level of rhPRL-treated huPBL/SCID chimerism were increased, and even without DT-rechallenge the base line of DT-specific IgG was elevated after rhPRL treatment in huPBL-SCID mice. Thus, rhPRL stimulation promotes reconstitution of human immune system in huPBL-SCID mice. Cellular & Molecular Immunology. 2004;1(2):129-136.

  12. Haploinsufficiency for BRCA1 is associated with normal levels of DNA nucleotide excision repair in breast tissue and blood lymphocytes

    Directory of Open Access Journals (Sweden)

    Johnson Jennifer M

    2005-06-01

    Full Text Available Abstract Background Screening mammography has had a positive impact on breast cancer mortality but cannot detect all breast tumors. In a small study, we confirmed that low power magnetic resonance imaging (MRI could identify mammographically undetectable tumors by applying it to a high risk population. Tumors detected by this new technology could have unique etiologies and/or presentations, and may represent an increasing proportion of clinical practice as new screening methods are validated and applied. A very important aspect of this etiology is genomic instability, which is associated with the loss of activity of the breast cancer-predisposing genes BRCA1 and BRCA2. In sporadic breast cancer, however, there is evidence for the involvement of a different pathway of DNA repair, nucleotide excision repair (NER, which remediates lesions that cause a distortion of the DNA helix, including DNA cross-links. Case presentation We describe a breast cancer patient with a mammographically undetectable stage I tumor identified in our MRI screening study. She was originally considered to be at high risk due to the familial occurrence of breast and other types of cancer, and after diagnosis was confirmed as a carrier of a Q1200X mutation in the BRCA1 gene. In vitro analysis of her normal breast tissue showed no differences in growth rate or differentiation potential from disease-free controls. Analysis of cultured blood lymphocyte and breast epithelial cell samples with the unscheduled DNA synthesis (UDS assay revealed no deficiency in NER. Conclusion As new breast cancer screening methods become available and cost effective, patients such as this one will constitute an increasing proportion of the incident population, so it is important to determine whether they differ from current patients in any clinically important ways. Despite her status as a BRCA1 mutation carrier, and her mammographically dense breast tissue, we did not find increased cell

  13. Micronucleus frequency is increased in peripheral blood lymphocytes of nuclear power plant workers.

    Science.gov (United States)

    Hadjidekova, Valeria B; Bulanova, Minka; Bonassi, Stefano; Neri, Monica

    2003-12-01

    Nuclear power plant workers are exposed to ionizing radiation at relatively low doses and for prolonged periods of time. To investigate the extent of genetic damage in these workers, a group of 133 nuclear power plant workers and 39 healthy controls were compared using the cytokinesis-block micronucleus assay. The frequency of micronuclei was significantly increased in peripheral lymphocytes of nuclear power plant workers (20.5 +/- 9.7% compared to 13.7 +/- 5.9%). A significant dose-response relationship was observed between micronucleus (MN) frequency and both the accumulated dose and the duration of employment (P < 0.01 for both variables after adjusting for age, gender and cigarette smoking) with an evident leveling off for exposures over 200 mSv. Accumulated dose and duration of employment were significantly correlated but exerted independent effects on MN frequency. For non-occupational parameters, age was significantly associated with the frequency of micronuclei, while gender was not. Smoking habit showed no overall effect, whereas increased chromosome damage was evident in smokers of more than 20 cigarettes per day. In conclusion, a dose-related association between MN frequency and exposure to ionizing radiation was evident in nuclear power plant workers, encouraging the application of the cytokinesis-block micronucleus assay in biomonitoring studies of human populations with prolonged exposure to ionizing radiation.

  14. Classical scrapie prions in ovine blood are associated with B lymphocytes and platelets-rich plasma

    Science.gov (United States)

    Classical scrapie is a naturally occurring fatal brain disease of sheep and goats which is caused by prions, a novel class of infectious agent, and is accompanied by the accumulation of abnormal isoforms of prion protein (PrP-Sc) in certain neural and lymphoid tissues. Although collection of a blood...

  15. Hormonal differences in peripheral blood and gene profiling in the liver and lymphocytes in Japanese black cattle with growth retardation.

    Science.gov (United States)

    Ishida, Sakura; Yonezawa, Tomohiro; Eirai, Sayoko; Kizaki, Keiichiro; Hashizume, Kazuyoshi; Tomioka, Michiko; Kurose, Yohei; Hirano, Takashi; Watanabe, Daisaku

    2013-01-31

    Japanese Black cattle occasionally demonstrate growth retardation despite sufficient nutrient intake. To clarify hormonal and transcriptional characteristics, we investigated differences in blood components, including hormones, and differences in exhaustive gene expressions in the liver and peripheral lymphocytes of six cattle with growth retardation (GR cattle) and eight control cattle of the same age and pedigree with normal growth. Hematocrit values and concentrations of hemoglobin, serum albumin, total cholesterol, insulin-like growth factor 1 (IGF-1), thyroxine and insulin in GR cattle were significantly lower than those in controls. GR cattle also excreted higher levels of GH. We used three GR and three control cattle for a microarray analysis in the liver and found that 279 gene expressions were significantly different. However, gene expressions related to the GH-IGF-1 axis, such as the GH receptor and IGF-1, were not significantly different from those of controls. Immune-related gene expressions were significantly lower. To clarify these gene expression levels, peripheral lymphocytes were used for real-time RT-PCR. The expression rates of genes that were significantly lower in the liver, such as chemokine ligand 8, interferon gamma receptor 1 and immunoglobulin light chain VJ region were also significantly lower in three GR cattle than those in the three control cattle. These results suggest that the cause of growth retardation in the present study was due to other factors, not abnormal gene expressions of factors related to the GH-IGF-1 axis in the liver, and that GR cattle were susceptible to infectious disease.

  16. Genetic and oxidative damage of peripheral blood lymphocytes in workers with occupational exposure to coal.

    Science.gov (United States)

    Rohr, Paula; Kvitko, Kátia; da Silva, Fernanda R; Menezes, Ana Paula Simões; Porto, Carem; Sarmento, Merielen; Decker, Natália; Reyes, Juliana M; Allgayer, Mariangela da C; Furtado, Tatiane Chao; Salvador, Mirian; Branco, Catia; da Silva, Juliana

    2013-12-12

    Coal is an important fossil fuel used to generate energy. Coal dust is constituted primarily of hydrocarbons and metals. During coal extraction, large quantities of coal dust particles are emitted, contributing to environmental pollution. Coal miners are constantly exposed to coal dust and its derivatives. The goal of this study was to evaluate the potential genotoxic effects of coal and oxidative stress in individuals from Candiota who were exposed to coal as part of their occupation. The comet assay and micronucleus (MN) test were used to assess these effects. This study involved 128 male participants of whom 71 reported work that included exposure to coal (exposed group) and 57 reported working at different jobs (unexposed group). The exposed group had a significantly increased damage index and damage frequency, as assessed using the comet assay, and increased MN and nucleoplasmic bridge frequencies, as assessed using the MN assay, compared with unexposed individuals. Significant and positive correlations between MN frequencies in the lymphocytes and buccal cells of control and exposed individuals were observed. The exposed individuals presented lower average levels of thiobarbituric acid reactive substances (TBARS) and catalase activity (CAT), while the mean superoxide dismutase activity (SOD) levels were higher in this group. The exposed group also had higher hematocrit levels. No correlation between DNA damage and inorganic elements, as identified using PIXE, was found; however, there was a correlation between the damage index and zinc. The evidence that exposure to coal and its derivatives presents a genetic hazard demonstrates the need for protective measures and educational programs for coal miners.

  17. Analytic errors in Sysmex-generated hematology results in blood from a dog with chronic lymphocytic leukemia.

    Science.gov (United States)

    Novacco, Marilisa; Martini, Valeria; Grande, Carmen; Comazzi, Stefano

    2015-09-01

    A blood sample from a 14-year-old dog was submitted to the veterinary diagnostic laboratory of the University of Milan for marked leukocytosis with atypical cells. A diagnosis of chronic T-cell lymphocytic leukemia (CLL) was made based on blood smear evaluation and flow cytometric phenotyping. A CBC by Sysmex XT-2000iV revealed a moderate normocytic normochromic anemia. Red blood cells counted by optic flow cytometry (RBC-O) resulted in a higher value than using electrical impedance (RBC-I). The relative reticulocyte count based on RNA content and size was 35.3%, while the manual reticulocyte count was < 1%. The WBC count of 1,562,680 cells/μL was accompanied by a flag. Manual counts for RBC and WBC using the Bürker chamber confirmed the Sysmex impedance results. Finally the manual PCV was lower than HCT by Sysmex. While Sysmex XT can differentiate between RBC and WBC by impedance, even in the face of extreme lymphocytosis due to CLL, RBC-O can be affected by bias, resulting in falsely increased RBC and reticulocyte numbers. Overestimation of RBC-O may be due to incorrect Sysmex classification of leukemic cells or their fragments as reticulocytes. This phenomenon is known as pseudoreticulocytosis and can lead to misinterpretation of regenerative anemia. On the other side PCV can be affected by bias in CLL due to the trapping of RBC in the buffy coat, resulting in a pink hue in the separation area. As HGB concentration is not affected by flow cytometric or other cell-related artifacts it may represent the most reliable variable to assess the degree of anemia in cases of CLL.

  18. Interleukin-2 dose, blood monocyte and CD25+ lymphocyte counts as predictors of clinical response to interleukin-2 therapy in patients with renal cell carcinoma

    DEFF Research Database (Denmark)

    Hermann, G G; Geertsen, P F; von der Maase, H

    1991-01-01

    The purpose of this study was to determine immunological parameters in the peripheral blood that correlate with the clinical effect of interleukin-2 (IL-2) in patients with metastatic renal cell cancer. A group of 26 patients with metastatic renal cell cancer underwent IL-2 treatment using a 36-day...... schedule with continuous intravenous IL-2 infusion (3 x 10(6) units m-2 day-1) administered from days 1 to 5 and days 12 to 16. The white blood cell count and the absolute and relative number of neutrophils, lymphocytes, eosinophils and monocytes were recorded six times in peripheral blood during...

  19. Early lymphocyte recovery predicts superior overall survival after unmanipulated haploidentical blood and marrow transplant for myelodysplastic syndrome and acute myeloid leukemia evolving from myelodysplastic syndrome.

    Science.gov (United States)

    Chang, Ying-Jun; Zhao, Xiang-Yu; Xu, Lan-Ping; Liu, Dai-Hong; Liu, Kai-Yan; Chen, Yu-Hong; Wang, Yu; Zhang, Xiao-Hui; Zhao, Xiao-Su; Han, Wei; Chen, Huan; Wang, Feng-Rong; Lv, Meng; Huang, Xiao-Jun

    2013-12-01

    We investigated whether early lymphocyte recovery, after unmanipulated, haploidentical, blood and marrow transplant (HBMT), affected clinical outcomes in 78 patients with myelodysplastic syndrome (MDS) or acute myeloid leukemia evolving from MDS. Lymphocyte recovery was based on the absolute lymphocyte count on day 30 (ALC-30). Patients with high ALC-30 (≥ 300 cells/μL) had lower relapse rates (13.8% vs. 35.5%, p = 0.049) and lower incidence of bacterial infections (3.4% vs. 25.8%, p = 0.015) than those with low ALC-30 values. Multivariate analysis showed that a high ALC-30 was associated with improved overall survival (OS, hazard ratio [HR]: 0.099, 95% confidence interval [CI]: 0.029-0.337; p leukemia-free survival (HR: 0.245, 95% CI: 0.112-0.539; p after unmanipulated HBMT.

  20. Ligation of MHC class I molecules on peripheral blood T lymphocytes induces new phenotypes and functions

    DEFF Research Database (Denmark)

    Bregenholt, S; Röpke, M; Skov, S;

    1996-01-01

    Microgram concentrations of immobilized anti-MHC class I (MHC-I) Ab induced proliferation of resting CD3+ T cells from peripheral blood. In contrast, soluble Ab did not activate T cells. Exposure of T cells to immobilized anti-MHC-I Ab for only 24 h was followed by proliferation and development....... These data clearly demonstrate that ligation of the MHC-I complex on T cells may induce both positive and negative signals. Since the physiologic ligands for MHC-I molecules are TCR and the CD8 molecules, our data may suggest that MHC-I molecules are instrumental in cellular interactions between T cells....

  1. Immunophenotyping of peripheral blood lymphocytes in children and adolescents with Hashimoto`s 1 thyroiditis from the areas contaminated as aa result of the Chernobyl accident

    Energy Technology Data Exchange (ETDEWEB)

    Molostvov, G.S. [Research Inst. of Radiation Medicine, Minsk (Belarus)

    1996-12-31

    m6o-color immunophenotyping of lymphocyte subsets using a lysed whole blood method was performed in 46 children and adolescents with Hashimoto`s thyroiditis (HT) from contaminated areas and in 18 children with HT from `pure` areas of Belarus. 46 healthy children of matched age and sex distribution were used as control group. Analysis of lymphocyte subsets in children with HT living in contaminated areas showed a considerable decrease in the levels of total lymphocytes, CD8+T cells (T-suppressors), total B cells, and CD5+B cells together with an activation of NK and CD56+, CD8+CD57+T cells (T-killers not restricted by HLA antigens). The study of cellular immunity in children with different doses of incorporated radionuclids revealed that prolonged influence of very small doses of ionizing radiation resulted in significant changes in lymphocyte subsets; interestingly, B cell subsets were the most sensitive to such influence while radiosensitivity of T-killers and NK was the lowest. These changes were the greatest in children with HT with the highest dose of incorporated radionuclides. Significant correlation between the levels of main lymphocyte subsets and the doses of accumulated radionuclids observed in this study also indicated that their relation was dose-dependent.(orig.)

  2. Antibody to histo-blood group A antigen neutralizes HIV produced by lymphocytes from blood group A donors but not from blood group B or O donors

    DEFF Research Database (Denmark)

    Arendrup, M; Hansen, J E; Clausen, H;

    1991-01-01

    not inhibit the HTLV-IIIB/lyB or the HTLV-IIIB/lyO isolate. Specificity of the MAb-mediated inhibition was shown using A-antigen (tetrasaccharide). Thus, HIV infection of PBMC from donors with blood type A appears to induce expression of host-cell-encoded carbohydrate blood group A epitope on HIV which can...

  3. Radio-protective effect of cinnamic acid, a phenolic phytochemical, on genomic instability induced by X-rays in human blood lymphocytes in vitro.

    Science.gov (United States)

    Cinkilic, Nilufer; Tüzün, Ece; Çetintaş, Sibel Kahraman; Vatan, Özgür; Yılmaz, Dilek; Çavaş, Tolga; Tunç, Sema; Özkan, Lütfi; Bilaloğlu, Rahmi

    2014-08-01

    The present study was designed to determine the protective activity of cinnamic acid against induction by X-rays of genomic instability in normal human blood lymphocytes. This radio-protective activity was assessed by use of the cytokinesis-block micronucleus test and the alkaline comet assay, with human blood lymphocytes isolated from two healthy donors. A Siemens Mevatron MD2 (Siemens AG, USA, 1994) linear accelerator was used for the irradiation with 1 or 2 Gy. Treatment of the lymphocytes with cinnamic acid prior to irradiation reduced the number of micronuclei when compared with that in control samples. Treatment with cinnamic acid without irradiation did not increase the number of micronuclei and did not show a cytostatic effect in the lymphocytes. The results of the alkaline comet assay revealed that cinnamic acid reduces the DNA damage induced by X-rays, showing a significant radio-protective effect. Cinnamic acid decreased the frequency of irradiation-induced micronuclei by 16-55% and reduced DNA breakage by 17-50%, as determined by the alkaline comet assay. Cinnamic acid may thus act as a radio-protective compound, and future studies may focus on elucidating the mechanism by which cinnamic acid offers radioprotection.

  4. Incidence of micronuclei in human peripheral blood lymphocytes exposed to modulated and unmodulated 2450 MHz radiofrequency fields.

    Science.gov (United States)

    Vijayalaxmi; Reddy, Abhishek B; McKenzie, Raymond J; McIntosh, Robert L; Prihoda, Thomas J; Wood, Andrew W

    2013-10-01

    Peripheral blood samples from four healthy volunteers were collected and aliquots were exposed in vitro for 2 h to either (i) modulated (wideband code division multiple access, WCDMA) or unmodulated continuous wave (CW) 2450 MHz radiofrequency (RF) fields at an average specific absorption rate of 10.9 W/kg or (ii) sham-exposed. Aliquots of the same samples that were exposed in vitro to an acute dose of 1.5 Gy ionizing gamma-radiation (GR) were used as positive controls. Half of the aliquots were treated with melatonin (Mel) to investigate if such treatment offers protection to the cells from the genetic damage, if any, induced by RF and GR. The cells in all samples were cultured for 72 h and the lymphocytes were examined to determine the extent of genetic damage assessed from the incidence of micronuclei (MN). The results indicated the following: (i) the incidence of MN was similar in incubator controls, and those exposed to RF/sham and Mel alone; (ii) there were no significant differences between WCDMA and CW RF exposures; (iii) positive control cells exposed to GR alone exhibited significantly increased MN; and (iv) Mel treatment had no effect on cells exposed to RF and sham, while such treatment significantly reduced the frequency of MN in GR-exposed cells.

  5. Effects of Sterigmatocystin, Deoxynivalenol and Aflatoxin G1 on Apoptosis of Human Peripheral Blood Lymphocytes in vitro

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Objective To explore the effects of Sterigmatocystin (ST), Deoxynivalenol (DON) and Aflatoxin G1 (AFG1) on apoptosis of human peripheral blood lymphocytes (HPBLs) in vitro and thus to further elucidate the putative roles of these three mycotoxins on human immunosystem. Methods The effects of ST, DON and AFG1 on apoptosis of HPBLs were studied with cell culture, flow cytometric (FCM) DNA analysis and DNA agarose gel electrophoresis. Results DNA agarose gel electrophoresis results showed the characteristic "ladder" pattern of apoptosis in HPBLs treated with ST, DON and AFG1. Flow cytometric DNA analysis revealed that typical subdiploid peaks of apoptosis in DNA histogram could be seen in all groups treated with the three mycotoxins. Significant time-effect and dose-effect relationships were found between the apoptosis rates and treatment time as well as concentrations of the three mycotoxins. Conclusion ST, DON and AFG1 can induce apoptosis of HPBLs in vitro and may have some negative effects on human immunosystem.

  6. [Proliferative activity parameters and their correlation with genetic damage of blood lymphocytes during cultivation under the conditions of cytokinetic block].

    Science.gov (United States)

    Ingel', F I; Iurchenko, V V; Gus'kov, A S; Krivtsova, E K; Iurtseva, N A

    2006-01-01

    The subjects of the study were 15 volunteers aged 22 to 25 years, who underwent 25 air ionization sessions. The effects of genome instability were evaluated, and correlations between indicators of genome damage (lesions of micronuclei and nucleoplasmatic bridges) and parameters of proliferative and replicative activity (mitotic index, proliferative pool, the fraction of rapidly dividing cells, and replication index) of blood lymphocytes in the culture were studied. In order to establish the associations between the parameters, the parallel cultures were exposed to 0.07 mM of the standard mutagen MNNG during 5 hours. The study showed that the course of air ionization did not induce the micronuclei and nucleoplasmatic bridges in binuclear cells, but increased proliferative cell activity. This effect was accompanied by an increase in the fraction of rapidly dividing cells among all the dividing cells, and an increase in the dispersion of all proliferation parameters. MNNG induced a constant level of micronuclei in binuclear cells during the whole course, but not before the beginning of air ionization. The changes in the parameter "the fraction of dividing cells" (proliferative pool) were the most prominent manifestation of the suppression of proliferation by MNNG. MNNG loading inhibited the formation of binuclear cells most of all. The results demonstrate a non-random character of the correlation between the level of micronuclei in binuclear cells and proliferative activity parameters during cell cultivation under the conditions of cytokinetic block.

  7. The dopaminergic system in peripheral blood lymphocytes: from physiology to pharmacology and potential applications to neuropsychiatric disorders.

    Science.gov (United States)

    Buttarelli, Francesca R; Fanciulli, Alessandra; Pellicano, Clelia; Pontieri, Francesco E

    2011-06-01

    Besides its action on the nervous system, dopamine (DA) plays a role on neural-immune interactions. Here we review the current evidence on the dopaminergic system in human peripheral blood lymphocytes (PBL). PBL synthesize DA through the tyrosine-hydroxylase/DOPA-decarboxylase pathway, and express DA receptors and DA transporter (DAT) on their plasma membrane. Stimulation of DA receptors on PBL membrane contributes to modulate the development and initiation of immune responses under physiological conditions and in immune system pathologies such as autoimmunity or immunodeficiency.The characterization of DA system in PBL gave rise to a further line of research investigating the feasibility of PBL as a cellular model for studying DA derangement in neuropsychiatric disorders. Several reports showed changes of the expression of DAT and/or DA receptors in PBL from patients suffering from several neuropsychiatric disorders, in particular parkinsonian syndromes, schizophrenia and drug- or alcohol-abuse. Despite some methodological and theoretical limitations, these findings suggest that PBL may prove a cellular tool with which to identify the derangement of DA transmission in neuropsychiatric diseases, as well as to monitor the effects of pharmacological treatments.

  8. Construction of a human recombinant polyclonal Fab fragment antibody library using peripheral blood lymphocytes of snake bitten victims

    Directory of Open Access Journals (Sweden)

    Motedayen, M.H.

    2015-12-01

    Full Text Available Human snake bitten poisoning is a serious threat in many tropical and subtropical countries such as Iran. The best acceptable treatment of envenomated humans is antivenoms; however they have a series of economic and medical problems and need more improvements. In this study a combinatorial human immunoglobulin gene library against some of Iranian snakes venoms was constructed. Total RNA prepared from peripheral blood lymphocytes of two recovered snake victims. RT-PCR was used for cDNA synthesis and amplification of the heavy (Fd segment and kappa light chains of IgG antibody. After digestion of the heavy chain with SpeI and XhoI and light chain with XbaI and SacI enzymes, inserted successively into the cloning vector pComb3x, and then recombinant vector transformed to TG1 bacteria to construct the Fab library. For determination insertion rate of Fab segment into cloning vector, plasmids of 12 clones of library were extracted and digested with SfiI. Length of amplified Fd and κ chains, were 650 - 750 bp. Primary library size was determined to contain 4.9×105 members out of which half of them contained the same size of Fab fragment. This result is comparable to some researchers and shows that this method could be appropriate tool for the production of human polyclonal Fab fragment antibodies for management of poisonous snake bitted victims.

  9. Cytogenetic genotoxic investigation in peripheral blood lymphocytes of subjects with dental composite restorative filling materials.

    Science.gov (United States)

    Pettini, F; Savino, M; Corsalini, M; Cantore, S; Ballini, A

    2015-01-01

    Dental composite resins are biomaterials commonly used to aesthetically restore the structure and function of teeth impaired by caries, erosion, or fracture. Residual monomers released from resin restorations as a result of incomplete polymerization processes interact with living oral tissues. The objective of this study was to evaluate the genotoxicity of a common dental composite material (Enamel Plus-HFO), in subjects with average 13 filled teeth with the same material, compared to a control group (subjects having neither amalgam nor composite resin fillings). Genotoxicity assessment of composite materials was carried out in vitro in human peripheral blood leukocytes using sister-chromatid exchange (SCE) and chromosomal aberrations (CA) cytogenetic tests. The results of correlation and multiple regression analyses confirmed the absence of a relationship between SCE/cell, high frequency of SCE(HFC) or CA frequencies and exposure to dental composite materials. These results indicate that composite resins used for dental restorations differ extensively in vivo in their cytotoxic and genotoxic potential and in their ability to affect chromosomal integrity, cell-cycle progression, DNA replication and repair.

  10. Rapid gene expression changes in peripheral blood lymphocytes upon practice of a comprehensive yoga program.

    Directory of Open Access Journals (Sweden)

    Su Qu

    Full Text Available One of the most common integrative medicine (IM modalities is yoga and related practices. Previous work has shown that yoga may improve wellness in healthy people and have benefits for patients. However, the mechanisms of how yoga may positively affect the mind-body system are largely unknown. Here we have assessed possible rapid changes in global gene expression profiles in the peripheral blood mononuclear cells (PBMCs in healthy people that practiced either a comprehensive yoga program or a control regimen. The experimental sessions included gentle yoga postures, breathing exercises, and meditation (Sudarshan Kriya and Related Practices--SK&P compared with a control regimen of a nature walk and listening to relaxing music. We show that the SK&P program has a rapid and significantly greater effect on gene expression in PBMCs compared with the control regimen. These data suggest that yoga and related practices result in rapid gene expression alterations which may be the basis for their longer term cell biological and higher level health effects.

  11. Rapid gene expression changes in peripheral blood lymphocytes upon practice of a comprehensive yoga program.

    Science.gov (United States)

    Qu, Su; Olafsrud, Solveig Mjelstad; Meza-Zepeda, Leonardo A; Saatcioglu, Fahri

    2013-01-01

    One of the most common integrative medicine (IM) modalities is yoga and related practices. Previous work has shown that yoga may improve wellness in healthy people and have benefits for patients. However, the mechanisms of how yoga may positively affect the mind-body system are largely unknown. Here we have assessed possible rapid changes in global gene expression profiles in the peripheral blood mononuclear cells (PBMCs) in healthy people that practiced either a comprehensive yoga program or a control regimen. The experimental sessions included gentle yoga postures, breathing exercises, and meditation (Sudarshan Kriya and Related Practices--SK&P) compared with a control regimen of a nature walk and listening to relaxing music. We show that the SK&P program has a rapid and significantly greater effect on gene expression in PBMCs compared with the control regimen. These data suggest that yoga and related practices result in rapid gene expression alterations which may be the basis for their longer term cell biological and higher level health effects.

  12. Cytogenetic damage in human blood lymphocytes exposed in vitro to radon

    Energy Technology Data Exchange (ETDEWEB)

    Hamza, V. Zareena [Radiological Safety Division, Indira Gandhi Centre for Atomic Research (IGCAR), Kalpakkam-603 102, Tamilnadu (India); Mohankumar, Mary N. [Radiological Safety Division, Indira Gandhi Centre for Atomic Research (IGCAR), Kalpakkam-603 102, Tamilnadu (India)], E-mail: marynmk@igcar.gov.in

    2009-02-10

    The effect of radon in inducing DNA damage was investigated in vitro by two well-established cytogenetic assays. Blood samples were irradiated with radon using a novel irradiation assembly. Doses varied between 0 and 127 mGy for chromosome aberration (CA) assay and 0 and 120 mGy for cytokinesis blocked micronucleus (CBMN) assay. Dose-rates varied between 0.000054 and 0.708 mGy/min. After the irradiation period of 3 h, excess radon gas was released and cultures were initiated using standard procedures. Chromosome aberrations such as dicentrics, excess acentric fragments, acentric rings, centric rings, chromatid breaks were observed. Micronuclei, nucleoplasmic bridges and nuclear buds were scored by the CBMN assay. A significant increase in the frequency of dicentrics, excess acentric fragments and centric rings was observed with increasing radon dose, whereas total acentric rings plus double minute and chromatid breaks/cell were not significantly elevated. In CBMN assay, the frequency of micronuclei was found to be significantly raised whereas that of nucleoplasmic bridges and nuclear buds were not. Nucleoplasmic bridges and nuclear buds tended to increase with dose but did not achieve statistical significance. There was a strong positive correlation between nucleoplasmic bridges and dicentrics (P < 0.028) or rings (P < 0.0001) and between micronuclei and acentric fragments (P < 0.0005). The study shows that radon is capable of inducing significant chromosome damage at very low doses and dose-rates.

  13. Combined Treatment with Low Concentrations of Decitabine and SAHA Causes Cell Death in Leukemic Cell Lines but Not in Normal Peripheral Blood Lymphocytes

    Directory of Open Access Journals (Sweden)

    Barbora Brodská

    2013-01-01

    Full Text Available Epigenetic therapy reverting aberrant acetylation or methylation offers the possibility to target preferentially tumor cells and to preserve normal cells. Combination epigenetic therapy may further improve the effect of individual drugs. We investigated combined action of demethylating agent decitabine and histone deacetylase inhibitor SAHA (Vorinostat on different leukemic cell lines in comparison with peripheral blood lymphocytes. Large decrease of viability, as well as huge p21WAF1 induction, reactive oxygen species formation, and apoptotic features due to combined decitabine and SAHA action were detected in leukemic cell lines irrespective of their p53 status, while essentially no effect was observed in response to the combined drug action in normal peripheral blood lymphocytes of healthy donors. p53-dependent apoptotic pathway was demonstrated to participate in the wtp53 CML-T1 leukemic cell line response, while significant influence of reactive oxygen species on viability decrease has been detected in p53-null HL-60 cell line.

  14. Interleukin-2 dose, blood monocyte and CD25+ lymphocyte counts as predictors of clinical response to interleukin-2 therapy in patients with renal cell carcinoma

    DEFF Research Database (Denmark)

    Hermann, G G; Geertsen, P F; von der Maase, H;

    1991-01-01

    schedule with continuous intravenous IL-2 infusion (3 x 10(6) units m-2 day-1) administered from days 1 to 5 and days 12 to 16. The white blood cell count and the absolute and relative number of neutrophils, lymphocytes, eosinophils and monocytes were recorded six times in peripheral blood during...... the treatment. Also the blood counts of T cell and NK cell subsets and cells expressing the T cell activation markers IL-2R alpha and VLA-1 were measured. The lymphokine-activated killer (LAK) cell cytotoxicity was measured either with or without additional in vitro stimulation by IL-2. Multivariate statistical...... analysis showed that the clinical responses were related to the administered dose of IL-2, to a low number of blood cells expressing IL-2 receptors and to a reduction in the blood monocyte count (P less than 0.05)....

  15. Day 100 Peripheral Blood Absolute Lymphocyte/Monocyte Ratio and Survival in Classical Hodgkin's Lymphoma Postautologous Peripheral Blood Hematopoietic Stem Cell Transplantation

    Directory of Open Access Journals (Sweden)

    Luis F. Porrata

    2013-01-01

    Full Text Available Day 100 prognostic factors of postautologous peripheral blood hematopoietic stem cell transplantation (APBHSCT to predict clinical outcome in classical Hodgkin lymphoma (cHL patients have not been evaluated. Thus, we studied if the day 100 peripheral blood absolute lymphocyte/monocyte ratio (Day 100 ALC/AMC affects clinical outcomes by landmark analysis from day 100 post-APBHSCT. Only cHL patients achieving a complete remission at day 100 post-APBHSCT were studied. From 2000 to 2010, 131 cHL consecutive patients qualified for the study. The median followup from day 100 was 4.1 years (range: 0.2–12.3 years. Patients with a Day 100 ALC/AMC ≥ 1.3 experienced superior overall survival (OS and progression-free survival (PFS compared with Day 100 ALC/AMC < 1.3 (from day 100: OS, median not reached versus 2.8 years; 5 years OS rates of 93% (95% CI, 83%–97% versus 35% (95% CI, 19%–51%, resp., P<0.0001; from day 100: PFS, median not reached versus 1.2 years; 5 years PFS rates of 79% (95% CI, 69%–86% versus 27% (95% CI, 14%–45%, resp., P<0.0001. Day ALC/AMC ratio was an independent predictor for OS and PFS. Thus, Day 100 ALC/AMC ratio is a simple biomarker that can help to assess clinical outcomes from day 100 post-APBHSCT in cHL patients.

  16. Thymic hormonal activity on human peripheral blood lymphocytes, in vitro. V. Effect on induction of lymphocytotoxicity.

    Science.gov (United States)

    Shoham, J; Cohen, M

    1983-01-01

    Thymic hormonal effect on lymphocytotoxicity induced in vitro and its target specificity were tested using peripheral blood mononuclear cells (PBMC) of healthy subjects. PBMC were treated by the thymic extract TP-1, a similarly prepared spleen extract (SE) or medium only (1 h, 37 degrees C) and then induced to express cytotoxic activity by exposure to allogeneic tumor cells in mixed cultures or by Con A stimulation. The cytotoxicity developed after several days in culture was assayed on 51Cr labelled tumor cells. TP-1 caused a significant mean enhancement of cytotoxicity induced and assayed on Raji lymphoma cells (mean % specific lysis, 31.5 +/- 2.9 without TP-1 and 53.7 +/- 3.6 with TP-1; n = 42; p less than 0.01). The scatter of individual responses to TP-1 was wide, however, and included also some cases of TP-1 induced suppression. Similar wide scatter of TP-1 effects with emphasis on TP-1 induced enhancement was observed with other tumor cell lines or with Con A as inducers. Usually, SE had no effect on induced cytotoxicity. Target selectivity (specificity) of induced cytotoxicity was tested by induction and assay on several tumor cell lines with crossing over, as well as by cold competition assay. When target selectivity was present, it was not masked by TP-1 induced enhancement. Moreover, in some cases, target selectivity became more pronounced after TP-1 treatment. However, TP-1 enhanced also Con A induced non-specific cytotoxicity. No effect of TP-1 on natural killer cell activity of fresh PBMC could be demonstrated. It is suggested that both selective cytotoxicity (T-cell dependent) and non-selective one maybe modulated directly by TP-1 and indirectly by TP-1 modified secondary interactions in culture. This profound regulatory effects could be demonstrated in the PBMC of immune-intact healthy adults.

  17. Hepatitis C virus (HCV)-specific in vitro antibody secretion by peripheral blood lymphocytes: correlation with progression of disease and HCV RNA in HCV antibody-positive patients.

    OpenAIRE

    Ducos, J.; Bianchi-Mondain, A M; Francois, M.; Boisset, M; Vendrell, J P; Barin, F; Serre, A; Larrey, D

    1994-01-01

    Hepatitis C virus-specific in vitro antibody production (HCV IVAP) by peripheral blood lymphocytes in 53 HCV antibody-positive patients was investigated in comparison with alanine aminotransferase (ALT) levels and HCV RNA in serum samples. All 29 HCV IVAP-positive patients were HCV RNA positive; 26 had elevated ALT levels. Among the 24 HCV IVAP-negative patients, 16 were HCV RNA negative, with 12 presenting normal ALT values. These data indicate that HCV IVAP results are highly correlated (P ...

  18. Over-Expression of Dopamine D2 Receptor and Inwardly Rectifying Potassium Channel Genes in Drug-Naive Schizophrenic Peripheral Blood Lymphocytes as Potential Diagnostic Markers

    OpenAIRE

    Ágnes Zvara; György Szekeres; Zoltán Janka; Kelemen, János Z.; Csongor Cimmer; Miklós Sántha; Puskás, László G.

    2005-01-01

    Schizophrenia is one of the most common neuropsychiatric disorders affecting nearly 1% of the human population. Current diagnosis of schizophrenia is based on complex clinical symptoms. The use of easily detectable peripheral molecular markers could substantially help the diagnosis of psychiatric disorders. Recent studies showed that peripheral blood lymphocytes (PBL) express subtypes of D1 and D2 subclasses of dopamine receptors. Recently, dopamine receptor D3 (DRD3) was found to be over-exp...

  19. [Cytogenetic analysis of cosmonaut's blood lymphocytes exposed to low space radiation doses and its place in clinical-physiological health monitoring].

    Science.gov (United States)

    Fedorenko, B S; Snigireva, G P; Ivanov, A A

    2010-01-01

    The article discusses the informative significance and pertinence of cytogenetic analysis of peripheral blood lymphocytes to the clinical-physiological investigations of cosmonauts conducted before and after space missions (SM) of varying duration. The authors point out high sensitivity of the technique, relative simplicity and affordability of the analysis of instable chromosomal aberrations including such radiation effect markers as dicentrics and centric rings. Cytogenetic investigation of cosmonauts could be used to estimate ionizing radiation dose, and to predict delayed effects.

  20. Study on the blood-borne virus co-infection and T lymphocyte subset among intravenous drug users

    Institute of Scientific and Technical Information of China (English)

    Jian-Rong Li; Rui-Yu Gong; Kun-Lun Tian; Jing Wang; Yi-Xin Wang; Han-Ju Huang

    2007-01-01

    AIM: To investigate the features of various bloodborne virus infections and co-infection in intravenous drug users (IDUs), and to examine the correlation of T lymphocyte subsets with virus co-infection.METHODS: Four hundred and six IDUs without any clinical manifestation of hepatitis and 102 healthy persons were enrolled in this study. HBV-DNA and HCV-RNA were detected by fluorescence quantitative PCR. HBsAg, HBeAg, anti-HBc, anti-HCV, HDV-Ag, anti-HGV, anti-HIV, and HCMV-IgM were assayed by enzyme-linked immunosorbent assay (ELISA) and immunochromatographic tests. The levels of Th1 and Th2 cytokines were measured by ELISA and radioactive immune assay (RIA). The T lymphocyte subpopulation was detected by using fluorescence immunoassay. The similar indices taken from the healthy persons served as controls.RESULTS: The viral infection rate among IDUs was 36.45% for HBV, 69.7% for HCV, 47.3% for HIV, 2.22% for HDV, 1.97% for HGV, and 3.45% for HCMV. The co-infection rate of blood-borne virus was detected in 255 of 406 (62.81%) IDUs. More than 80% (161/192) of subjects infected with HIV were co-infected with the other viruses, such as HBV, HCV. In contrast, among the controls, the infection rate was 17.65% for HBV and 0% for the other viruses. Our investigation showed that there was a profound decrease in the proportion of CD4/CD8 and the percentage of CD3 and CD4, but not in the percentage of CD8. The levels of PHA-induced cytokines (IFN-γ and IL-4) and serum IL-2 were obviously decreased in IDUs. On the other hand, the level of serum IL-4 was increased. The level of IFN-γ and the percentage of CD4 were continuously decreased when the IDUs were infected with HIV or HIV co-infection. IDUs with HIV and HBV co-infection was 15.1% (29/192). Of those 29 IDU with HIV and HBV co-infection, 51.72% (15/29) and 37.93% (11/29) were HBV-DNA-positive and HBeAg-positive, respectively. But, among IDUs without HIV infection, only 1.68% (2/119) of cases were HBV

  1. Phenotypic and Functional Characterization of Peripheral Blood Lymphocytes from Various Age- and Sex-Specific Groups of Owl Monkeys (Aotus nancymaae).

    Science.gov (United States)

    Nehete, Pramod N; Nehete, Bharti P; Chitta, Sriram; Williams, Lawrence E; Abee, Christian R

    2017-02-01

    Owl monkeys (Aotus nancymaae) are New World NHP that serve an important role in vaccine development and as a model for human disease conditions such as malaria. Despite the past contributions of this animal model, limited information is available about the phenotype and functional properties of peripheral blood lymphocytes in reference to sex and age. Using a panel of human antibodies and a set of standardized human immune assays, we identified and characterized various peripheral blood lymphocyte subsets, evaluated the immune functions of T cells, and analyzed cytokines relative to sex and age in healthy owl monkeys. We noted age- and sex-dependent changes in CD28+ (an essential T cell costimulatory molecule) and CD95+ (an apoptotic surface marker) T cells and various levels of cytokines in the plasma. In immune assays of freshly isolated peripheral blood mononuclear cells, IFNγ and perforin responses were significantly higher in female than in male monkeys and in young adults than in juvenile and geriatric groups, despite similar lymphocyte (particularly T cell) populations in these groups. Our current findings may be useful in exploring Aotus monkeys as a model system for the study of aging, susceptibility to infectious diseases, and age-associated differences in vaccine efficacy, and other challenges particular to pediatric and geriatric patients.

  2. Antibody to histo-blood group A antigen neutralizes HIV produced by lymphocytes from blood group A donors but not from blood group B or O donors

    DEFF Research Database (Denmark)

    Arendrup, M; Hansen, J E; Clausen, H;

    1991-01-01

    for virus neutralization by the monoclonal antibody (MAb) AH16 directed against the blood group A epitope. MAb AH16 was previously shown to inhibit cell-free virus infection using HTLV-IIIB propagated in H9 cells. AH16 showed a concentration-dependent inhibition of the HTLV-IIIB/lyA isolate but did...... not inhibit the HTLV-IIIB/lyB or the HTLV-IIIB/lyO isolate. Specificity of the MAb-mediated inhibition was shown using A-antigen (tetrasaccharide). Thus, HIV infection of PBMC from donors with blood type A appears to induce expression of host-cell-encoded carbohydrate blood group A epitope on HIV which can......Three virus isolates HTLV-IIIB/lyA, HTLV-IIIB/lyB and HTLV-IIIB/lyO, obtained by passaging and propagating the HTLV-IIIB/H9 isolate in three separate cultures of mixed peripheral blood mononuclear cells (PBMC) from donors of blood type A, B or O, respectively, were tested for susceptibility...

  3. Evaluation of Genotoxic and Cytotoxic Effects in Human Peripheral Blood Lymphocytes Exposed In Vitro to Neonicotinoid Insecticides News

    Directory of Open Access Journals (Sweden)

    María Elena Calderón-Segura

    2012-01-01

    Full Text Available Calypso (thiacloprid, Poncho (clothianidin, Gaucho (imidacloprid, and Jade (imidacloprid are commercial neonicotinoid insecticides, a new class of agrochemicals in México. However, genotoxic and cytotoxic studies have not been performed. In the present study, human peripheral blood lymphocytes (PBL were exposed in vitro to different concentrations of the four insecticides. The genotoxic and cytotoxic effects were evaluated using the alkaline comet and trypan blue dye exclusion assays. DNA damage was evaluated using two genotoxicity parameters: tail length and comet frequency. Exposure to 9.5×10-6 to 5.7×10-5 M Jade; 2.8×10-4 to 1.7×10-3 M Gaucho; 0.6×10-1 to 1.4×10-1 M Calypso; 1.2×10-1 to 9.5×10-1 M Poncho for 2 h induced a significant increase DNA damage with a concentration-dependent relationship. Jade was the most genotoxic of the four insecticides studied. Cytotoxicity was observed in cells exposed to 18×10-3 M Jade, 2.0×10-3 M Gaucho, 2.0×10-1 M Calypso, 1.07 M Poncho, and cell death occurred at 30×10-3 M Jade, 3.3×10-3 M Gaucho, 2.8×10-1 M Calypso, and 1.42 M Poncho. This study provides the first report of genotoxic and cytotoxic effects in PBL following in vitro exposure to commercial neonicotinoid insecticides.

  4. Evaluation of Genotoxic and Cytotoxic Effects in Human Peripheral Blood Lymphocytes Exposed In Vitro to Neonicotinoid Insecticides News

    Science.gov (United States)

    Calderón-Segura, María Elena; Gómez-Arroyo, Sandra; Villalobos-Pietrini, Rafael; Martínez-Valenzuela, Carmen; Carbajal-López, Yolanda; Calderón-Ezquerro, María del Carmen; Cortés-Eslava, Josefina; García-Martínez, Rocío; Flores-Ramírez, Diana; Rodríguez-Romero, María Isabel; Méndez-Pérez, Patricia; Bañuelos-Ruíz, Enrique

    2012-01-01

    Calypso (thiacloprid), Poncho (clothianidin), Gaucho (imidacloprid), and Jade (imidacloprid) are commercial neonicotinoid insecticides, a new class of agrochemicals in México. However, genotoxic and cytotoxic studies have not been performed. In the present study, human peripheral blood lymphocytes (PBL) were exposed in vitro to different concentrations of the four insecticides. The genotoxic and cytotoxic effects were evaluated using the alkaline comet and trypan blue dye exclusion assays. DNA damage was evaluated using two genotoxicity parameters: tail length and comet frequency. Exposure to 9.5 × 10−6 to 5.7 × 10−5 M Jade; 2.8 × 10−4 to 1.7 × 10−3 M Gaucho; 0.6 × 10−1 to 1.4 × 10−1 M Calypso; 1.2 × 10−1 to 9.5 × 10−1 M Poncho for 2 h induced a significant increase DNA damage with a concentration-dependent relationship. Jade was the most genotoxic of the four insecticides studied. Cytotoxicity was observed in cells exposed to 18 × 10−3 M Jade, 2.0 × 10−3 M Gaucho, 2.0 × 10−1 M Calypso, 1.07 M Poncho, and cell death occurred at 30 × 10−3 M Jade, 3.3 × 10−3 M Gaucho, 2.8 × 10−1 M Calypso, and 1.42 M Poncho. This study provides the first report of genotoxic and cytotoxic effects in PBL following in vitro exposure to commercial neonicotinoid insecticides. PMID:22545045

  5. Early effects of low dose 12C6+ ion or X-ray irradiation on human peripheral blood lymphocytes

    Science.gov (United States)

    Chen, Yingtai; Li, Yumin; Zhang, Hong; Xie, Yi; Chen, Xuezhong; Ren, Jinyu; Zhang, Xiaowei; Zhu, Zijiang; Liu, Hongliang; Zhang, Yawei

    2010-04-01

    The aim of this study was to estimate the acute effects of low dose 12C6+ ions or X-ray radiation on human immune function. The human peripheral blood lymphocytes (HPBL) of seven healthy donors were exposed to 0.05 Gy 12C6+ ions or X-ray radiation and cell responses were measured at 24 h after exposure. The cytotoxic activities of HPBL were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT); the percentages of T and NK cells subsets were detected by flow cytometry; mRNA expression of interleukin (IL)-2, tumor necrosis factor (TNF)-α and interferon (IFN)-γ were examined by real time quantitative RT-PCR (qRT-PCR); and these cytokines protein levels in supernatant of cultured cells were assayed by enzyme-linked immunosorbent assays (ELISA). The results showed that the cytotoxic activity of HPBL, mRNA expression of IL-2, IFN-γ and TNF-α in HPBL and their protein levels in supernatant were significantly increased at 24 h after exposure to 0.05 Gy 12C6+ ions radiation and the effects were stronger than observed for X-ray exposure. However, there was no significant change in the percentage of T and NK cells subsets of HPBL. These results suggested that 0.05 Gy high linear energy transfer (LET) 12C6+ radiation was a more effective approach to host immune enhancement than that of low LET X-ray. We conclude that cytokines production might be used as sensitive indicators of acute response to LDI.

  6. Comparison of chromosome aberrations in peripheral blood lymphocytes from people occupationally exposed to ionizing and radiofrequency radiation.

    Directory of Open Access Journals (Sweden)

    Lalić H

    2001-04-01

    Full Text Available The genotoxic effects of occupational exposure to ionizing and non-ionizing radiation were investigated in 25 physicians and nurses working in hospitals and in 20 individuals working at radio-relay stations. Examination was conducted by chromosome aberration analysis of peripheral blood lymphocytes. The data showed that total number of chromosome aberrations in people exposed to ionizing and radio-frequency radiation (4.08 +/- 0.37 and 4.35 +/- 0.5 on 200 scored metaphases, respectively were almost equally higher than those of non-irradiated subjects. The increase was in proportion to the number of individuals having more that 5-aberration/200 metaphases. Acentric fragments comprised the most frequently seen type of aberration. The average numbers in examined groups (11.8 x 10(-3 and 14.8 x 10(-3 per cell, respectively, were significantly higher than 4.2 x 10(-3, which was observed in controls, unexposed individuals. Dicentric fragments were also frequent (4.8 x 10(-3 and 6.25 x 10(-3, respectively, vs. 0.52 x 10(-3 in control. In contrast, the frequency of chromatid breaks increased only after ionizing radiation (3.8 x 10(-3 vs. 0.26 x 10(-3 in control. A positive correlation between the total number of chromosome aberrations and cumulative 6-years dosage was also found. The data emphasized the dangerous effects of prolonged exposure to both types of radiation and indicated that chromosomal aberration analysis should be obligatory for individuals working at radio-relay stations.

  7. Chromosome damage and micronucleus formation in human blood lymphocytes exposed in vitro to radiofrequency radiation at a cellular telephone frequency (847.74 MHz, CDMA).

    Science.gov (United States)

    Vijayalaxmi; Bisht, K S; Pickard, W F; Meltz, M L; Roti Roti, J L; Moros, E G

    2001-10-01

    Peripheral blood samples collected from four healthy nonsmoking human volunteers were diluted with tissue culture medium and exposed in vitro for 24 h to 847.74 MHz radiofrequency (RF) radiation (continuous wave), a frequency employed for cellular telephone communications. A code division multiple access (CDMA) technology was used with a nominal net forward power of 75 W and a nominal power density of 950 W/m(2) (95 mW/cm(2)). The mean specific absorption rate (SAR) was 4.9 or 5.5 W/kg. Blood aliquots that were sham-exposed or exposed in vitro to an acute dose of 1.5 Gy of gamma radiation were included in the study as controls. The temperatures of the medium during RF-radiation and sham exposures in the Radial Transmission Line facility were controlled at 37 +/- 0.3 degrees C. Immediately after the exposures, lymphocytes were cultured at 37 +/- 1 degrees C for 48 or 72 h. The extent of genetic damage was assessed from the incidence of chromosome aberrations and micronuclei. The kinetics of cell proliferation was determined from the mitotic indices in 48-h cultures and from the incidence of binucleate cells in 72-h cultures. The data indicated no significant differences between RF-radiation-exposed and sham-exposed lymphocytes with respect to mitotic indices, frequencies of exchange aberrations, excess fragments, binucleate cells, and micronuclei. The response of gamma-irradiated lymphocytes was significantly different from that of both RF-radiation-exposed and sham-exposed cells for all of these indices. Thus there was no evidence for induction of chromosome aberrations and micronuclei in human blood lymphocytes exposed in vitro for 24 h to 847.74 MHz RF radiation (CDMA) at SARs of 4.9 or 5.5 W/kg.

  8. IgG/IgE bullous pemphigoid with CD45 lymphocytic reactivity to dermal blood vessels, nerves and eccrine sweat glands

    Directory of Open Access Journals (Sweden)

    Ana Maria Abreu-Velez

    2010-01-01

    Full Text Available Context: Bullous pemphigoid (BP, the most common autoimmune blistering disease, is mediated by autoantibodies. BP primarily affects the elderly and is characterized by the development of urticarial plaques surmounted by subepidermal blisters, and the deposition of immunoglobulins and complement at the basement membrane zone (BMZ of the skin. BP is immunologically characterized by the development of autoantibodies targeting two structural proteins of the hemidesmosomes, BP180 (collagen XVII and BP230 (BPAG1. Case Report: A 63 -year-old Caucasian female patient was evaluated for a 4 day history of several itching, erythematous blisters on her extremities. Biopsies for hematoxylin and eosin (H&E examination, as well as Periodic acid-Schiff (PAS, immunohistochemistry (IHC and direct immunofluorescence (DIF analysis were performed. Results: H&E demonstrated a subepidermal blister, with partial re-epithelialization of the blister floor. Within the blister lumen, numerous neutrophils were present, with occasional eosinophils and lymphocytes also noted. Within the dermis, a mild, superficial, perivascular and periadnexal infiltrate of lymphocytes, histiocytes and occasional eosinophils was identified, with mild perivascular leukocytoclastic debris. The PAS stain was positive at the BMZ, and around selected blood vessels, nerves and sweat glands. DIF revealed linear deposits of IgG and Complement/C3 and fibrinogen at the BMZ, and around selected dermal nerves, blood vessels and sweat glands. Strong granular deposits of IgE were also observed, colocalizing with monoclonal antibodies to Collagen IV (CIV. By IHC, positive CD45 staining of lymphocytes was seen surrounding selected dermal blood vessels, eccrine sweat glands, and nerves. Conclusion : The patient displayed IgG, IgE, and fibrinogen autoantibodies against the BMZ, as well as around some dermal nerves and sweat glands; their binding in the skin could trigger complement activation. In addition, the

  9. Melatonin may play a role in modulation of bax and bcl-2 expression levels to protect rat peripheral blood lymphocytes from gamma irradiation-induced apoptosis

    Energy Technology Data Exchange (ETDEWEB)

    Mohseni, Mehran [Department of Radiology and Medical Physics, Faculty of Paramedicine, Kashan University of Medical Sciences, Kashan (Iran, Islamic Republic of); Mihandoost, Ehsan, E-mail: mihandoost.e@gmail.com [Department of Medical Radiation Engineering, Science and Research Branch, Islamic Azad University, Tehran (Iran, Islamic Republic of); Shirazi, Alireza [Department of Medical Physics and Biomedical Engineering, Faculty of Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Sepehrizadeh, Zargham [Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Bazzaz, Javad Tavakkoly [Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Ghazi-khansari, Mahmoud [Department of Pharmacology, Faculty of Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of)

    2012-10-15

    The close relationship between free radicals effects and apoptosis process has been proved. Melatonin has been reported as a direct free radical scavenger. We investigated the capability of melatonin in the modification of radiation-induced apoptosis and apoptosis-associated upstream regulators expression in rat peripheral blood lymphocytes. Rats were irradiated with a single whole body Cobalt 60-gamma radiation dose of 8 Gy at a dose rate of 101 cGy/min with or without melatonin pretreatments at different concentrations of 10 and 100 mg/kg body weight. The rats were divided into eight groups of control, irradiation-only, vehicle-only, vehicle plus irradiation, 10 mg/kg melatonin alone, 10 mg/kg melatonin plus irradiation, 100 mg/kg melatonin alone and 100 mg/kg melatonin plus irradiation. Rats were given an intraperitoneal (IP) injection of melatonin or the same volume of vehicle alone 1 h prior to irradiation. Blood samples were taken 4, 24, 48 and 72 h after irradiation for evaluation of flow cytometric analysis of apoptotic lymphocytes using Annexin V/PI assay and measurement of bax and bcl-2 expression using quantitative real-time PCR (RT{sup 2}qPCR). Irradiation-only and vehicle plus irradiation showed an increase in the percentage of apoptotic lymphocytes significantly different from control group (P < 0.01), while melatonin pretreatments in a dose-dependent manner reduced it as compared with the irradiation-only and vehicle plus irradiation groups (P < 0.01) in all time points. This reduced apoptosis by melatonin was related to the downregulation of bax, upregulation of bcl-2, and therefore reduction of bax/bcl-2 ratio. Our results suggest that melatonin in these doses may provide modulation of bax and bcl-2 expression as well as bax/bcl-2 ratio to protect rat peripheral blood lymphocytes from gamma irradiation-induced apoptosis.

  10. Post-thaw non-cultured and post-thaw cultured equine cord blood mesenchymal stromal cells equally suppress lymphocyte proliferation in vitro.

    Directory of Open Access Journals (Sweden)

    Lynn B Williams

    Full Text Available Multipotent mesenchymal stromal cells (MSC are receiving increased attention for their non-progenitor immunomodulatory potential. Cryopreservation is commonly used for long-term storage of MSC. Post-thaw MSC proliferation is associated with a lag-phase in vitro. How this lag-phase affect MSC immunomodulatory properties is unknown. We hypothesized that in vitro there is no difference in lymphocyte suppression potential between quick-thawed cryopreserved equine cord blood (CB MSC immediately included in mixed lymphocyte reaction (MLR and same MSC allowed post-thaw culture time prior to inclusion in MLR. Cryopreserved CB-MSC from five unrelated foals were compared using two-way MLR. For each of the five unrelated MSC cultures, paired MLR assays of MSC allowed five days of post-thaw culture and MSC included in MLR assay immediately post-thawing were evaluated. We report no difference in the suppression of lymphocyte proliferation by CB-MSC that had undergone post-thaw culture and MSC not cultured post-thaw (p<0.0001. Also, there was no inter-donor variability between the lymphocyte suppressive properties of MSC harvested from the five different donors (p = 0.13. These findings suggest that cryopreserved CB-MSC may have clinical utility immediately upon thawing. One implication hereof is the possibility of using cryopreserved CB-MSC at third party locations without the need for cell culture equipment or competencies.

  11. Results of The Analysis of The Blood Beryllium Lymphocyte Proliferation Test Data From The Oak Ridge Y-12 Study

    Energy Technology Data Exchange (ETDEWEB)

    Frome, EL

    2001-12-18

    The potential hazards from exposure to beryllium or beryllium compounds in the workplace were first reported in the 1930s. The tritiated thymidine beryllium lymphocyte proliferation test (BeLPT) is an in vitro blood test that is widely used to screen beryllium exposed workers in the nuclear industry for sensitivity to beryllium. Newman [18] has discussed the clinical significance of the BeLPT and described a standard protocol that was developed in the late 1980s. Cell proliferation is measured by the incorporation of tritiated thymidine into dividing cells on two culture dates and using three concentrations of beryllium sulfate. Results are expressed as a ''stimulation index'' (SI) which is the ratio of the amount of tritiated thymidine (measured by beta counts) in the stimulated cells divided by the counts for the unstimulated cells on the same culture day. Several statistical methods for use in the routine analysis of the BeLPT were considered in the early 1990's by Frome et al. [7]. The least absolute values (LAV) method was recommended for routine analysis of the BeLPT. The purposes of this report are to further evaluate the LAV method using new data, and to describe a new method for identification of an abnormal or borderline test. This new statistical biological positive (SBP) method reflects the clinical judgment that (1) at least two SIs show a ''positive'' response to beryllium, and (2), that the maximum of the six SIs must exceed a cut point that is determined from a reference data set of normal individuals whose blood has been tested by the same method in the same serum. The new data is from the Y-12 facility in Oak Ridge and consist of 1080 worker and 33 nonexposed control BeLPTs (all tested in the same serum). Graphical results are presented to explain the statistical method, and the new SBP method is applied to the Y-12 group. The true positive rate and specificity of the new method were estimated to be 86

  12. Apoptosis of granulocytes and lymphocytes in peripheral blood in rabbits infected with haemagglutinating and non-haemagglutinating antigenic variants of the RHD (rabbit haemorrhagic disease) virus.

    Science.gov (United States)

    Niedźwiedzka-Rystwej, P; Tokarz-Deptuła, B; Deptuła, W

    2013-01-01

    This paper attempts to study the dynamics of apoptosis of granulocytes and lymphocytes in peripheral blood in rabbits infected with haemagglutinating (Vt97, Triptis, Hartmannsdorf) and non-haemagglutinating (Pv97, 9905 RHDVa) antigenic variants of the RHD virus. The pathogenicity of those antigenic variants was also assessed by recording the mortality of the infected animals. The animals were infected with antigenic variants and blood was sampled at hour 0, 4, 8, 12, 24, 36 p.i. and the percentage of apoptotic granulocytes and lymphocytes was measured with the use of flow cytometry. The results of the study showed that apoptosis is included during RHDV infection, as the number of apoptotic granulocytes and lymphocytes increases throughout the experiment; depending on the antigenic variant, apoptosis joins in at 4-8-12 h p.i. and lasts until 24-36 h p.i. Furthermore, the mortality of rabbits infected with the examined strains of RHD virus varied from 30% to 100%. This study performed for the first time in this manner, indicates the importance of apoptosis during infection with the RHD virus.

  13. [Effect of accelerated heavy ions of carbon 12C, neon 20Ne and iron 56Fe on the chromosomal apparatus of human blood lymphocytes in vitro].

    Science.gov (United States)

    Repina, L A

    2011-01-01

    Cytogenetic assay of the chromosomal apparatus of human blood lymphocytes was carried out after in vitro irradiation by heavy charged particles with high LET values. Blood plasm samples enriched with lymphocytes were irradiated by accelerated ions of carbon 12C (290 MeV/nucleon and LET = 70 keV/microm), neon 20Ne (400 MeV/nucleon and LET = 70 keV/microm), and iron 56Fe (500 MeV/nucleon and LET = 200 keV/microm) in the dose range from 0.25 to 1 Gy. Rate of chromosome aberrations showed a linear dependence on doses from the densely ionizing radiations with high LET values. Frequency of dicentrics and centric rings in human lymphocytes irradiated by 12C with the energy of 290 MeV/nucleon was maximal at 1 Gy (p < 0.05) relative to the other heavy particles. It was found that relative biological effectiveness of heavy nuclei is several times higher than of 60Co gamma-radiation throughout the range of doses in this investigation.

  14. Radio-induced apoptosis of peripheral blood CD8 T lymphocytes is a novel prognostic factor for survival in cervical carcinoma patients

    Energy Technology Data Exchange (ETDEWEB)

    Ordonez, R.; Federico, M. [Hospital Universitario de Gran Canaria Dr. Negrin, Radiation Oncology Department, Las Palmas de Gran Canaria (Spain); Henriquez-Hernandez, L.A.; Pinar, B.; Lloret, M.; Lara, P.C. [Hospital Universitario de Gran Canaria Dr. Negrin, Radiation Oncology Department, Las Palmas de Gran Canaria (Spain); Universidad de Las Palmas de Gran Canaria, Clinical Sciences Department, Las Palmas de Gran Canaria (Spain); Instituto Canario de Investigacion del Cancer (ICIC), Santa Cruz de Tenerife (Spain); Valenciano, A. [Instituto Canario de Investigacion del Cancer (ICIC), Santa Cruz de Tenerife (Spain); Bordon, E. [Universidad de Las Palmas de Gran Canaria, Clinical Sciences Department, Las Palmas de Gran Canaria (Spain); Rodriguez-Gallego, C. [Hospital Universitario de Gran Canaria Dr. Negrin, Immunology Department, Las Palmas de Gran Canaria (Spain)

    2014-02-15

    A close relationship exists between immune response and tumor behavior. This study aimed to explore the associations between radiation-induced apoptosis (RIA) in peripheral blood lymphocytes (PBL) and clinical pathological variables. Furthermore, it assessed the role of RIA as a prognostic factor for survival in cervical carcinoma patients. Between February 1998 and October 2003, 58 consecutive patients with nonmetastatic, localized stage I-II cervical carcinoma who had been treated with radiotherapy (RT) ± chemotherapy were included in this study. Follow-up ended in January 2013. PBL subpopulations were isolated and irradiated with 0, 1, 2 and 8 Gy then incubated for 24, 48 and 72 h. Apoptosis was measured by flow cytometry and the ss value, a parameter defining RIA of lymphocytes, was calculated. Mean follow-up duration was 111.92 ± 40.31 months. Patients with lower CD8 T lymphocyte ss values were at a higher risk of local relapse: Exp(B) = 5.137, confidence interval (CI) 95 % = 1.044-25.268, p = 0.044. Similar results were observed for regional relapse: Exp(B) = 8.008, CI 95 % = 1.702-37.679, p = 0.008 and disease relapse: Exp(B) = 6.766, CI 95 % = 1.889-24.238, p = 0.003. In multivariate analysis, only the CD8 T lymphocyte ss values were found to be of prognostic significance for local disease-free survival (LDFS, p = 0.049), regional disease-free survival (RDFS, p = 0.002), metastasis-free survival (MFS, p = 0.042), disease-free survival (DFS, p = 0.001) and cause-specific survival (CSS p = 0.028). For the first time, RIA in CD8 T lymphocytes was demonstrated to be a predictive factor for survival in cervical carcinoma patients. (orig.)

  15. Analysis of T-lymphocyte subtypes of the peripheral blood and skin delayed-type hypersensitivity in patients with hyper-IgE syndrome

    Institute of Scientific and Technical Information of China (English)

    Xiaobing Lei; Shengshun Tan; Weihui Zeng; Junmin Wang; Panjian Zhang; Yuan Yuan

    2005-01-01

    Objective: To study the function of cellular immunity in patients with hyper-IgE syndrome (HIE). Methods:T-lymphocyte subtypes of the peripheral blood and cutaneous delayed-type hypersensitivity (DTH) response to two recall antigens, tetanus toxoid (TT) and purified protein derivative(PPD), were measured in five patients with HIE and 15 healthy controls, respectively. Results: The CD4+ cell counts in HIE group were significantly lower than those in control group ( P < 0.01). In contrast, CD8 + cells were significantly higher than those in the control ( P < 0.01). The induration sizes of DTH response to two recall antigens were smaller in HIE group than those in the control group ( P < 0.01 ). Conclusion: There was an immunologic dysfunction of T lymphocytes in the patients with HIE.

  16. Intratumoural and peripheral blood lymphocyte subsets in patients with metastatic renal cell carcinoma undergoing interleukin-2 based immunotherapy: association to objective response and survival

    DEFF Research Database (Denmark)

    Donskov, F; Bennedsgaard, K M; Von Der Maase, H

    2002-01-01

    The aim of the present study was to analyse lymphocyte subsets in consecutive peripheral blood samples and consecutive tumour tissue core needle biopsies performed before and during interleukin-2 based immunotherapy, and to correlate the findings with objective response and survival. Twenty...... response or survival. Within the tumour tissue at baseline, a significant positive correlation between CD4 (P=0.027), CD8 (P=0.028), CD57 (P=0.007) and objective response was demonstrated. After one month of immunotherapy, a significant positive correlation between intratumoral CD3 (P=0.026), CD8 (P=0...... of lymphocyte subsets in the tumour reduction in responding patients during interleukin-2 based immunotherapy. Confirmation of the results requires further studies including a larger number of patients....

  17. Defining Molecular Phenotypes of Mesenchymal and hematopoietic Stem Cells derived from Peripheral blood of Acute Lymphocytic Leukemia patients for regenerative stem cell therapy.

    Science.gov (United States)

    Potdar, Pd; Subedi, Rp

    2011-01-01

    Acute Lymphocytic Leukemia (ALL) is a clonal myeloid disorder affecting all age groups, characterized by accumulation of immature blast cells in bone marrow and in peripheral blood. Autologous Bone Marrow Transplantation is a present treatment for cure of ALL patients, which is very expensive, invasive process and may have possibility of transplantation of malignant stem cells to patients. In the present study, we hypothesized to isolate large number of normal Mesenchymal & Hematopoietic stem cells from peripheral blood of ALL patients, which will be further characterized for their normal phenotypes by using specific molecular stem cell markers. This is the first study, which defines the existing phenotypes of isolated MSCs and HSCs from peripheral blood of ALL patients. We have established three cell lines in which two were Mesenchymal stem cells designated as MSCALL and MSCnsALL and one was suspension cell line designated as HSCALL. The HSCALL cell line was developed from the lymphocyte like cells secreted by MSCALL cells. Our study also showed that MSCALL from peripheral blood of ALL patient secreted hematopoietic stem cells in vitro culture. We have characterized all three-cell lines by 14 specific stem cell molecular markers. It was found that both MSC cell lines expressed CD105, CD13, and CD73 with mixed expression of CD34 and CD45 at early passage whereas, HSCALL cell line expressed prominent feature of hematopoietic stem cells such as CD34 and CD45 with mild expression of CD105 and CD13. All three-cell lines expressed LIF, OCT4, NANOG, SOX2, IL6, and DAPK. These cells mildly expressed COX2 and did not express BCR-ABL. Overall it was shown that isolated MSCs and HSCs can be use as a model system to study the mechanism of leukemia at stem cell level and their use in stem cell regeneration therapy for Acute Lymphocytic Leukemia.

  18. Hydrophobic sodium fluoride-based nanocrystals doped with lanthanide ions: assessment of in vitro toxicity to human blood lymphocytes and phagocytes.

    Science.gov (United States)

    Sojka, Bartlomiej; Kuricova, Miroslava; Liskova, Aurelia; Bartusova, Maria; Banski, Mateusz; Misiewicz, Jan; Dusinska, Maria; Horvathova, Mira; Jahnova, Eva; Ilavska, Silvia; Szabova, Michaela; Rollerova, Eva; Podhorodecki, Artur; Tulinska, Jana

    2014-11-01

    In vitro immunotoxicity of hydrophobic sodium fluoride-based nanocrystals (NCs) doped with lanthanide ions was examined in this study. Although there is already a significant amount of optical and structural data on NaYF4 NCs, data on safety assessment are missing. Therefore, peripheral whole blood from human volunteers was used to evaluate the effect of 25 and 30 nm hydrophobic NaYF4 NCs dissolved in cyclohexane (CH) on lymphocytes, and of 10 nm NaYF4 NCs on phagocytes. In the concentration range 0.12-75 µg cm(-2) (0.17-106 µg ml(-1) ), both 25 and 30nm NaYF4 NCs did not induce cytotoxicity when measured as incorporation of [(3) H]-thymidine into DNA. Assessment of lymphocyte function showed significant suppression of the proliferative activity of T-lymphocytes and T-dependent B-cell response in peripheral blood cultures (n = 7) stimulated in vitro with mitogens phytohemagglutinin (PHA) and pokeweed (PWM) (PHA > PWM). No clear dose-response effect was observed. Phagocytic activity and respiratory burst of leukocytes (n = 5-8) were generally less affected. A dose-dependent suppression of phagocytic activity of granulocytes in cultures treated with 25 nm NCs was observed (vs. medium control). A decrease in phagocytic activity of monocytes was found in cells exposed to higher doses of 10 and 30 nm NCs. The respiratory burst of phagocytes was significantly decreased by exposure to the middle dose of 30 nm NCs only. In conclusion, our results demonstrate immunotoxic effects of hydrophobic NaYF4 NCs doped with lanthanide ions to lymphocytes and to lesser extent to phagocytes. Further research needs to be done, particularly faze transfer of hydrophobic NCs to hydrophilic ones, to eliminate the solvent effect.

  19. Blood smear

    Science.gov (United States)

    ... some red blood cells shaped like spheres ( hereditary spherocytosis ) Increased breakdown of RBCs Presence of RBCs with ... normal Red blood cells, elliptocytosis Red blood cells, spherocytosis Acute lymphocytic leukemia - photomicrograph Red blood cells, multiple ...

  20. A clinical observation of the influence of deep brain stimulation on peripheral blood lymphocytes in patients with Parkinson's disease

    Directory of Open Access Journals (Sweden)

    LIU Jing

    2013-07-01

    Full Text Available Objective To study the changing in number of peripheral blood lymphocyte (PBL of patients with Parkinson's disease (PD after deep brain stimulation (DBS, and to explore the mechanism of DBS in treating PD. Methods One hundred and thirty PD patients were divided into 2 groups, namely, non-DBS group [N = 105; 68 males and 37 females; mean age (61.54 ± 10.44 years; mean duration (7.29 ± 4.57 years], and DBS group [N = 25; 16 males and 9 females; mean age (59.20 ± 10.67 years; mean disease duration (12.16 ± 4.79 years]. There were 73 healthy subjects [37 males and 36 females; mean age (61.89 ± 12.20 years] in control group. The differences of the number of PBL among the 3 groups were analyzed. Spearman's rank correlation analysis was used to assess the relationship between PBL number and influenzing factors [gender, age, disease duration, Unified Parkinson's Disease Rating Scale (UPDRS Ⅲ score, Hoehn-Yahr (H-Y stage, and drug equivalent daily dose]. Results The number of PBL in non-DBS group was less than that in control group (P = 0.000. There was significant correlation between UPDRS Ⅲ and PBL number (rs = - 0.403, P = 0.031. No correlation was found between PBL number and gender, age, disease duration, H-Y stage or drug equivalent daily dose (P > 0.05, for all. No difference was shown between PBL number in control group and in DBS group (P = 0.137 and no correlations were found with clinical variables (P > 0.05. The PBL number in non-DBS group was less than that in DBS group (P = 0.006. With the same H-Y stage, PBL number in non-DBS group was also less than that in DBS group in Mann-Whitney U test (H-Y 2.5: Z = - 2.197, P = 0.043; H-Y 3: Z = - 1.875, P = 0.027; H-Y 4: Z = - 3.760, P = 0.016. Conclusion The changing in the number of PBL is the specific feature of PD and may be correlated with the immuno-inflammation of central nervous system, which may be relieved by DBS.

  1. Autoimmune Demyelinating Polyneuropathy as a Manifestation of Chronic Graft-versus-Host Disease after Adult Cord Blood Transplantation in a Patient with Chronic Lymphocytic Leukemia

    Directory of Open Access Journals (Sweden)

    Fredrick Hogan

    2014-01-01

    Full Text Available Immune mediated demyelinating disease after allogeneic stem cell transplantation is a rare entity with unclear etiology. Acute inflammatory demyelinating polyneuropathy (AIDP has been reported after related and adult unrelated allogeneic stem cell transplantation but no such case has been reported after unrelated cord blood transplantation. We hereby present the first case of AIDP after double umbilical cord blood transplantation (DUCBT. A 55-year-old man with chronic lymphocytic leukemia (CLL received a cord blood transplant for relapsed refractory disease with high risk cytogenetics. On day 221, patient presented with skin rash, tingling in both lower extremites, and ascending paralysis that progressed rapidly over the course of 2 days. The workup resulted in a diagnosis of AIDP and administration of intravenous immunoglobulins plus steroids was initiated. Motor and sensory powers were fully recovered and his chronic GVHD was managed for several months with single agent sirolimus.

  2. 焦炉工人外周血淋巴细胞凋亡特征分析%Characteristics of Peripheral Blood Lymphocytes Apoptosis in Coke-oven Workers

    Institute of Scientific and Technical Information of China (English)

    胡志鹏; 张红明; 李卫星; 张晶; 宋秋坤; 王保平; 李治玉; 郑金平

    2011-01-01

    [目的]研究焦炉工人外周血淋巴细胞凋亡特征及其影响因素,为加强焦炉工的健康防护提供指导依据.[方法]收集343名焦炉工人班后尿,高效液相色谱法测定尿1-羟基芘(1-OHP)浓度,按尿1-OHP浓度将研究对象分为低、中、高暴露组.分光光度法测定血浆乳酸脱氢酶(LDH)活性和丙二醛(MDA焓量,流式细胞仪测定淋巴细胞凋亡率.[结果]以低暴露组淋巴细胞凋亡率95%上限值(7.90%)为界值判断淋巴细胞凋亡阳性,结果显示,低、中、高暴露组淋巴细胞凋亡阳性率随剂量升高而升高(趋势检验P<O.01).经多因素Logistic回归分析,中暴露组、高暴露组和夜班是淋巴细胞凋亡阳性的主要危险因素(OR分别为3.42、8.58和5.05,P值均小于0.05 o相关分析结果显示,淋巴细胞凋亡率与尿1-OHP含量呈正相关(r=0.113,P<0.05).[结论]焦炉逸散物暴露可导致部分焦炉工人淋巴细胞凋亡率升高,且与尿1-OHP水平有关.夜班是主要影响因素.%[Objective]To investigate the characteristics of lymphocytes apoptosis in peripheral blood of coke-oven workers and its influential factors.[Methods]In this study, a total of 343 coke-oven workers were included.1-Hydroxypyrene ( 1-OHP ) in worker's post-shift urine sample was determined by high performance liquid chromatography.Subjects were divided into low, medium or high exposure group according to the 1-OHP concentration.Lactic dehydrogenase ( LDH )activity and malondialehehyche( MDA )concentration in plasma were determined using speetrophotometry.Lymphocyte apoptosis in peripheral blood was determined using flow cytometry.[Results]The upper bound lymphocyte apoptosis rates of 95% subjects in the low exposure group ( 7.9% ) was defined as the threshold to determine positive lymphocyte apoptosis.The positive rate of lymphocyte apoptosis in the low, medium and high exposure groups were increased with the exposure levels( chi-square trend test, P< 0

  3. Relationships between in vitro lymphoproliferative responses and levels of contaminants in blood of free-ranging adult harbour seals (Phoca vitulina) from the North Sea

    Energy Technology Data Exchange (ETDEWEB)

    Dupont, Aurélie [Laboratory of Oceanology – MARE Center, University of Liège, B6c, allée de la chimie 3, B-4000 Liège (Sart-Tilman) (Belgium); Siebert, Ursula [Institute for Terrestrial and Aquatic Wildlife Research, University of Veterinary Medicine Hannover, Foundation, Werftstrasse 6, 25761 Buesum (Germany); Covaci, Adrian; Weijs, Liesbeth [Toxicological Centre, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk (Belgium); Systemic Physiological and Ecotoxicological Research (SPHERE), Department of Biology, University of Antwerp, Groenenborgerlaan 171, 2020 Antwerp (Belgium); Eppe, Gauthier [Center for Analytical Research and Technology (CART), Inorganic Analytical Chemistry, University of Liège, B6c, allée de la chimie 3, B-4000 Liège (Sart-Tilman) (Belgium); Debier, Cathy [Unit of Nutrition Biochemistry, Catholic University of Leuven, Place Croix du Sud 2/8, 1348 Louvain-la-Neuve (Belgium); De Pauw-Gillet, Marie-Claire [Mammalian Cell Culture Laboratory (GIGA-R), University of Liège, B6c, allée de la chimie 3, B-4000 Liège (Sart-Tilman) (Belgium); Das, Krishna, E-mail: Krishna.das@ulg.ac.be [Laboratory of Oceanology – MARE Center, University of Liège, B6c, allée de la chimie 3, B-4000 Liège (Sart-Tilman) (Belgium)

    2013-10-15

    Highlights: •Blood samples were taken from adult harbour seals in good body condition. •Trace element and POP levels, as well as haematological parameters were determined. •Peripheral blood leucocytes were isolated from the same blood samples. •The in vitro lymphocyte stimulation indices were calculated. •No clear correlation between the lymphocyte stimulation indices and pollutants could be highlighted. -- Abstract: In vitro culture of peripheral blood leucocytes (PBLs) is currently used in toxicological studies of marine mammals. However, blood cells of wild individuals are exposed in vivo to environmental contaminants before being isolated and exposed to contaminants in vitro. The aim of this study was to highlight potential relationships between blood contaminant levels and in vitro peripheral blood lymphocyte proliferation in free-ranging adult harbour seals (Phoca vitulina) from the North Sea. Blood samples of 18 individuals were analyzed for trace elements (Fe, Zn, Se, Cu, Hg, Pb, Cd) and persistent organic contaminants and metabolites (ΣPCBs, ΣHO-PCBs, ΣPBDEs, 2-MeO-BDE68 and 6-MeO-BDE47, ΣDDXs, hexachlorobenzene, oxychlordane, trans-nonachlor, pentachlorophenol and tribromoanisole). The same samples were used to determine the haematology profiles, cell numbers and viability, as well as the in vitro ConA-induced lymphocyte proliferation expressed as a stimulation index (SI). Correlation tests (Bravais-Pearson) and Principal Component Analysis with multiple regression revealed no statistically significant relationship between the lymphocyte SI and the contaminants studied. However, the number of lymphocytes per millilitre of whole blood appeared to be negatively correlated to pentachlorophenol (r = −0.63, p = 0.005). In adult harbour seals, the interindividual variations of in vitro lymphocyte proliferation did not appear to be directly linked to pollutant levels present in the blood, and it is likely that other factors such as age, life

  4. [Changes of content of regulatory lymphocytes and concentration of soluble interleukine-2 receptor in blood of patients with ischemic heart disease after coronary artery angioplasty with implantation of stents with rapamycin covering].

    Science.gov (United States)

    Potekhina, A V; Sokolov, V O; Pylaeva, E A; Provatorov, S I; Masenko, V P; Bosykh, E G; Noeva, E A; Krasnikova, T L; Aref'eva, T I

    2011-01-01

    We studied dynamics of content of subpopulation of lymphocytes including regulatory and effector T-lymphocytes as well as concentration of soluble form of interleukine-2 receptor (sCD25) in peripheral blood of patients after coronary stenting (CS) with implantation of stents with rapamycin covering (SRC). We included into the study 62 patients with stable effort II-III functional class angina. Coronary angiography (CA) was carried out in all, CS with implantation of 1 - 2 SRC - in 42 patients. Blood samples were taken before CA/CS, in 24, 48 hours, 7 days, 1 and 3 months after intervention. Content of T-, helper and cytotoxic T-cells, -, NK-, NKT-cells, activated effector T-lymphocytes (CD4+CD251owCD127high) and regulatory T-lymphocytes (CD4+CD25highCD1271ow) were measured by direct immunofluorescence and flow cytometry. CD4+ lymphocytes were isolated from mononuclear cell fraction of donor blood by magnetic separation. Content of regulatory T-lymphocytes in culture were determined by expression of a specific marker FOXP3+. Concentration of sCD25 was measured by chemiluminescent method. It was shown that content of main subpopulations of lymphocytes in blood changed after CS or CF. Blood content of regulatory T-lymphocytes and sCD25 significantly increased after 7 days and 1 month after CS but not after CA. Plasma sCD25 concentration correlated with content of regulatory T-lymphocytes in 1 month after SRC implantation. During cultivation of CD4+ lymphocytes in the presence of rapamycin we noted antiproliferative effect relative to FOXP3-cells and accumulation of regulatory +-lymphocytes. Thus implantation of SRC in coronary arteries leads to increase of number of circulating regulatory T-lymphocytes and blood concentration of sCD25. Changes of these parameters after CS can reflect peculiarities of local and systemic reaction arising in response to introduction of stent with drug covering and be significant for assessment of prognosis of the disease.

  5. Unstable chromosome aberrations on peripheral blood lymphocytes from patients with cervical uterine cancer following radiotherapy; Aberracoes cromossomicas instaveis em linfocitos de pacientes com cancer de colo de utero

    Energy Technology Data Exchange (ETDEWEB)

    Magnata, Simey de Souza Leao Pereira

    2002-09-01

    Absorbed dose determination is an important step for risk assessment related to an exposure to ionizing radiation. However, physical dosimetry cannot be always performed, principally in the case of retrospective estimates. In this context, the use of bioindicators (biological effects) has been proposed, which defines the so-called biological dosimetry. In particular, scoring of unstable chromosomes aberrations (dicentrics, centric rings and fragments) of peripheral blood lymphocytes, while is the most reliable biological method for estimating individual exposure to ionizing radiation. In this work, blood samples from 5 patients, with cervical uterine cancer, were evaluated after partial-body radiotherapy with a source of {sup 69} Co. For this, conventional cytogenetic method was employed, based on Giemsa coloration and fluorescence in situ hybridization, in order to correlate the frequency of unstable chromosome aberrations of blood lymphocytes with absorbed dose, as a result of the radiotherapy. A good agreement was observed between the frequency of chromosome aberrations scored and the values of dose previously calculated by physical dosimetry during patient's radiotherapy. The results presented in this work point out the importance of concerning analyses of unstable chromosome aberrations as biological dosimeter in the investigation of partial-body exposure to ionizing radiation. (author)

  6. Properties of Na+ ,K+ -activated, Mg2+ -dependent ATP-hydrolyze of blood lymphocytes in patients with reаctive arthritis

    Directory of Open Access Journals (Sweden)

    O.V. Melnyk

    2013-11-01

    with RеA. We observed that Na+,K+-ATPase of peripheral blood lymphocytes of patients with RеA retains its endogenous receptor properties – sensitivity to ouabain does not change. It is assumed that under conditions of rheumatic pathology the impact on the Na+,K+-ATPase structure occurs both externally and on the cytoplasmic membrane surface. The above experimental data can be used for further clarification of the membrane mechanisms of ion exchange in immunocompetent cells of patients suffering from autoimmune diseases.

  7. Determination of CD30 Expression on Peripheral Blood T Lymphocyte Subsets in Patients with Hemorrhagic Fever with Renal Syndrome by FCM

    Institute of Scientific and Technical Information of China (English)

    XIONG; Lijuan; LUO; Duande; ZENG; Linglan; LI; Shuli

    2001-01-01

    To determine the CD30 expression on peripheral blood T lymphocyte subsets in patients with hemorrhagic fever with renal syndrome (HFRS) and its clinical implications, double immunofluorescence technique and flow cytometry were used. There was no significant difference among the severe group, mild-moderate group and normal control group in the CD4+CD30- T lymphocyte subset.While the CD4+CD30+ T cells of HFRS patients were increased and the difference between severe group and mild-moderate group or normal control group were very significant (P<0. 01) and the difference between the mild-moderate group and normal control group was also significant (P<0. 05). The CD8+CD30-T cells were increased while the CD8+CD30+ T cells decreased obviously in HFRS patients,and the differences among three groups in both subsets were very significant (P<0. 01). The results showed that the humoral immunity and cellular immunity are overactive in HFRS patients during acute phase. The loss of balance between T lymphocyte subsets may play an important role in the pathophysiology of HFRS and is closely correlated with the severity of the HFRS.

  8. Exposure to brominated trihalomethanes in water during pregnancy and micronuclei frequency in maternal and cord blood lymphocytes

    DEFF Research Database (Denmark)

    Stayner, Leslie Thomas; Pedersen, Marie; Patelarou, Evridiki;

    2014-01-01

    BACKGROUND: Water disinfection by-products have been associated with an increased cancer risk. Micronuclei (MN) frequency in lymphocytes is a marker of genomic damage and can predict adult cancer risk. OBJECTIVE: We evaluated maternal exposure to drinking water brominated trihalomethanes (BTHM) i...

  9. Corticosteroids decrease the expression of beta 2-microglobulin and histocompatibility antigens on human peripheral blood lymphocytes in vitro

    DEFF Research Database (Denmark)

    Hokland, M; Larsen, B; Heron, I

    1982-01-01

    . Both antigens were found to be decreased, dexamethasone typically in a concentration of 10-6 mol/l causing a decrease in surface beta 2-microglobulin of 15% after an incubation period of 24 hr. The expression of two other lymphocyte surface antigens, Igm and Thy antigens, measured in parallel with beta...

  10. Blood concentrations of hydroxychloroquine and its desethyl derivative correlate negatively with the percentage of CD45RO+ cells among CD4+ lymphocytes in hydroxychloroquine-treated lupus patients.

    Science.gov (United States)

    Sailler, Laurent; Puissant, Bénédicte; Méliani, Pascal; Castex, Jean Olivier; Saivin, Sylvie; Adoue, D; Fournie, B; Arlet, Philippe; Montastruc, Jean-Louis; Lapeyre-Mestre, Maryse; Pourrat, Jacques; Blancher, Antoine

    2007-06-01

    The objective of the study was to investigate the influence of the blood concentrations of hydroxychloroquine ([HCQ]) and its derivative desethylhydroxychloroquine ([DHCQ]) on lymphocyte activation or differentiation in HCQ-treated lupus patients. We studied the correlations between [HCQ], [DHCQ], and the frequency of various lymphocyte subsets in 58 HCQ-treated lupus patients (mean HCQ dose: 4.93 +/- 1.58 mg/kg/day; mean duration of the disease: 122 +/- 64 months). [HCQ] and [DHCQ] were determined by high-performance liquid chromatography (HPLC). Lymphocyte markers were studied by flow cytometry using monoclonal anti-CD3, -CD4, -CD8, -CD25, -DR, -CD45RA, -CD45RO, -CD19, -CD38, and -CD86 antibodies. sIL2-R serum concentrations were measured by enzyme-linked immunosorbent assay (ELISA). [HCQ] and [DHCQ] were 599.9 ng/mL (median: 529.5; range: 55-1935) and 353.43 (median: 286 ng/mL; range: 118-1090). In a multiple regression analysis, [HCQ] and [DHCQ] were associated with the HCQ prescribed dose in mg/kg/day (P = 0.0002 and P = 0.03) and with compliance to the treatment (P = 0.004 and P = 0.03). We found a negative correlation between [HCQ], [DHCQ], and the CD45RO+ cell frequency among CD3+CD4+ cells (P = 0.03 and P = 0.007, respectively). Other lymphocyte subset markers (LSMs) and sIL2-R concentrations were not significantly associated with [HCQ] or [DHCQ]. In the multiple regression analysis, CD45RO+ expression was negatively influenced by [HCQ] (P = 0.005), and positively influenced by smoking habits (P = 0.005) and age (P = 0.005). Similar results were found in the multivariate model including [DHCQ]. Disease activity and taking more than 10 mg/day of corticosteroids or an immunosuppressive drug did not influence CD45RO+ expression. Lupus patients had less CD3+CD4+CD45RO+ cells than controls (P = 0.03). In lupus patients, HCQ and DHCQ may alter the generation or the blood circulation of CD4+CD45RO+ lymphocytes in a concentration-dependent pattern.

  11. HIV-1 infection and first line ART induced differential responses in mitochondria from blood lymphocytes and monocytes: the ANRS EP45 "Aging" study.

    Directory of Open Access Journals (Sweden)

    Sophie Perrin

    Full Text Available BACKGROUND: The ANRS EP45 "Aging" study investigates the cellular mechanisms involved in the accelerated aging of HIV-1 infected and treated patients. The data reported focus on mitochondria, organelles known to be involved in cell senescence. METHODS: 49 HIV-1 infected patients untreated with antiretroviral therapy, together with 49 seronegative age- and sex-matched control subjects and 81 HIV-1 infected and treated patients, were recruited by 3 AIDS centres (Marseille, Montpellier, Nice; France; http://clinicaltrials.gov/, NCT01038999. In more than 88% of treated patients, the viral load was 500/mm(3. ROS (reactive oxygen species production and ΔΨm (inner membrane potential were measured by flow cytometry in blood lymphocytes and monocytes (functional parameters. Three mitochondrial network quantitative morphological parameters were computed using confocal microscopy and image analysis. Three PBMC mitochondrial proteins (porin and subunits 2 and 4 of cytochrome C oxidase encoded by mtDNA or nuclear DNA, respectively were analysed by western blotting. RESULTS: Quantitative changes in PBMC mitochondrial proteins were not induced by either HIV-1 infection or ART. Discriminant analysis integrating functional (ROS production and ΔΨm or morphological (network volume density, fragmentation and branching parameters revealed HIV-1 infection and ART differential effects according to cell type. First line ART tended to rescue lymphocyte mitochondrial parameters altered by viral infection, but induced slight changes in monocytes. No statistical difference was found between the effects of three ART regimens on mitochondrial parameters. Correlations between functional parameters and viral load confirmed the damaging effects of HIV-1 in lymphocyte mitochondria. CONCLUSIONS: In patients considered to be clinically stable, mitochondria exhibited functional and morphological modifications in PBMCs resulting from either direct or indirect effects of HIV-1

  12. Expression of IL-2R on Peripheral Blood Lymphocytes of Patients with Colorectal Cancer and Its Clinical Significance

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective: To study expression of membrane receptors ofinterleukin-2 (CD25) on the peripheral blood lymphocytes (PBL) of patients with colorectal cancer and its clinical significance. Methods: CD25 percentages (CD25%) in PBL of 105 colorectal cancer patients before operation and 100 normal individuals were examined by flow cytometer, and the results were clinically and pathologically analyzed. Results: The mean of CD25% in PBL of the normal individuals was 17.24± 5.33, it was significantly lower (p<0.01) than that of the colon cancer patients (21.29± 7.95) or rectal cancer patients (21.62± 6.11). In contrast to the normal individuals, the means of CD25% in PBL in ulcer type (20.53± 6.50) or protruded type (21.56± 6.16) colorectal cancer patients were notably elevated (P<0.01). The significant difference (P<0.01) of means of CD25% in PBL was observed between the normal individuals and patients with less than 4 cm mass (22.10± 5.43) or 4cm- 8cm mass (20.90± 6.96). The significant difference (P<0.05) of means of CD25% in PBL was also observed between the normal individuals and patients with greater than 8 cm mass (21.56± 5.41). The mean of CD25% in PBL in patients with well differentiation colorectal cancer was 22.20± 5.50, it was significantly higher than that in normal individuals (P<0.05). The means of CD25% in PBL in patients with middle or poor differentiation colorectal cancer were 21.30± 6.89 and 22.15± 5.71 respectively, they were obviously higher than that in normal individuals (P<0.01). The significant difference (P<0.01) of means of CD25% in PBL was present between the colorectal cancer patients without metastatic lymph nodes (22.06± 6.90) and normal individuals. The significant difference (P<0.05) of means of CD25% in PBL was present between the colorectal cancer patients with metastatic lymph nodes (20.73± 6.40) and normal individuals. The means of CD25% in PBL in colorectal cancer patients in various clinic stages were significantly higher

  13. A sportomics strategy to analyze the ability of arginine to modulate both ammonia and lymphocyte levels in blood after high-intensity exercise

    Directory of Open Access Journals (Sweden)

    Gonçalves Luis

    2012-06-01

    Full Text Available Abstract Background Exercise is an excellent tool to study the interactions between metabolic stress and the immune system. Specifically, high-intensity exercises both produce transient hyperammonemia and influence the distribution of white blood cells. Carbohydrates and glutamine and arginine supplementation were previously shown to effectively modulate ammonia levels during exercise. In this study, we used a short-duration, high-intensity exercise together with a low carbohydrate diet to induce a hyperammonemia state and better understand how arginine influences both ammonemia and the distribution of leukocytes in the blood. Methods Brazilian Jiu-Jitsu practitioners (men, n = 39 volunteered for this study. The subjects followed a low-carbohydrate diet for four days before the trials and received either arginine supplementation (100 mg·kg-1 of body mass·day-1 or a placebo. The intergroup statistical significance was calculated by a one-way analysis of variance, followed by Student’s t-test. The data correlations were calculated using Pearson’s test. Results In the control group, ammonemia increased during matches at almost twice the rate of the arginine group (25 mmol·L-1·min-1 and 13 μmol·L-1·min-1, respectively. Exercise induced an increase in leukocytes of approximately 75%. An even greater difference was observed in the lymphocyte count, which increased 2.2-fold in the control group; this increase was partially prevented by arginine supplementation. The shape of the ammonemia curve suggests that arginine helps prevent increases in ammonia levels. Conclusions These data indicate that increases in lymphocytes and ammonia are simultaneously reduced by arginine supplementation. We propose that increased serum lymphocytes could be related to changes in ammonemia and ammonia metabolism.

  14. [EFFECT OF 4-METHYLPYRAZOLE ON IMMUNE RESPONSE, FUNCTION OF Th1 AND Th2 LYMPHOCYTES, AND CYTOKINE CONCENTRATION IN RAT BLOOD AFTER ACUTE METHANOL POISONING].

    Science.gov (United States)

    Zabrodskii, P F; Maslyakov, V V; Gromov, M S

    2016-01-01

    It was established in experiments on noninbred albino rats that the acute intoxication with methanol (1.0 LD50) decreased cellular and humoral immune responses, Th2-lymphocyte activity (to a greater extent as compared to the function of Th1 cells), reduced the blood concentration of immunoregulatory (IFN-g, IL-2, IL-4) and proinflammatory (TNF, IL-1b, IL-6) cytokines on the average by 36.5% (p Methanol antidote 4-methylpyrazole (non-competitive inhibitor of alcohol dehydrogenase) administered upon acute intoxication with methanol at a dose of 1.0 DL50 partially reduces the intoxication-induced suppression of humoral and cellular immune response, activity of T-helper cells, and production of IL-4 and restores blood levels of TNF, IL-1b, IFN-γ, IL-4, IL-2, IL-6 to the control values.

  15. High Viral Loads of Epstein-Barr Virus DNA in Peripheral Blood of Patients with Chronic Lymphocytic Leukemia Associated with Unfavorable Prognosis.

    Science.gov (United States)

    Grywalska, Ewelina; Roliński, Jacek; Pasiarski, Marcin; Korona-Glowniak, Izabela; Maj, Maciej; Surdacka, Agata; Grafka, Agnieszka; Stelmach-Gołdyś, Agnieszka; Zgurski, Michał; Góźdź, Stanisław; Malm, Anna; Grabarczyk, Piotr; Starosławska, Elżbieta

    2015-01-01

    Epstein-Barr virus (EBV) is a ubiquitous γ-herpesvirus that infects more than 90% of the world population. The potential involvement of EBV in the clinical course of chronic lymphocytic leukemia (CLL) remains unexplained. The aim of this study was to determine whether EBV-DNA load in the peripheral blood mononuclear cells (PBMCs) of CLL patients may influence heterogeneity in the course of the disease. The study included peripheral blood samples from 115 previously untreated patients with CLL (54 women and 61 men) and 40 healthy controls (16 women and 24 men). We analyzed the association between the EBV-DNA load in PBMCs and the stage of the disease, adverse prognostic factors, and clinical outcome. Detectable numbers of EBV-DNA copies in PBMCs were found in 62 out of 115 CLL patients (53.91%). The EBV-DNA copy number/μg DNA was significantly higher in patients who required early implementation of treatment, presented with lymphocyte count doubling time DNA copy number/μg DNA showed significant positive correlation with the concentrations of lactate dehydrogenase (LDH) and beta-2-microglobulin. We have shown that in CLL patients, higher EBV-DNA copy number predicted shorter survival and shorter time to disease progression, and it was associated with other established unfavorable prognostic factors. This suggests that EBV may negatively affect the outcome of CLL.

  16. Ruta 6 selectively induces cell death in brain cancer cells but proliferation in normal peripheral blood lymphocytes: A novel treatment for human brain cancer.

    Science.gov (United States)

    Pathak, Sen; Multani, Asha S; Banerji, Pratip; Banerji, Prasanta

    2003-10-01

    Although conventional chemotherapies are used to treat patients with malignancies, damage to normal cells is problematic. Blood-forming bone marrow cells are the most adversely affected. It is therefore necessary to find alternative agents that can kill cancer cells but have minimal effects on normal cells. We investigated the brain cancer cell-killing activity of a homeopathic medicine, Ruta, isolated from a plant, Ruta graveolens. We treated human brain cancer and HL-60 leukemia cells, normal B-lymphoid cells, and murine melanoma cells in vitro with different concentrations of Ruta in combination with Ca3(PO4)2. Fifteen patients diagnosed with intracranial tumors were treated with Ruta 6 and Ca3(PO4)2. Of these 15 patients, 6 of the 7 glioma patients showed complete regression of tumors. Normal human blood lymphocytes, B-lymphoid cells, and brain cancer cells treated with Ruta in vitro were examined for telomere dynamics, mitotic catastrophe, and apoptosis to understand the possible mechanism of cell-killing, using conventional and molecular cytogenetic techniques. Both in vivo and in vitro results showed induction of survival-signaling pathways in normal lymphocytes and induction of death-signaling pathways in brain cancer cells. Cancer cell death was initiated by telomere erosion and completed through mitotic catastrophe events. We propose that Ruta in combination with Ca3(PO4)2 could be used for effective treatment of brain cancers, particularly glioma.

  17. Divergent effect of cobalt and beryllium salts on the fate of peripheral blood monocytes and T lymphocytes.

    Science.gov (United States)

    Paladini, Fabiana; Cocco, Elisa; Potolicchio, Ilaria; Fazekasova, Henrieta; Lombardi, Giovanna; Fiorillo, Maria Teresa; Sorrentino, Rosa

    2011-02-01

    Occupational exposure to metals such as cobalt and beryllium represents a risk factor for respiratory health and can cause immune-mediated diseases. However, the way they act may be different. We show here that the two metals have a divergent effect on peripheral T lymphocytes and monocytes: BeSO(4) induces cell death in monocytes but not in T lymphocytes, which instead respond by producing Interferon gamma (IFN-γ); conversely, CoCl(2) induces apoptosis in T lymphocytes but not in monocytes. Interestingly, both metals induce p53 overexpression but with a dramatic different outcome. This is because the effect of p53 in CoCl(2)-treated monocytes is counteracted by the antiapoptotic activity of cytoplasmic p21(Cip1/WAF1), the activation of nuclear factor κB, and the inflammasome danger signaling pathway leading to the production of proinflammatory cytokines. However, CoCl(2)-treated monocytes do not fully differentiate into macrophage or dendritic cells, as inferred by the lack of expression of CD16 and CD83, respectively. Furthermore, the expression of HLA-class II molecules, as well as the capability of capturing and presenting the antigens, decreased with time. In conclusion, cobalt keeps monocytes in a partially activated, proinflammatory state that can contribute to some of the pathologies associated with the exposure to this metal.

  18. X射线诱发外周血淋巴细胞TCR基因突变研究%X-ray induced TCR gene mutation of peripheral blood lymphocytes

    Institute of Scientific and Technical Information of China (English)

    侯殿俊; 马娅; 刘伟; 乔建维; 卢峰; 商希梅; 李洁清; 封丽

    2009-01-01

    objective To study the TCR gene mutation in peripheral blood lymphocytes induced by X-ray exposure using cultured lymphocytes cloning method.Methods Freshly isolated peripheral lymphocytes from healthy aduh donors were irradiated with X-ray in doses ranging from 0 to 8 Gy and cultured with interleukin2 and phytohemagglutinin for 7 days.The mutant frequencies of TCR gene(TCR MF)were detected by flow cytonletry and the dose response curves were fitted.Results TCR MF increased with the dose going up.An aquadratic polynomial dose response model was fitted.Conclusions TCR gene mutation could which serve as a potential biological dosimeter.It might be applied for the estimation of biological dose in emergency exposure.%目的 用培养法研究X射线诱发的人外周血淋巴细胞TCR基因突变情况.方法 以不同剂量(0~8 Gy)的X射线照射新鲜分离的健康成人外周血淋巴细胞,植物血凝素、白细胞介素2(IL-2)协同刺激培养7 d,流式细胞术检测TCR基因突变频率(TCR MF),并拟合剂量效应关系.结果 随着照射剂量的增加,TCR基因突变频率随之上升,最佳拟合曲线为二次多项式模型.结论 TCR基因突变可作为辐射生物剂量计,用于急性辐射照射生物剂量的估算.

  19. Genotoxic effects of a particular mixture of acetamiprid and alpha-cypermethrin on chromosome aberration, sister chromatid exchange, and micronucleus formation in human peripheral blood lymphocytes.

    Science.gov (United States)

    Kocaman, Ayşe Yavuz; Topaktaş, Mehmet

    2010-04-01

    The genotoxic effects of a particular mixture of acetamiprid (Acm, neonicotinoid insecticide) and alpha-cypermethrin (alpha-cyp, pyrethroid insecticide) on human peripheral lymphocytes were examined in vitro by chromosomal aberrations (CAs), sister chromatid exchange (SCE), and micronucleus (MN) tests. The human peripheral lymphocytes were treated with 12.5 + 2.5, 15 + 5, 17.5 + 7.5, and 20 + 10 microg/mL of Acm+alpha-cyp, respectively, for 24 and 48 h. The mixture of Acm+alpha-cyp induced the CAs and SCEs at all concentrations and treatment times when compared with both the control and solvent control and these increases were concentration-dependent in both treatment times. MN formation was significantly induced at 12.5 + 2.5, 15 + 5, 17.5 + 7.5, microg/mL of Acm+alpha-cyp when compared with both controls although these increases were not concentration-dependent. Binuclear cells could not be detected sufficiently in the highest concentration of the mixture (20 + 10 microg/mL) for both the 24- and 48-h treatment times. Mitotic index (MI), proliferation index (PI) and nuclear division index (NDI) significantly decreased because of the cytotoxic and cytostatic effects of the mixture, at all concentrations for two treatment periods. Significant decreases in MI and PI were concentration dependent at both treatment times. The decrease in NDI was also concentration-dependent at 48-h treatment period. In general, Acm+alpha-cyp inhibited nuclear division more than positive control, mitomycin C (MMC) and showed a higher cytostatic effect than MMC. Furthermore, in this article, the results of combined effects of Acm+alpha-cyp were compared with the results of single effects of Acm or alpha-cyp (Kocaman and Topaktas,2007,2009, respectively). In conclusion, the particular mixture of Acm+alpha-cyp synergistically induced the genotoxicity/cytotoxicity in human peripheral blood lymphocytes.

  20. Increased micronucleus frequency in peripheral blood lymphocytes contributes to cancer risk in the methyl isocyanate-affected population of Bhopal.

    Science.gov (United States)

    Senthilkumar, Chinnu Sugavanam; Akhter, Sameena; Malla, Tahir Mohiuddin; Sah, Nand Kishore; Ganesh, Narayanan

    2015-01-01

    The Bhopal gas tragedy involving methyl isocyanate (MIC) is one of the most horrific industrial accidents in recent decades. We investigated the genotoxic effects of MIC in long-term survivors and their offspring born after the 1984 occurrence. There are a few cytogenetic reports showing genetic damage in the MIC-exposed survivors, but there is no information about the associated cancer risk. The same is true about offspring. For the first time, we here assessed the micronucleus (MN) frequency using cytokinesis-blocked micronucleus (CBMN) assay to predict cancer risk in the MIC-affected population of Bhopal. A total of 92 healthy volunteers (46 MIC- affected and 46 controls) from Bhopal and various regions of India were studied taking gender and age into consideration. Binucleated lymphocytes with micronuclei (BNMN), total number of micronuclei in lymphocytes (MNL), and nuclear division index (NDI) frequencies and their relationship to age, gender and several lifestyle variabilities (smoking, alcohol consumption and tobacco-chewing) were investigated. Our observations showed relatively higher BNMN and MNL (Pexposure to MIC. Briefly, the observed cytogenetic damage to the MIC-affected could contribute to cancer risk, especially in the EF and FOE.

  1. Diversity of V delta-J delta gene rearrangement in peripheral blood lymphocytes and intrathecal IgG synthesis in multiple sclerosis.

    Science.gov (United States)

    Michałowska-Wender, G; Nowak, J; Losy, J; Januszkiewicz, D; Wender, M

    1999-01-01

    The object of the study is a comparison of intrathecal IgG synthesis and gamma/delta TCR genes rearrangement in multiple sclerosis. The subgroup of 13 cases with intrathecal IgG synthesis and positive oligoclonal bands was compared with 8 cases with IgG index below 0.75 and with undetectable oligoclonal bands. TCR gene rearrangement was studied in peripheral blood lymphocytes by PCR analysis. In majority of cases of the first group the V delta-J delta junctional repertoire was restricted as evidenced by oligoclonal rearrangement. Monoclonal pattern of rearrangement was also established in some cases concerning V delta 1-J delta 1 and V delta 5-J delta 1. In all cases with one exception, demonstrating IgG index < 0.75 and with negative oligoclonal bands in CSF the oligo- or polyclonal pattern of V delta-J delta gene rearrangement was noticed. It is therefore suggested that subset T and B lymphocytes may undergo clonal expansion in MS as evidenced by restricted pattern of V delta-J delta rearrangement and intrathecal oligoclonal IgG synthesis, respectively. Oligoclonal expansion at certain B and T cells may occur due to stimulation by an antigen related to MS pathogen.

  2. Defect in lectin-induced interleukin 2 production by peripheral blood lymphocytes of patients with invasive urinary bladder carcinoma

    DEFF Research Database (Denmark)

    Bubeník, J; Kieler, J; Tromholt, V

    1988-01-01

    The production of interleukin 2 (IL-2) by phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) from 21 patients with transitional cell carcinoma of the urinary bladder (BTCC) and 16 control blood donors was measured with a solid phase enzyme immunoassay based on the dual...

  3. Lower HIV provirus levels are associated with more APOBEC3G protein in blood resting memory CD4+ T lymphocytes of controllers in vivo.

    Directory of Open Access Journals (Sweden)

    Mariapia De Pasquale

    Full Text Available Immunodeficiency does not progress for prolonged periods in some HLA B57- and/or B27-positive subjects with human immunodeficiency virus type 1 (HIV infection, even in the absence of antiretroviral therapy (ART. These "controllers" have fewer HIV provirus-containing peripheral blood mononuclear cells than "non-controller" subjects, but lymphocytes that harbor latent proviruses were not specifically examined in studies to date. Provirus levels in resting memory cells that can serve as latent reservoirs of HIV in blood were compared here between controllers and ART-suppressed non-controllers. APOBEC3G (A3G, a cellular factor that blocks provirus formation at multiple steps if not antagonized by HIV virion infectivity factor (Vif, was also studied. HLA-linked HIV control was associated with less provirus and more A3G protein in resting CD4+ T central memory (Tcm and effector memory (Tem lymphocytes (provirus: p = 0.01 for Tcm and p = 0.02 for Tem; A3G: p = 0.02 for Tcm and p = 0.02 for Tem. Resting memory T cells with the highest A3G protein levels (>0.5 RLU per unit of actin had the lowest levels of provirus (<1,000 copies of DNA per million cells in vivo (p = 0.03, Fisher's exact test. Using two different experimental approaches, Vif-positive viruses with more A3G were found to have decreased virion infectivity ex vivo. These results raise the hypothesis that HIV control is associated with increased cellular A3G that may be packaged into Vif-positive virions to add that mode of inhibition of provirus formation to previously described adaptive immune mechanisms for HIV control.

  4. High Viral Loads of Epstein-Barr Virus DNA in Peripheral Blood of Patients with Chronic Lymphocytic Leukemia Associated with Unfavorable Prognosis.

    Directory of Open Access Journals (Sweden)

    Ewelina Grywalska

    Full Text Available Epstein-Barr virus (EBV is a ubiquitous γ-herpesvirus that infects more than 90% of the world population. The potential involvement of EBV in the clinical course of chronic lymphocytic leukemia (CLL remains unexplained. The aim of this study was to determine whether EBV-DNA load in the peripheral blood mononuclear cells (PBMCs of CLL patients may influence heterogeneity in the course of the disease. The study included peripheral blood samples from 115 previously untreated patients with CLL (54 women and 61 men and 40 healthy controls (16 women and 24 men. We analyzed the association between the EBV-DNA load in PBMCs and the stage of the disease, adverse prognostic factors, and clinical outcome. Detectable numbers of EBV-DNA copies in PBMCs were found in 62 out of 115 CLL patients (53.91%. The EBV-DNA copy number/μg DNA was significantly higher in patients who required early implementation of treatment, presented with lymphocyte count doubling time <12 months, displayed CD38-positive or ZAP-70-positive phenotype, and with the del(11q22.3 cytogenetic abnormality. Furthermore, the EBV-DNA copy number/μg DNA showed significant positive correlation with the concentrations of lactate dehydrogenase (LDH and beta-2-microglobulin. We have shown that in CLL patients, higher EBV-DNA copy number predicted shorter survival and shorter time to disease progression, and it was associated with other established unfavorable prognostic factors. This suggests that EBV may negatively affect the outcome of CLL.

  5. Effect of acitretin in combined with NB-UVB on T lymphocyte subsets in the peripheral blood of patients with psoriasis vulgaris

    Institute of Scientific and Technical Information of China (English)

    Rui-Ju Niu

    2015-01-01

    Objective:To explore the effect of acitretin in combined with NB-UVB on T lymphocyte subsets in the peripheral blood in patients with psoriasis vulgaris.Methods:A total of 90 patients with psoriasis vulgaris who were admitted in our department from May, 2013 to May, 2015 were included in the study and randomized into the observation group and the control group. The patients in the control group were given acitretin after dinner, 10 mg/time, twice each day, for 6 weeks. The patients in the observation group were given acitretin in combined with UVB, each irradiation for 30 min, three times every week, for 6 weeks. The changes of CD3+, CD4+, and CD8+ in the peripheral blood, PASI score, and the clinical efficacy before and after treatment in the two groups were observed.Results:The elevation degree of CD3+ and CD4+ after treatment in the observation group was significantly obvious that in the control group, while the declining degree of CD8+ was also significantly obvious that in the control group. The decreased degree of PASI score after treatment in the observation group was significantly superior to that in the control group. The total effective rate in the observation group (93.33%) was significantly superior to that in the control group (80.00%).Conclusions:Acitretin in combined with UVB in the treatment of psoriasis vulgaris can play its role by altering the imbalance state of T lymphocyte immune function. The combined treatment can enhance the therapeutic effect, therefore, it deserves to be widely recommended in the clinic.

  6. Effect of Metallothionein on Cell Cycle, Apoptosis Rate and Subsets Distribution of Lymphocytes in Peripheral Blood of Dairy Cattle under Heat Stress

    Institute of Scientific and Technical Information of China (English)

    Jiajie LUO; Jun FANG; Lili LI; Bin ZHANG; LiZhuan WU; Zijun LI; Ying PENG; JueXin FAN; XinYi LAN; JinShun ZHAN

    2013-01-01

    [Objective] This study aimed to research the effect of metal othionein on cellcycle, apoptosis rate and subsets distribution of lymphocytes in peripheral blood of dairy cows under heat stress, so as to perfect the regulative mechanism re-searches of MT to anti-heat stress. [Method] Twenty lactating Chinese Holstein cows were randomly divided into four groups (A, B, C and D), and injected with 0, 4.0, 8.0 and 12.0 mg Zn-metal othionein, respectively by intravenous route. Blood sam-ples were col ected at 1st, 16th, 31st, 46th and 61st day, and the dynamic changes of cellcycle, apoptosis rate and subsets distribution of lymphocytes were determined. [Result] The apoptosis rate of cells in group B and C was lower than those in group A by 26.63% (P>0.05) and 24.84% (P>0.05) respectively. The number of cells in the G0/G1 phage in trial groups was increased and the number of cells in the S and G2/M phages tended to decrease, but there were no significant differences (P>0.05). The number of CD3+ T cellin three trial groups was greater than those in group A by 7.02% (P>0.05), 5.45% (P>0.05) and 3.85% (P>0.05) respectively, while the number of CD4+ T cellin trial groups was higher than those in control group by 31.04% (P<0.05), 35.68% (P<0.05) and 39.34% (P<0.05) respectively. The number of CD8+ T celland the levels of CD4+/CD8+ in trial groups were increased observ-ably, but significant difference (P<0.05) was observed in the levels of CD4+/CD8+between groups A and C only. It demonstrated that exogenous Zn-metal othionein can decrease apoptosis rate, improve cellcycle and regulate subsets distribution of lymphocytes in dairy cattle in a dose-dependent manner. [Conclusion] This study wil provide scientific basis for safe utilization of MT in dairy industry.

  7. Effect of lignin supplementation of a diet contaminated with Fusarium mycotoxins on blood and intestinal lymphocyte subpopulations in chickens.

    Science.gov (United States)

    Revajová, Viera; Levkut, Mikuláš; Levkutová, Mária; Bořutová, Radka; Grešaková, Lubomíra; Košiková, Božena; Leng, Lubomír

    2013-09-01

    The objective of the study was to investigate the effects of lignin supplementation of a diet contaminated with the Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEA) on peripheral blood leukocytes and duodenal immunocompetent cells in broiler chickens. From day 1 after hatching, all chickens were fed an identical control diet for two weeks. Then chickens of Group 1 continued to be fed the control diet, whereas Group 2 was fed the same diet supplemented with lignin at 0.5% level. Simultaneously, Group 3 started to receive a diet contaminated with DON (2.95 mg kg-1) and ZEA (1.59 mg kg-1), while Group 4 received an identical contaminated diet supplemented with 0.5% lignin for further two weeks. Samples of blood and duodenal tissue were collected from 6 birds of each group at 4 weeks of age. Neither counts of white blood cells nor phagocytic function in the peripheral blood were significantly affected in the mycotoxin- and/or lignin-treated birds. As compared to the control, increased numbers of IgM-bearing cells were found in the peripheral blood in Group 3 fed the contaminated diet (P contaminated diet supplemented with lignin (P contaminated diet led to reduced numbers of duodenal CD4+ cells, in Group 2 treated only with lignin the number of duodenal CD4+ cells was increased. Lignin enrichment of the contaminated diet did not eliminate the mycotoxin-induced reduction in the number of duodenal CD4+ cells. The results suggest that dietary supplementation of lignin as an indigestible compound to poultry feed may increase the density of some intestinal immunocompetent cells without exerting effects on that in the peripheral blood. However, when added to a diet contaminated with Fusarium mycotoxins, lignin did not prevent the mycotoxin-induced changes in the numbers of blood and intestinal immunocompetent cells.

  8. Increased Fas and Bcl-2 expression on peripheral blood T and B lymphocytes from juvenile-onset systemic lupus erythematosus, but not from juvenile rheumatoid arthritis and juvenile dermatomyositis.

    Science.gov (United States)

    Liphaus, Bernadete L; Kiss, Maria H B; Carrasco, Solange; Goldenstein-Schainberg, Claudia

    2006-01-01

    Defective regulation of apoptosis may play a role in the development of autoimmune diseases. Fas and Bcl-2 proteins are involved in the control of apoptosis. The aims of this study were to determine the expression of Fas antigen and Bcl-2 protein on peripheral blood T and B lymphocytes from patients with juvenile-onset systemic lupus erythematosus (JSLE), juvenile rheumatoid arthritis (JRA) and juvenile dermatomyositis (JDM). Thirty-eight patients with JSLE, 19 patients with JRA, 10 patients with JDM and 25 healthy controls entered the study. Freshly isolated peripheral blood mononuclear cells (PBMC) were stained for lymphocyte markers CD3, CD4, CD8, CD19 and for Fas and Bcl-2 molecules. Expressions were measured by three-color flow cytometry. Statistical analysis was performed using Kruskal-Wallis test. Percentages of freshly isolated T lymphocytes positively stained for Fas protein from JSLE patients were significantly increased compared to healthy controls, patients with JRA and patients with JDM. Percentages of B lymphocytes positive for Fas from JSLE patients were higher than healthy controls and JRA patients. In addition, Fas expression on T cells from patients with JRA was increased compared to JDM patients. Otherwise, Fas expression on T and B cells from JRA and JDM patients were similar to healthy controls. MFI of Bcl-2 positive T lymphocytes from JSLE patients were significantly increased compared to healthy controls and JRA patients. MFI of Bcl-2 protein on B lymphocytes from JSLE patients was similar to healthy controls and patients with JRA and JDM. Bcl-2 expression did not differ between JRA and JDM patients and healthy controls. In conclusion, increased expression of Fas and Bcl-2 proteins observed in circulating T and B lymphocytes from patients with JSLE, but not from patients with JRA and JDM, suggests that abnormalities of apoptosis may be related to the pathogenesis of JSLE and probably are not a result of chronic inflammation.

  9. Increased Fas and Bcl-2 Expression on Peripheral Blood T and B Lymphocytes from Juvenile-Onset Systemic Lupus Erythematosus, but not from Juvenile Rheumatoid Arthritis and Juvenile Dermatomyositis

    Directory of Open Access Journals (Sweden)

    Bernadete L. Liphaus

    2006-01-01

    Full Text Available Defective regulation of apoptosis may play a role in the development of autoimmune diseases. Fas and Bcl-2 proteins are involved in the control of apoptosis. The aims of this study were to determine the expression of Fas antigen and Bcl-2 protein on peripheral blood T and B lymphocytes from patients with juvenile-onset systemic lupus erythematosus (JSLE, juvenile rheumatoid arthritis (JRA and juvenile dermatomyositis (JDM. Thirty-eight patients with JSLE, 19 patients with JRA, 10 patients with JDM and 25 healthy controls entered the study. Freshly isolated peripheral blood mononuclear cells (PBMC were stained for lymphocyte markers CD3, CD4, CD8, CD19 and for Fas and Bcl-2 molecules. Expressions were measured by three-color flow cytometry. Statistical analysis was performed using Kruskal–Wallis test. Percentages of freshly isolated T lymphocytes positively stained for Fas protein from JSLE patients were significantly increased compared to healthy controls, patients with JRA and patients with JDM. Percentages of B lymphocytes positive for Fas from JSLE patients were higher than healthy controls and JRA patients. In addition, Fas expression on T cells from patients with JRA was increased compared to JDM patients. Otherwise, Fas expression on T and B cells from JRA and JDM patients were similar to healthy controls. MFI of Bcl-2 positive T lymphocytes from JSLE patients were significantly increased compared to healthy controls and JRA patients. MFI of Bcl-2 protein on B lymphocytes from JSLE patients was similar to healthy controls and patients with JRA and JDM. Bcl-2 expression did not differ between JRA and JDM patients and healthy controls. In conclusion, increased expression of Fas and Bcl-2 proteins observed in circulating T and B lymphocytes from patients with JSLE, but not from patients with JRA and JDM, suggests that abnormalities of apoptosis may be related to the pathogenesis of JSLE and probably are not a result of chronic inflammation.

  10. Concentration-Dependent Protection by Ethanol Extract of Propolis against γ-Ray-Induced Chromosome Damage in Human Blood Lymphocytes

    Directory of Open Access Journals (Sweden)

    A. Montoro

    2011-01-01

    Full Text Available Radioprotection with natural products may be relevant to the mitigation of ionizing radiation-induced damage in mammalian systems; in this sense, propolis extracts have shown effects such as antioxidant, antitumoral, anti-inflammatory, and immunostimulant. We report for the first time a cytogenetic study to evaluate the radioprotective effect, in vitro, of propolis against radiation-induced chromosomal damage. Lymphocytes were cultured with increasing concentrations of ethanol extract of propolis (EEP, including 20, 40, 120, 250, 500, 750, 1000, and 2000 μg mL−1 and then exposed to 2 Gy γ-rays. A significant and concentration-dependent decrease is observed in the frequency of chromosome aberrations in samples treated with EEP. The protection against the formation of dicentrics was concentration-dependent, with a maximum protection at 120 μg mL−1 of EEP. The observed frequency of dicentrics is described as negative exponential function, indicating that the maximum protectible fraction of dicentrics is approximately 44%. Free radical scavenging and antioxidant activities are the mechanisms that these substances use to protect cells from ionizing radiation.

  11. Peripheral blood lymphocyte telomere length as a predictor of response to immunosuppressive therapy in childhood aplastic anemia.

    Science.gov (United States)

    Sakaguchi, Hirotoshi; Nishio, Nobuhiro; Hama, Asahito; Kawashima, Nozomu; Wang, Xinan; Narita, Atsushi; Doisaki, Sayoko; Xu, Yinyan; Muramatsu, Hideki; Yoshida, Nao; Takahashi, Yoshiyuki; Kudo, Kazuko; Moritake, Hiroshi; Nakamura, Kazuhiro; Kobayashi, Ryoji; Ito, Etsuro; Yabe, Hiromasa; Ohga, Shouichi; Ohara, Akira; Kojima, Seiji

    2014-08-01

    Predicting the response to immunosuppressive therapy could provide useful information to help the clinician define treatment strategies for patients with aplastic anemia. In our current study, we evaluated the relationship between telomere length of lymphocytes at diagnosis and the response to immunosuppressive therapy in 64 children with aplastic anemia, using flow fluorescence in situ hybridization. Median age of patients was ten years (range 1.5-16.2 years). Severity of the disease was classified as very severe in 23, severe in 21, and moderate in 20 patients. All patients were enrolled in multicenter studies using antithymocyte globulin and cyclosporine. The response rate to immunosuppressive therapy at six months was 52% (33 of 64). The probability of 5-year failure-free survival and overall survival were 56% (95% confidence interval (CI): 41-69%) and 97% (95%CI: 87-99%), respectively. Median telomere length in responders was -0.4 standard deviation (SD) (-2.7 to +3.0 SD) and -1.5 SD (-4.0 to +1.6 (SD)) in non-responders (Paplastic anemia.

  12. [Mutagen influence with different mechanisms of action on DNA global methylation in human whole-blood lymphocytes in vitro].

    Science.gov (United States)

    Smirnikhina, S A; Voronina, E S; Strelnikov, V V; Tanas, A S; Lavrov, A V

    2013-07-01

    Data that support the evidence of mutagens known to cause epigenetic abnormalities that could potentially result in genomic instability and the development of cancer rather than to modifications in the human genome at the gene and chromosomal levels only. The level of global methylation in human lymphocytes in vitro caused by exposure to two mutagens with different mechanisms of action, i.e., dioxidine and methyl methanesulphonate (MMS), was demonstrated in the present study. Global methylation was assessed by methyl-sensitive comet assay. An increase in the level of global methylation to 45.64% was revealed during culturing with dioxidine in a concentration of 0.01 mg/mL (p < 0.001), while the addition of dioxidine in a concentration of 0.1 mg/mL resulted in a decreased level of methylation up to 42.31% (p < 0.001). The addition of M MS in concentrations of 0.0025 and 0.01 mg/mL resulted in minor but significant modifications (p < 0.05) of the global methylation level ranged within natural variations in global methylation. Accordingly, the addition ofdioxidine in the concentration of 0.1 mg/mL might cause genomic instability and might be considered a potential carcinogen.

  13. Spontaneous and radiation-induced micronucleus frequencies in low dose radiation exposed worker's peripheral blood lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Kwon, Hee Kyung; Lee, Hye Jin; Park, Mi Young; Park, Hyun Jin; Kim, Tae Hwan [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of); Ji, Young Hoon; Kim, Ki Sup; Lee, Su Jae; Lee, Yun Sil; Cho, Chul Koo; Choi, Soo Yong; Kang, Chang Mo [Kyungpook National Univ., Daegu (Korea, Republic of)

    2005-07-01

    Many studies have been performed to assess the development and application of potentially useful biodosimetry. At present, although chromosome dicentric assay is a sensitive method for dose estimation, it is laborious and requires enough experience for estimation, and without automation its scope for population screening is limited. Therefore, we need an alternative cytogenetic dosimetry to estimate the absorbed dose of victims after low dose exposure such as radiation accidents in hospital workers and workers of radiation related facilities. An alternative and simple cytogenetic technique is the measurement of the micronucleus frequency in cultured human lymphocytes. The reliability of conventional micronucleus (MN) assays is diminished owing to the inclusion of nondividing cells in the estimate, but this problem has been overcome by the development of the cytokinesisblocked (CB) MN assay. The reliable and ease assays of the cytokinesis blocked-approach are obvious advantages in biological monitoring, but there are no developed recognizable and reliable techniques for biological dosimetry of a low dose exposure until recently. Adaptive response is important in determining the biological responses at low doses of radiation and has the potential to impact the shape of the dose-response relationship. We analyzed the frequency of both spontaneous and in vitro {sup 137}Cs {gamma}-rays-induced MNs to estimate the low dose radiation-exposed workers as a screening test.

  14. Construction of a cytogenetic dose-response curve for low-dose range gamma-irradiation in human peripheral blood lymphocytes using three-color FISH.

    Science.gov (United States)

    Suto, Yumiko; Akiyama, Miho; Noda, Takashi; Hirai, Momoki

    2015-12-01

    In order to estimate biological doses after low-dose ionizing radiation exposure, fluorescence in situ hybridization (FISH) using three differentially colored chromosome painting probes was employed to detect exchange-type chromosome aberrations. A reference dose response curve was constructed using blood samples from a female donor whose lymphocytes consistently exhibited a low frequency of cells at the second mitosis under routine culture conditions. Aberration yields were studied for a total of about 155 thousand metaphases obtained from seven dose-points of gamma irradiations (0, 50, 100, 150, 200, 250 and 300mGy). In situ hybridization was performed using commercially available painting probes for chromosomes 1, 2 and 4. With the aid of an automated image-capturing method, exchange-type aberrations involving painted chromosomes were detected with considerable accuracy and speed. The results on the exchange-type aberrations (dicentrics plus translocations) at the seven dose-points showed a good fit to the linear-quadratic model (y=0.0023+0.0015x+0.0819x(2), P=0.83). A blind test proved the reproducibility of the reference dose-response relationship. In the control experiments using blood samples from another donor, the estimated doses calculated on the basis of the present reference curve were proved to be in good agreement with the actual physical doses applied. The present dose-response curve may serve as a means to assess the individual differences in cytogenetical radio-sensitivities.

  15. Over-Expression of Dopamine D2 Receptor and Inwardly Rectifying Potassium Channel Genes in Drug-Naive Schizophrenic Peripheral Blood Lymphocytes as Potential Diagnostic Markers

    Directory of Open Access Journals (Sweden)

    Ágnes Zvara

    2005-01-01

    Full Text Available Schizophrenia is one of the most common neuropsychiatric disorders affecting nearly 1% of the human population. Current diagnosis of schizophrenia is based on complex clinical symptoms. The use of easily detectable peripheral molecular markers could substantially help the diagnosis of psychiatric disorders. Recent studies showed that peripheral blood lymphocytes (PBL express subtypes of D1 and D2 subclasses of dopamine receptors. Recently, dopamine receptor D3 (DRD3 was found to be over-expressed in schizophrenic PBL and proposed to be a diagnostic and follow-up marker for schizophrenia. In this study we screened PBL of 13 drug-naive/drug-free schizophrenic patients to identify additional markers of schizophrenia. One of the benefits of our study is the use of blood samples of non-medicated, drug-naive patients. This excludes the possibility that changes detected in gene expression levels might be attributed to the medication rather than to the disorder itself. Among others, genes for dopamine receptor D2 (DRD2 and the inwardly rectifying potassium channel (Kir2.3 were found to be over-expressed in microarray analysis. Increased mRNA levels were confirmed by quantitative real-time PCR (QRT-PCR using the SybrGreen method and dual labeled TaqMan probes. The use of both molecular markers allows a more rapid and precise prediction of schizophrenia and might help find the optimal medication for schizophrenic patients.

  16. Over-expression of dopamine D2 receptor and inwardly rectifying potassium channel genes in drug-naive schizophrenic peripheral blood lymphocytes as potential diagnostic markers.

    Science.gov (United States)

    Zvara, Agnes; Szekeres, György; Janka, Zoltán; Kelemen, János Z; Cimmer, Csongor; Sántha, Miklós; Puskás, László G

    2005-01-01

    Schizophrenia is one of the most common neuropsychiatric disorders affecting nearly 1% of the human population. Current diagnosis of schizophrenia is based on complex clinical symptoms. The use of easily detectable peripheral molecular markers could substantially help the diagnosis of psychiatric disorders. Recent studies showed that peripheral blood lymphocytes (PBL) express subtypes of D1 and D2 subclasses of dopamine receptors. Recently, dopamine receptor D3 (DRD3) was found to be over-expressed in schizophrenic PBL and proposed to be a diagnostic and follow-up marker for schizophrenia. In this study we screened PBL of 13 drug-naive/drug-free schizophrenic patients to identify additional markers of schizophrenia. One of the benefits of our study is the use of blood samples of non-medicated, drug-naive patients. This excludes the possibility that changes detected in gene expression levels might be attributed to the medication rather than to the disorder itself. Among others, genes for dopamine receptor D2 (DRD2) and the inwardly rectifying potassium channel (Kir2.3) were found to be over-expressed in microarray analysis. Increased mRNA levels were confirmed by quantitative real-time PCR (QRT-PCR) using the SybrGreen method and dual labeled TaqMan probes. The use of both molecular markers allows a more rapid and precise prediction of schizophrenia and might help find the optimal medication for schizophrenic patients.

  17. Review of the correlation between results of cytogenetic dosimetry from blood lymphocytes and EPR dosimetry from tooth enamel for victims of radiation accidents.

    Science.gov (United States)

    Khvostunov, I K; Ivannikov, A I; Skvortsov, V G; Nugis, V Yu; Golub, E V

    2015-03-01

    The goal of this study was to compare dose estimates from electron paramagnetic resonance (EPR) dosimetry with teeth and cytogenetic dosimetry with blood lymphocytes for 30 victims of radiation accidents. The whole-body exposures estimated by tooth enamel EPR dosimetry were ranging from 0.01 to 9.3 Gy. Study group comprised victims exposed to acute and prolonged irradiation at high and low dose rate in different accidents. Blood samples were taken from each of them for cytogenetic analysis. Aberrations were scored and analysed according to International Atomic Energy Agency (IAEA) guidelines for conventional and FISH analysis. Tooth samples were collected in dental clinics after they had been extracted during ordinary practice. EPR dosimetry was performed according to the IAEA protocol. EPR dosimetry showed good correlation with dosimetry based on chromosomal analysis. All estimations of cytogenetic dose below detection limit coincide with EPR dose estimates within the ranges of uncertainty. The differences between cytogenetic and EPR assays may occur in a case of previous unaccounted exposure, non-homogeneous irradiation and due to contribution to absorbed dose from neutron irradiation.

  18. FIV establishes a latent infection in feline peripheral blood CD4+ T lymphocytes in vivo during the asymptomatic phase of infection

    Directory of Open Access Journals (Sweden)

    Murphy Brian

    2012-02-01

    Full Text Available Abstract Background Feline immunodeficiency virus (FIV is a lentivirus of cats that establishes a lifelong persistent infection with immunologic impairment. Results In an approximately 2 year-long experimental infection study, cats infected with a biological isolate of FIV clade C demonstrated undetectable plasma viral loads from 10 months post-infection onward. Viral DNA was detected in CD4+CD25+ and CD4+CD25- T cells isolated from infected cats whereas viral RNA was not detected at multiple time points during the early chronic phase of infection. Viral transcription could be reactivated in latently infected CD4+ T cells ex vivo as demonstrated by detectable FIV gag RNA and 2-long terminal repeat (LTR circle junctions. Viral LTR and gag sequences amplified from peripheral blood mononuclear cells during early and chronic stages of infection demonstrated minimal to no viral sequence variation. Conclusions Collectively, these findings are consistent with FIV latency in peripheral blood CD4+ T cells isolated from chronically infected cats. The ability to isolate latently FIV-infected CD4+ T lymphocytes from FIV-infected cats provides a platform for the study of in vivo mechanisms of lentiviral latency.

  19. Emotional stress induced by parachute jumping enhances blood nerve growth factor levels and the distribution of nerve growth factor receptors in lymphocytes.

    Science.gov (United States)

    Aloe, L; Bracci-Laudiero, L; Alleva, E; Lambiase, A; Micera, A; Tirassa, P

    1994-10-25

    We examined the plasma nerve growth factor (NGF) level and the distribution of NGF receptors in peripheral lymphocytes of young soldiers (mean age, 20-24 yr) experiencing the thrill of a novice about to make their first parachute jumps. Blood was collected from soldiers who knew they were selected to jump (n = 26), as well as from soldiers who knew they were not selected (n = 17, controls). The former group was sampled the evening before the jump and 20 min after landing. Compared with controls, NGF levels increased 84% in prejump and 107% in postjump sampling. Our studies also showed that the increase of NGF levels preceded the increase of plasma cortisol and adrenocorticotropic hormone. No changes in the baseline levels of circulating interleukin 1 beta or tumor necrosis factor were found, suggesting that the increased levels of NGF were not correlated with change in these cytokines. Moreover, immunofluorescence analysis demonstrated that parachuting stress enhances the distribution of low-affinity p75LNGFR and high-affinity p140trkA NGF receptors in circulating peripheral blood mononuclear cells. These observations suggest that the release of NGF might be involved in the activation of cells of the immune system and is most probably associated with homeostatic adaptive mechanisms, as previously shown for stressed rodents.

  20. Distinct immunological mechanisms of CTLA-4 and PD-1 blockade revealed by analyzing TCR usage in blood lymphocytes

    Science.gov (United States)

    Robert, Lidia; Harview, Christina; Emerson, Ryan; Wang, Xiaoyan; Mok, Stephen; Homet, Blanca; Comin-Anduix, Begonya; Koya, Richard C; Robins, Harlan; Tumeh, Paul C; Ribas, Antoni

    2014-01-01

    Targeting immune inhibitory receptors has brought excitement, innovation and hope to cancer patients. Our recent work revealed the immunological effects of blocking the CTLA4 and PD-1 immune checkpoints on T cell receptor usage among peripheral blood cells, and further uncovers how the expansion of the T cell repertoire matches the immunotoxicity profile of the therapy. PMID:25083336

  1. Blood levels of CD11b+ memory T lymphocytes are selectively upregulated in patients with active rheumatoid arthritis

    DEFF Research Database (Denmark)

    Nielsen, H; Petersen, A A; Skjødt, H;

    1999-01-01

    The adhesion molecules CD11b (a beta2-integrin component) and CD54 (ICAM-1) on blood leukocytes were studied by flow cytometry in patients with rheumatoid arthritis (RA). The fractions of CD4+ cells co-expressing CD11b were elevated in 16 patients with active RA compared with those in 16 RA...

  2. The profiles of gamma-H2AX along with ATM/DNA-PKcs activation in the lymphocytes and granulocytes of rat and human blood exposed to gamma rays.

    Science.gov (United States)

    Wang, Jing; Yin, Lina; Zhang, Junxiang; Zhang, Yaping; Zhang, Xuxia; Ding, Defang; Gao, Yun; Li, Qiang; Chen, Honghong

    2016-08-01

    Establishing a rat model suitable for γ-H2AX biodosimeter studies has important implications for dose assessment of internal radionuclide contamination in humans. In this study, γ-H2AX, p-ATM and p-DNA-PKcs foci were enumerated using immunocytofluorescence method, and their protein levels were measured by Western blot in rat blood lymphocytes and granulocytes exposed to γ-rays compared with human blood lymphocytes and granulocytes. It was found that DNA double-strand break repair kinetics and linear dose responses in rat lymphocytes were similar to those observed in the human counterparts. Moreover, radiation induced clear p-ATM and p-DNA-PKcs foci formation and an increase in ratio of co-localization of p-ATM or p-DNA-PKcs with γ-H2AX foci in rat lymphocytes similar to those of human lymphocytes. The level of γ-H2AX protein in irradiated rat and human lymphocytes was significantly reduced by inhibitors of ATM and DNA-PKcs. Surprisingly, unlike human granulocytes, rat granulocytes with DNA-PKcs deficiency displayed a rapid accumulation, but delayed disappearance of γ-H2AX foci with essentially no change from 10 h to 48 h post-irradiation. Furthermore, inhibition of ATM activity in rat granulocytes also decreased radiation-induced γ-H2AX foci formation. In comparison, human granulocytes showed no response to irradiation regarding γ-H2AX, p-ATM or p-DNA-PKcs foci. Importantly, incidence of γ-H2AX foci in lymphocytes after total-body radiation of rats was consistent with that of in vitro irradiation of rat lymphocytes. These findings show that rats are a useful in vivo model for validation of γ-H2AX biodosimetry for dose assessment in humans. ATM and DNA-PKcs participate together in DSB repair in rat lymphocytes similar to that of human lymphocytes. Further, rat granulocytes, which have the characteristic of delayed disappearance of γ-H2AX foci in response to radiation, may be a useful experimental system for biodosimetry studies.

  3. Rosette formation of pig T lymphocytes with sheep erythrocytes.

    Science.gov (United States)

    Escajadillo, C; Binns, R M

    1975-01-01

    The relationship of sheep RBC rosette formation to density of thymus and blood lymphocytes was investigated. Thymocyte density was unimodal and cells of all densities rosetted equally. Blood lymphocyte density was bimodal with most rosette-forming cells in the denser ficoll layers. Papain treatment of SRBC increases rosette formation with blood lymphocytes while apparently maintaining specificity of T cells.

  4. Hydrogen peroxide-induced oxidative damage in peripheral blood lymphocytes from rats chronically treated with corticosterone: The protective effect of oxytocin treatment.

    Science.gov (United States)

    Stanić, Dušanka; Plećaš-Solarović, Bosiljka; Petrović, Jelena; Bogavac-Stanojević, Nataša; Sopić, Miron; Kotur-Stevuljević, Jelena; Ignjatović, Svetlana; Pešić, Vesna

    2016-08-25

    Contemporary lifestyle is commonly associated with chronic stress, an environmental factor contributing to development of various psychological and somatic disorders. Increased levels of glucocorticoids, observed in the chronic stress, induce the production of reactive oxygen species leading to genotoxicity. The aim of this study was to investigate whether chronic administration of oxytocin (OXY) 10 IU/400 μL/day, s.c., for 14 days, a hormone presumed to exert antioxidant effect, may prevent DNA damage in the comet assay of peripheral blood lymphocytes of Wistar rats treated chronically with corticosterone (CORT) 100 mg/L ad libitum, per os, for 21 days, as well as, to influence some plasma oxidative stress parameters, i.e. levels of total lipid hydroperoxide (LOOH), and malondialdehyde (MDA), and the activity of antioxidative enzyme superoxide dismutase (SOD). Even though there was no reduction in overall number of damaged cells after oxytocin treatment only, the marked increase in total comet score (TCS) after incubation with H2O2 in CORT group compared to controls, was absent in the CORT + OXY experimental group. Furthermore, significant decrease of highly damaged cells compared to corticosterone group was noted. Chronic oxytocin administration thus protected lymphocytes from high intensity damage that leads to cellular death. In addition, treatment with OXY along with CORT, significantly decreased concentration of LOOH in plasma, and increased SOD compared to CORT treatment only. This finding corresponds well with current reports on beneficial effects of OXY in conditions of HPA axis hyperactivity, and supports the hypothesis of OXY-mediated antioxidant action.

  5. Dicentric chromosomes and gamma-H2AX foci formation in lymphocytes of human blood samples exposed to a CT scanner: a direct comparison of dose response relationships.

    Science.gov (United States)

    Golfier, Sven; Jost, Gregor; Pietsch, Hubertus; Lengsfeld, Philipp; Eckardt-Schupp, Friederike; Schmid, Ernst; Voth, Matthias

    2009-02-01

    Experiments using the induction of dicentric chromosomes (dicentrics) as well as the gamma-H2AX foci formation in lymphocytes of blood samples from a healthy donor were performed to directly evaluate the radiation sensitivity of both biological endpoints. For computed tomography scans at dose levels from 0.025 to 1 Gy, a linear-quadratic dose-response relationship for dicentrics and a linear dose-response relationship for gamma-H2AX foci were obtained. The coefficients of the dose-response relationship for dicentrics are alpha = (3.76 +/- 0.29) x 10(-2) Gy(-1) and beta = (5.54 +/- 0.45) x 10(-2) Gy(-2), the linear coefficient for gamma-H2AX foci is (7.38 +/- 0.11) Gy(-1). The findings indicate that scoring of dicentrics as well as microscopic analysis of gamma-H2AX foci are sensitive methods to quantify a radiation-induced biological damage at low doses. However, since gamma-H2AX foci can be partially repaired within a few hours, biological damages present for days or even months, which constitute the clinically relevant endpoints, can only be quantified reliably by scoring of chromosome aberrations. Thus currently the quantification of dicentrics or reciprocal translocations remains the recommended method for estimating the effect of exposures to low dose levels of radiation ('biological dosimetry'). However, owing to the high radiation sensitivity of the gamma-H2AX foci assay observed in the present study, further investigations on the effectiveness of low-linear energy transfer radiation qualities in producing gamma-H2AX foci in lymphocytes from healthy donors should be performed.

  6. [Effects of recombinant interleukin-2 on several characteristics of functional activity of lymphocytes from the lymph nodes regional to tumor and mononuclear cells of peripheral blood in cancer patients].

    Science.gov (United States)

    Semenova-Kobzar', R A; Kushko, L Ia; Iakhimovich, L V; Protsyk, V S; Tolstopiatov, B A; Konovalenko, V F; Berezhnaia, N M

    1990-01-01

    The level of endogenous production of IL-2 by lymphocytes of lymph nodes regional to tumour and by mononuclear cells of peripheral blood, proliferative response of these cell to recombinant IL-2, as well as a modifying influence of autologous serum and actively proliferating bioptats of autologous tumours on enumerated parameters have been studied in cancer patients (tumours of the head and neck and locomotor system). Regional IL-2-dependent immunotherapy of malignant tumors with obligatory preliminary testing for individual sensitivity of the tumor bioptat to the influence of the RIL-2 and RIL-2 activated lymphocytes is shown to be promising.

  7. Use of Gene Expression Profiles of Peripheral Blood Lymphocytes to Distinguish BRCA1 Mutation Carriers in High Risk Breast Cancer Families

    Directory of Open Access Journals (Sweden)

    Marie-Laure Vuillaume

    2009-01-01

    Full Text Available Mutations in two major genes, BRCA1 and BRCA2, account for up to 30% of families with hereditary breast cancer. Unfortunately, in most families there is little to indicate which gene should be targeted first for mutation screening, which is labor intensive, time consuming and often prohibitively expensive. As BRCA1 is a tumor suppressor gene involved in various cellular processes, heterozygous mutations could deregulate dependent pathways, such as DNA damage response, and disturb transcriptional activity of genes involved in the downstream signaling cascade. We investigated gene expression profiling in peripheral blood lymphocytes to evaluate this strategy for distinguishing BRCA1 mutation carriers from non-carriers. RNA from whole blood samples of 15 BRCA1 mutation carriers and 15 non-carriers from BRCA1 or BRCA2 families were hybridized to Agilent Technologies Whole Human Genome OligoMicroarrays (4 × 44 K multiplex format containing 41,000 unique human genes and transcripts. Gene expression data were analyzed with Welch’s t-tests and submitted to hierarchical clustering (GeneSpring GX software, Agilent Technologies. Statistical analysis revealed a slight tendency for 133 genes to be differentially expressed between BRCA1 mutation carriers and non-carriers. However, hierarchical clustering of these genes did not accurately discriminate BRCA1 mutation carriers from non-carriers. Expression variation for these genes according to BRCA1 mutation status was weak. In summary, microarray profiling of untreated whole blood does not appear to be informative in identifying breast cancer risk due to BRCA1 mutation.

  8. Effects of Exercise on Behavior and Peripheral Blood Lymphocyte Apoptosis in a Rat Model of Chronic Fatigue Syndrome

    Institute of Scientific and Technical Information of China (English)

    邹军; 苑建齐; 吕爽; 屠嘉衡

    2010-01-01

    This study examined the effects of exercise on behavior and peripheral blood leukocyte apoptosis in a rat model of chronic fatigue syndrome(CFS).Thirty-six healthy male Sprague-Dawley rats were equally randomized into 3 groups:the control group,CFS model group and the exercise group in terms of body weight.A total of 25 rats entered the final statistical analysis due to 11 deaths during the study.CFS model was established by subjecting the rats in CFS model group and exercise group to electric shock,chronic...

  9. [The effect of small doses of ionizing radiation on the physico-chemical characteristics of the membranes of peripheral blood lymphocytes of rats].

    Science.gov (United States)

    Prishchep, S G; Gerasimovich, N V; Bulanova, K Ia; Miliutin, A A

    2000-01-01

    The fluorescence probe method was used for investigating the physical state of a total lipid phase of a bi-layer and an annular (near-protein) zone of the membrane lipids of lymphocytes in a peripheral blood of rats on the 10th day after a whole-body acute and chronic gamma-exposure to doses of 0.25, 0.5, and 1.0 Gy. It was discovered that exposure to doses of 0.25, 0.5, and 1.0 Gy revealed no reliable distinctions in the parameters of a physical state of the lipid component of membranes within the given period of observation if compared with those of controls. However chronic exposure to the same doses caused the increase in hydrophobicity of the total lipid phase of the membrane bi-layer with no change in the polarity of an annular lipid. The near-protein zone of lipids revealed a local decrease in microviscosity while the fluidity of total lipids of a membrane bi-layer remained unaltered. A detected change of tryptophan fluorescence of the membrane proteins after exposing them to small dosed has also been carried out.

  10. Indoor air pollution from biomass burning activates Akt in airway cells and peripheral blood lymphocytes: a study among premenopausal women in rural India.

    Science.gov (United States)

    Mondal, Nandan K; Roy, Amrita; Mukherjee, Bidisha; Das, Debangshu; Ray, Manas R

    2010-12-01

    Biomass burning is a major source of indoor air pollution in rural India. The authors investigated in this study whether cumulative exposures to biomass smoke cause activation of the serine/threonine kinase Akt in airway cells and peripheral blood lymphocytes (PBL). For this, the authors enrolled 87 premenopausal (median age 34 years), nonsmoking women who used to cook with biomass (wood, dung, crop wastes) and 85 age-matched control women who cooked with cleaner fuel liquefied petroleum gas. Immunocytochemical and immunoblotting assays revealed significantly higher levels of phosphorylated forms of Akt protein (p-Akt(ser473) and p-Akt(thr308)) in PBL, airway epithelial cells, alveolar macrophages, and neutrophils in sputum of biomass-using women than control. Akt activation in biomass users was associated with marked rise in generation of reactive oxygen species and concomitant depletion of superoxide dismutase. Measurement of particulate matter having a diameter of less than 10 and 2.5 µm in indoor air by real-time aerosol monitor showed 2 to 4 times more particulate pollution in biomass-using households, and Akt activation was positively associated with particulate pollution after controlling potential confounders. The findings suggest that chronic exposure to biomass smoke activates Akt, possibly via generation of oxidative stress.

  11. In-vitro assessment of cytotoxicity of halloysite nanotubes against HepG2, HCT116 and human peripheral blood lymphocytes.

    Science.gov (United States)

    Ahmed, Farrukh Rafiq; Shoaib, Muhammad Harris; Azhar, Mudassar; Um, Soong Ho; Yousuf, Rabia Ismail; Hashmi, Shahkamal; Dar, Ahsana

    2015-11-01

    Halloysite is a clay mineral with chemical similarity to kaolin, a pharmaceutical ingredient. It consists of mainly aluminosilicate nanotubular particles in the size range of ∼ 200-1000 nm. Many studies have tried to empirically explore this novel clay for its potential in drug delivery systems but no work has yet studied its cytotoxicity from the perspective of oral drug delivery system. In this study, the halloysite nanotubes (HNTs) were subjected to size distribution analyses, which reveal more than 50% of nanotubes in the size range of 500 nm and rest mainly in the sub micrometer range. HNTs were then evaluated for in-vitro cytotoxicity against HCT116 (colorectal carcinoma) and HepG2 (hepatocellular carcinoma) cells which represent the earliest entry point and the first accumulating organ, respectively, for nanoparticles en-route to systemic circulation after oral delivery. Moreover, HNTs were tested for their cytogenetic toxicity against human peripheral blood lymphocytes. Both these results collectively indicated that HNTs are generally safe at practical concentrations of excipients for oral dosage forms.

  12. Maternal exposure to air pollution before and during pregnancy related to changes in newborn's cord blood lymphocyte subpopulations. The EDEN study cohort

    Directory of Open Access Journals (Sweden)

    Baïz Nour

    2011-11-01

    Full Text Available Abstract Background Toxicants can cross the placenta and expose the developing fetus to chemical contamination leading to possible adverse health effects, by potentially inducing alterations in immune competence. Our aim was to investigate the impacts of maternal exposure to air pollution before and during pregnancy on newborn's immune system. Methods Exposure to background particulate matter less than 10 μm in diameter (PM10 and nitrogen dioxide (NO2 was assessed in 370 women three months before and during pregnancy using monitoring stations. Personal exposure to four volatile organic compounds (VOCs was measured in a subsample of 56 non-smoking women with a diffusive air sampler during the second trimester of pregnancy. Cord blood was analyzed at birth by multi-parameter flow cytometry to determine lymphocyte subsets. Results Among other immunophenotypic changes in cord blood, decreases in the CD4+CD25+ T-cell percentage of 0.82% (p = 0.01, 0.71% (p = 0.04, 0.88% (p = 0.02, and 0.59% (p = 0.04 for a 10 μg/m3 increase in PM10 levels three months before and during the first, second and third trimester of pregnancy, respectively, were observed after adjusting for confounders. A similar decrease in CD4+CD25+ T-cell percentage was observed in association with personal exposure to benzene. A similar trend was observed between NO2 exposure and CD4+CD25+ T-cell percentage; however the association was stronger between NO2 exposure and an increased percentage of CD8+ T-cells. Conclusions These data suggest that maternal exposure to air pollution before and during pregnancy may alter the immune competence in offspring thus increasing the child's risk of developing health conditions later in life, including asthma and allergies.

  13. High ACSL5 Transcript Levels Associate with Systemic Lupus Erythematosus and Apoptosis in Jurkat T Lymphocytes and Peripheral Blood Cells

    Science.gov (United States)

    2011-01-01

    Background Systemic lupus erythematosus (SLE) is a prototypical autoimmune disease in which increased apoptosis and decreased apoptotic cells removal has been described as most relevant in the pathogenesis. Long-chain acyl-coenzyme A synthetases (ACSLs) have been involved in the immunological dysfunction of mouse models of lupus-like autoimmunity and apoptosis in different in vitro cell systems. The aim of this work was to assess among the ACSL isoforms the involvement of ACSL2, ACSL4 and ACSL5 in SLE pathogenesis. Findings With this end, we determined the ACSL2, ACSL4 and ACSL5 transcript levels in peripheral blood mononuclear cells (PBMCs) of 45 SLE patients and 49 healthy controls by quantitative real time-PCR (q-PCR). We found that patients with SLE had higher ACSL5 transcript levels than healthy controls [median (range), healthy controls = 16.5 (12.3–18.0) vs. SLE = 26.5 (17.8–41.7), P = 3.9×10 E-5] but no differences were found for ACSL2 and ACSL4. In in vitro experiments, ACSL5 mRNA expression was greatly increased when inducing apoptosis in Jurkat T cells and PBMCs by Phorbol-Myristate-Acetate plus Ionomycin (PMA+Io). On the other hand, short interference RNA (siRNA)-mediated silencing of ACSL5 decreased induced apoptosis in Jurkat T cells up to the control levels as well as decreased mRNA expression of FAS, FASLG and TNF. Conclusions These findings indicate that ACSL5 may play a role in the apoptosis that takes place in SLE. Our results point to ACSL5 as a potential novel functional marker of pathogenesis and a possible therapeutic target in SLE. PMID:22163040

  14. High ACSL5 transcript levels associate with systemic lupus erythematosus and apoptosis in Jurkat T lymphocytes and peripheral blood cells.

    Directory of Open Access Journals (Sweden)

    Antonio Catalá-Rabasa

    Full Text Available BACKGROUND: Systemic lupus erythematosus (SLE is a prototypical autoimmune disease in which increased apoptosis and decreased apoptotic cells removal has been described as most relevant in the pathogenesis. Long-chain acyl-coenzyme A synthetases (ACSLs have been involved in the immunological dysfunction of mouse models of lupus-like autoimmunity and apoptosis in different in vitro cell systems. The aim of this work was to assess among the ACSL isoforms the involvement of ACSL2, ACSL4 and ACSL5 in SLE pathogenesis. FINDINGS: With this end, we determined the ACSL2, ACSL4 and ACSL5 transcript levels in peripheral blood mononuclear cells (PBMCs of 45 SLE patients and 49 healthy controls by quantitative real time-PCR (q-PCR. We found that patients with SLE had higher ACSL5 transcript levels than healthy controls [median (range, healthy controls = 16.5 (12.3-18.0 vs. SLE = 26.5 (17.8-41.7, P = 3.9×10 E-5] but no differences were found for ACSL2 and ACSL4. In in vitro experiments, ACSL5 mRNA expression was greatly increased when inducing apoptosis in Jurkat T cells and PBMCs by Phorbol-Myristate-Acetate plus Ionomycin (PMA+Io. On the other hand, short interference RNA (siRNA-mediated silencing of ACSL5 decreased induced apoptosis in Jurkat T cells up to the control levels as well as decreased mRNA expression of FAS, FASLG and TNF. CONCLUSIONS: These findings indicate that ACSL5 may play a role in the apoptosis that takes place in SLE. Our results point to ACSL5 as a potential novel functional marker of pathogenesis and a possible therapeutic target in SLE.

  15. Peripheral blood lymphocyte cell subsets in subjects with chronic obstructive pulmonary disease: association with smoking, IgE and lung function

    NARCIS (Netherlands)

    J.W. de Jong (Jan Willem); B. van der Belt-Gritter; G.H. Koëter (Gerard); D.S. Postma (Dirkje)

    1997-01-01

    textabstractIn contrast to the numerous studies which show that lymphocytes play an important role in the pathogenesis of asthma, few studies have investigated the role of lymphocytes in the pathogenesis of chronic obstructive pulmonary disease (COPD). The aim of the present study

  16. DNA repair in modeled microgravity: Double strand break rejoining activity in human lymphocytes irradiated with {gamma}-rays

    Energy Technology Data Exchange (ETDEWEB)

    Mognato, Maddalena, E-mail: maddalena.mognato@unipd.it [Dipartimento di Biologia, Universita di Padova, via U. Bassi 58 B, 35121 Padova (Italy); Girardi, Cristina; Fabris, Sonia [Dipartimento di Biologia, Universita di Padova, via U. Bassi 58 B, 35121 Padova (Italy); Celotti, Lucia [Dipartimento di Biologia, Universita di Padova, via U. Bassi 58 B, 35121 Padova (Italy); Laboratori Nazionali di Legnaro, INFN, Padova (Italy)

    2009-04-26

    Cell response to ionising radiation depends, besides on genetic and physiological features of the biological systems, on environmental conditions occurring during DNA repair. Many data showed that microgravity, experienced by astronauts during space flights or modeled on Earth, causes apoptosis, cytoskeletal alteration, cell growth inhibition, increased frequency of mutations and chromosome aberrations. In this study, we analysed the progression of the rejoining of double strand breaks (DSBs) in human peripheral blood lymphocytes (PBLs) irradiated with {gamma}-rays and incubated in static condition (1g) or in modeled microgravity (MMG). {gamma}-H2AX foci formation and disappearance, monitored during the repair incubation, showed that the kinetics of DSBs rejoining was different in the two gravity conditions. The fraction of foci-positive cells decreased slower in MMG than in 1g at 6 and 24 h after irradiation (P < 0.01) and the mean number of {gamma}-H2AX foci per nucleus was significantly higher in MMG than in 1g at the same time-points (P < 0.001). In the same samples we determined apoptotic level and the rate of DSB rejoining during post-irradiation incubation. A significant induction of apoptosis was observed in MMG at 24 h after irradiation (P < 0.001), whereas at shorter times the level of apoptosis was slightly higher in MMG respect to 1g. In accordance with the kinetics of {gamma}-H2AX foci, the slower rejoining of radiation-induced DSBs in MMG was observed by DNA fragmentation analyses during the repair incubation; the data of pulsed-field gel electrophoresis assay showed that the fraction of DNA released in the gel was significantly higher in PBL incubated in MMG after irradiation with respect to cells maintained in 1g. Our results provide evidences that MMG incubation during DNA repair delayed the rate of radiation-induced DSB rejoining, and increased, as a consequence, the genotoxic effects of ionising radiation.

  17. Whole-exome sequencing of DNA from peripheral blood mononuclear cells (PBMC and EBV-transformed lymphocytes from the same donor

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    Delgrosso Kathleen

    2011-09-01

    Full Text Available Abstract Background The creation of lymphoblastoid cell lines (LCLs through Epstein-Barr virus (EBV transformation of B-lymphocytes can result in a valuable biomaterial for cell biology research and a renewable source of DNA. While LCLs have been used extensively in cellular and genetic studies, the process of cell transformation and expansion during culturing may introduce genomic changes that may impact their use and the interpretation of subsequent genetic findings. Results We performed whole exome sequencing on a tetrad family using DNA derived from peripheral blood mononuclear cells (PBMCs and LCLs from each individual. We generated over 4.7 GB of mappable sequence to a 125X read coverage per sample. An average of 19,354 genetic variants were identified. Comparison of the two DNA sources from each individual showed an average concordance rate of 95.69%. By lowering the variant calling parameters, the concordance rate between the paired samples increased to 99.82%. Sanger sequencing of a subset of the remaining discordant variants did confirm the presence of de novo mutations arising in LCLs. Conclusions By varying software stringency parameters, we identified 99% concordance between DNA sequences derived from the two different sources from the same donors. These results suggest that LCLs are an appropriate representation of the genetic material of the donor and suggest that EBV transformation can result in low-level generation of de novo mutations. Therefore, use of PBMC or early passage EBV-transformed cells is recommended. These findings have broad-reaching implications, as there are thousands of LCLs in public biorepositories and individual laboratories.

  18. Inhibitory effect of heparin-derived oligosaccharides on secretion of interleukin-4 and interleukin-5 from human peripheral blood T lymphocytes

    Institute of Scientific and Technical Information of China (English)

    Sheng-Li Ji; Hui-Fei Cui; Feng Shi; Yan-Qing Chi; Ji-Chao Cao; Mei-Yu Geng; Hua-Shi Guan

    2004-01-01

    AIM: To investigate the inhibitory effect of heparin-derived oligosaccharides (Oligs) on secretion of interleukin-4 (IL-4)and interleukin-5 (IL-5) from human peripheral blood T lymphocytes (PBTLs).METHODS: Oligs were prepared by three different heparin depolymerization methods and separated by gel filtration chromatography. PBTLs from ten adult patients with allergic eosinophilic gastroenteritis were treated with phytahematoagglutinin (PHA) and Oligs. The supernatants from the cell culture of PBTLs were harvested and subjected to the deterrnination of IL-4 and IL-5 contents by ELISA method.RESULTS: At the concentration of 5 μg/mL, Oligs with different Mr had different effects on the secretion of IL-4 and IL-5. The tetrasaccharide with Mr of 1 142, produced by depolymerizing heparin with hydrogen peroxide, had the strongest inhibitory effect on the secretion of IL-4. It decreased the IL-4 content from 375.6±39.2 ng/L (PHA group) to 12.5±5.7 ng/L (P<0.01). The hexasaccharide with Mr of 1 806, produced by depolymerizing heparin with β-elimination method, had the strongest inhibitory effect on the secretion of IL-5. It decreased the IL-5 content from 289.2±33.4 ng/L (PHA group) to 22.0±5.2 ng/L (P<0.01).CONCLUSION: The inhibitory activity of Oligs on the secretion of IL-4 and IL-5 from human PBTLs closely depends on their molecular structure, and there may be an essential structure to act as an inhibitor. The most effective inhibitors of IL-4 and IL-5 secretion are tetrasaccharides and hexasaccharides, respectively.

  19. Dose response of multiple parameters for calyculin A-induced premature chromosome condensation in human peripheral blood lymphocytes exposed to high doses of cobalt-60 gamma-rays.

    Science.gov (United States)

    Lu, Xue; Zhao, Hua; Feng, Jiang-Bin; Zhao, Xiao-Tao; Chen, De-Qing; Liu, Qing-Jie

    2016-09-01

    Many studies have investigated exposure biomarkers for high dose radiation. However, no systematic study on which biomarkers can be used in dose estimation through premature chromosome condensation (PCC) analysis has been conducted. The present study aims to screen the high-dose radiation exposure indicator in calyculin A-induced PCC. The dose response of multiple biological endpoints, including G2/A-PCC (G2/M and M/A-PCC) index, PCC ring (PCC-R), ratio of the longest/shortest length (L/L ratio), and length and width ratio of the longest chromosome (L/B ratio), were investigated in calyculin A-induced G2/A-PCC spreads in human peripheral blood lymphocytes exposed to 0-20Gy (dose-rate of 1Gy/min) cobalt-60 gamma-rays. The G2/A-PCC index was decreased with enhanced absorbed doses of 4-20Gy gamma-rays. The G2/A PCC-R at 0-12Gy gamma-rays conformed to Poisson distribution. Three types of PCC-R were scored according to their shape and their solidity or hollowness. The frequencies of hollow PCC-R and PCC-R including or excluding solid ring in G2/A-PCC spreads were enhanced with increased doses. The length and width of the longest chromosome, as well as the length of the shortest chromosome in each G2/M-PCC or M/A-PCC spread, were measured. All L/L or L/B ratios in G2/M-PCC or M/A-PCC spread increased with enhanced doses. A blind test with two new irradiated doses was conducted to validate which biomarker could be used in dose estimation. Results showed that hollow PCC-R and PCC-R including solid ring can be utilized for accurate dose estimation, and that hollow PCC-R was optimal for practical application.

  20. T lymphocyte subsets of the umbilical cord blood of dogs Subpopulações de linfócitos t no sangue de cordão umbilical de cães

    Directory of Open Access Journals (Sweden)

    M.L.B. Cápua

    2009-08-01

    Full Text Available The hematological parameters red blood cells (RBC and total white blood cells (WBC counts, hematocrit, hemoglobin concentration, and RBC indexes (median corpuscular volume and median corpuscular hemoglobin concentration were determined and T CD5+ lymphocytes and CD4+ and CD8+ subpopulations of the umbilical cord blood (UCB of dogs were quantified by the cytofluorimetric technique. Nine adult Beagles, from two do five-year old, were used as control. The umbilical cord blood (UCB was collected from 20 neonate dogs. The method for the UCB collection was adequate to obtain sufficient quantity of blood for the accomplishment of the hematological analyses and lymphocyte quantification. Cytoscopic preparations of the UCB suggested high erythropoietic activity. There was no difference for the global leukocyte and lymphocyte counts between the groups. UCB T lymphocyte counts were lower than those obtained for adult dogs. The proportion of CD4:CD8 showed a great dominance of T CD4+ cells over T CD8+ lymphocytes in UCB.Determinaram-se os valores hematológicos da contagem de hemácias, contagem total de leucócitos, hematócrito, concentração de hemoglobina e os índices hematimétricos (volume corpuscular médio e concentração de hemoglobina corpuscular média e quantificaram-se os linfócitos T CD5+ e as subpopulações CD4+ e CD8+ do sangue do cordão umbilical (SCU de cães por meio da técnica de citometria de fluxo. Nove cães adultos, da raça Beagle, foram utilizados como controle. O SCU foi colhido de 20 cães neonatos, a termo. O método de colheita de SCU utilizado proporcionou quantidade suficiente de sangue para realização das análises hematológicas e quantificação de linfócitos. As preparações citoscópicas do SCU sugeriram elevada atividade eritropoética. Não houve diferença nas contagens globais de leucócitos e linfócitos entre os grupos. A contagem de linfócitos T no SCU foi mais baixa que a obtida em animais adultos. A

  1. Funcionalidad de la glicoproteína P linfocitaria en la colitis ulcerosa P-glycoprotein functional activity in peripheral blood lymphocytes in ulcerative colitis

    Directory of Open Access Journals (Sweden)

    Catalina M. Cortada

    2009-10-01

    Full Text Available La glicoproteína gp-P, codificada por el MDR1 es una bomba de eflujo transmembrana capaz de movilizar una gran cantidad de fármacos de uso frecuente. Se expresa en la cara luminal del epitelio intestinal, en linfocitos y otros tejidos con función de barrera. MDR1 ha sido postulado como gen candidato en la colitis ulcerosa. El objetivo del presente trabajo fue evaluar la función de la gp-P en linfocitos aislados de sangre periférica de pacientes con colitis en actividad (n = 27 clasificados clínicamente como refractarios (n=16 o respondedores (n = 11 al tratamiento. Se estudió el eflujo de rodamina 123, sustrato de la glicoproteína P, en ausencia y presencia del inhibidor verapamilo (100 μM determinando la fluorescencia intracelular por citometría de flujo. Los resultados se expresaron evaluando el comportamiento de dos marcadores que corresponden al % de células que contienen máxima (M1/mínima (M2 concentración del colorante, reflejando inactividad/actividad de la bomba. Utilizando el test de Kruskal-Wallis y post test de Dunn, se observaron diferencias significativas entre refractarios versus respondedores (p P-glycoprotein (P-gp, encoded by MDR-1, is a transmembrane efflux pump that has been involved in relevant clinical drug transport. It is expressed in lymphocytes, luminal epithelium of colon and other tissues with barrier function. MDR1 was proposed as a candidate gene for ulcerative colitis. The aim of the present work was to investigate the role of P-gp in therapeutic response of ulcerative colitis by studying its functionality in lymphocytes isolated from peripheral blood. Samples were taken from 27 patients with active colitis classified clinically in refractory (n = 16 and responders (n = 11 to treatment. Rhodamine 123 (a fluorescent P-glycoprotein substrate efflux was studied by flow cytometry as absence and presence of an inhibitor (verapamil, 100 μM. Data were expressed evaluating the behaviour of two markers defined

  2. Lymphocyte subset analyses in blood, spleen and lymph nodes of female Sprague-Dawley rats after short or prolonged exposure to a 50 Hz 100-microT magnetic field.

    Science.gov (United States)

    Thun-Battersby, S; Westermann, J; Löscher, W

    1999-10-01

    Based primarily on the results of in vitro studies, it has been suggested that power-line (50 or 60 Hz) magnetic fields (MFs) may reduce immune function, which could lower resistance to infection or cancer. This study was conducted to evaluate the influence of acute and chronic in vivo exposure to a linearly polarized 50 Hz MF on immune function in female Sprague-Dawley rats. Groups of rats were exposed continuously to the MF at a flux density of 100 microT for periods of 3 days, 14 days or 13 weeks. For each exposure period, one control group of rats was sham-exposed together with each MF-exposed group. Experimental end points included analyses of T-lymphocyte subsets as well as other immune cells involved in cell-mediated immune responses, i.e. natural killer (NK) cells, B lymphocytes, macrophages, and granulocytes in blood, spleen and mesenteric lymph nodes. In addition, immunohistochemical methods were used to detect proliferating and apoptotic cells in the various compartments of spleen tissue. The results obtained failed to demonstrate a significant effect of short or prolonged MF exposure on different types of leukocytes, including lymphocyte subsets. Furthermore, the experiments on the in vivo proliferation activity of lymphocytes and the extent of apoptosis in spleen samples did not indicate a difference between the MF-exposed and sham-exposed groups, indicating that MF exposure does not affect the mechanisms involved in the control of lymphocyte homeostasis. The lack of MF effects in the immune tests used in the present in vivo study makes it highly unlikely that MF exposure induces immunotoxicity, at least under the experimental conditions used. However, the data do not exclude the possibility that functional alterations in T-cell responses to mitogens and in NK cell activity as recently described for MF-exposed rodents may be one mechanism involved in the carcinogenic effects of MF exposure observed in some models of co-carcinogenesis.

  3. LYMPHOCYTE APOPTOSIS IN PSORIASIS

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    О. M. Kapuler

    2006-01-01

    Full Text Available Abstract. Forty-two patients with progressive vulgar psoriasis (PASI = 19.7 ± 1.5 and 40 healthy volunteers were under investigation. Psoriatic patients were characterized by increased number of CD4+ CD95+ peripheral blood T lymphocytes, which correlates with clinical psoriatic score, and by increased levels of soluble Fas (sFas in serum, as compared to controls (resp., 1868.1 ± 186.8 pg/ml vs. 1281.4 ± 142.5 pg/ml, PLSD = 0.019. The levels of spontaneous lymphocyte apoptosis and anti-Fas (Mab-induced apoptosis in psoriatic patients did not differ from the controls. However, apoptosis induced by “oxidative stress” (50 M Н202, 4 hrs was depressed in the patients. Moreover, a simultaneous assessment of cell cycle structure (metachromatic staining with Acridine Orange, apoptosis and Fas receptor expression (AnnV-FITC/antiFas mAbs-PE staining following a short-term mitogenic stimulation (PHA-P, 5 µg/ml, 24 hrs were performed. We found no marked differences in mitogenic reactivity, activation-induced apoptosis, and activation-induced Fas receptor expression when studying lymphocytes from healthy donors and psoriatic patients. However, PHA-activated lymphocytes from psoriatic patients displayed a significantly decreased ratio of AnnV+CD95+ to the total AnnV+ subpopulation, thus suggesting a decreased role of Fas-dependent mechanisms of apoptosis during the cell activation. The data obtained confirm a view, that an abnormal lymphocyte “apoptotic reactivity”, which plays a crucial role in the mechanisms of autoimmunity, may also of importance in the pathogenesis of psoriasis.

  4. Transcriptome analysis of the human T lymphocyte cell line Jurkat and human peripheral blood mononuclear cells exposed to deoxynivalenol (DON): New mechanistic insights

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    Katika, Madhumohan R. [RIKILT-Institute of Food Safety, Wageningen University and Research Centre, Wageningen (Netherlands); Department of Health Risk Analysis and Toxicology, Maastricht University (Netherlands); Netherlands Toxicogenomics Centre (Netherlands); Hendriksen, Peter J.M. [RIKILT-Institute of Food Safety, Wageningen University and Research Centre, Wageningen (Netherlands); Netherlands Toxicogenomics Centre (Netherlands); Shao, Jia [RIKILT-Institute of Food Safety, Wageningen University and Research Centre, Wageningen (Netherlands); Department of Health Risk Analysis and Toxicology, Maastricht University (Netherlands); Netherlands Toxicogenomics Centre (Netherlands); Loveren, Henk van [Department of Health Risk Analysis and Toxicology, Maastricht University (Netherlands); National Institute for Public Health and the Environment (RIVM), Bilthoven (Netherlands); Netherlands Toxicogenomics Centre (Netherlands); Peijnenburg, Ad, E-mail: ad.peijnenburg@wur.nl [RIKILT-Institute of Food Safety, Wageningen University and Research Centre, Wageningen (Netherlands); Netherlands Toxicogenomics Centre (Netherlands)

    2012-10-01

    Deoxynivalenol (DON) or vomitoxin is a commonly encountered type-B trichothecene mycotoxin, produced by Fusarium species predominantly found in cereals and grains. DON is known to exert toxic effects on the gastrointestinal, reproductive and neuroendocrine systems, and particularly on the immune system. Depending on dose and exposure time, it can either stimulate or suppress immune function. The main objective of this study was to obtain a deeper insight into DON-induced effects on lymphoid cells. For this, we exposed the human T-lymphocyte cell line Jurkat and human peripheral blood mononuclear cells (PBMCs) to various concentrations of DON for various times and examined gene expression changes by DNA microarray analysis. Jurkat cells were exposed to 0.25 and 0.5 μM DON for 3, 6 and 24 h. Biological interpretation of the microarray data indicated that DON affects various processes in these cells: It upregulates genes involved in ribosome structure and function, RNA/protein synthesis and processing, endoplasmic reticulum (ER) stress, calcium-mediated signaling, mitochondrial function, oxidative stress, the NFAT and NF-κB/TNF-α pathways, T cell activation and apoptosis. The effects of DON on the expression of genes involved in ER stress, NFAT activation and apoptosis were confirmed by qRT-PCR. Other biochemical experiments confirmed that DON activates calcium-dependent proteins such as calcineurin and M-calpain that are known to be involved in T cell activation and apoptosis. Induction of T cell activation was also confirmed by demonstrating that DON activates NFATC1 and induces its translocation from the cytoplasm to the nucleus. For the gene expression profiling of PBMCs, cells were exposed to 2 and 4 μM DON for 6 and 24 h. Comparison of the Jurkat microarray data with those obtained with PBMCs showed that most of the processes affected by DON in the Jurkat cell line were also affected in the PBMCs. -- Highlights: ► The human T cell line Jurkat and human

  5. Cytochrome P450 1A1 and 1B1 in human blood lymphocytes are not suitable as biomarkers of exposure to dioxin-like compounds: polymorphisms and interindividual variation in expression and inducibility.

    Science.gov (United States)

    van Duursen, Majorie B M; Sanderson, J Thomas; van den Berg, Martin

    2005-05-01

    Cytochrome P450 1A1 (CYP1A1) and 1B1 (CYP1B1) are phase I enzymes, the expression of which can be affected by many environmental compounds, including dioxins and dioxin-like compounds. Because CYP1A1 and CYP1B1 expression can easily be determined in peripheral blood lymphocytes, it is often suggested as biomarker of exposure to these compounds. In this study we investigated the interindividual differences in constitutive and induced CYP1A1-catalyzed ethoxyresorufin-O-deethylase (EROD) activity and CYP1A1 and CYP1B1 gene expression in human blood lymphocytes in a group of ten non-smoking females. Freshly isolated lymphocytes were cultured in medium containing the mitogen PHA and were exposed to the most potent dioxin, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or the less potent dioxin-like polychlorinated biphenyl 126 (PCB126). In addition, we determined the occurrence of the CYP1A1 MspI and CYP1B1 Leu432Val polymorphisms. All individuals showed a concentration-dependent increase of EROD activity by TCDD, which was significantly correlated with an increase in CYP1A1, but not CYP1B1 expression. The maximum induced EROD activity by TCDD was very different among the individuals, but the EC50 values were about the same. PCB126 also caused a concentration-dependent increase of EROD activity, but was a factor 100-1000 less potent than TCDD among the individuals. The allele frequencies for CYP1A1 MspI and CYP1B1 Leu432Val reflected a normal Caucasian population and in this study the polymorphisms had no apparent effect on the expression and activity of these enzymes. Our study shows a large interindividual variability in constitutive and induced EROD activity, and CYP1A1 and CYP1B1 expression in human lymphocytes. In addition, dioxin concentrations at which effects were observed in our in vitro study are about 10-fold higher than the human blood levels found in vivo, indicating that EROD activity and CYP1A1 and CYP1B1 expression in human lymphocytes might not be

  6. Low-Dose Total Body Irradiation and Donor Peripheral Blood Stem Cell Transplant Followed by Donor Lymphocyte Infusion in Treating Patients With Non-Hodgkin Lymphoma, Chronic Lymphocytic Leukemia, or Multiple Myeloma

    Science.gov (United States)

    2015-10-30

    Adult Nasal Type Extranodal NK/T-cell Lymphoma; Anaplastic Large Cell Lymphoma; Angioimmunoblastic T-cell Lymphoma; Cutaneous B-cell Non-Hodgkin Lymphoma; Extranodal Marginal Zone B-cell Lymphoma of Mucosa-associated Lymphoid Tissue; Hepatosplenic T-cell Lymphoma; Intraocular Lymphoma; Nodal Marginal Zone B-cell Lymphoma; Noncutaneous Extranodal Lymphoma; Peripheral T-cell Lymphoma; Recurrent Adult Acute Lymphoblastic Leukemia; Recurrent Adult Burkitt Lymphoma; Recurrent Adult Diffuse Large Cell Lymphoma; Recurrent Adult Diffuse Mixed Cell Lymphoma; Recurrent Adult Diffuse Small Cleaved Cell Lymphoma; Recurrent Adult Grade III Lymphomatoid Granulomatosis; Recurrent Adult Immunoblastic Large Cell Lymphoma; Recurrent Adult Lymphoblastic Lymphoma; Recurrent Adult T-cell Leukemia/Lymphoma; Recurrent Cutaneous T-cell Non-Hodgkin Lymphoma; Recurrent Grade 1 Follicular Lymphoma; Recurrent Grade 2 Follicular Lymphoma; Recurrent Grade 3 Follicular Lymphoma; Recurrent Mantle Cell Lymphoma; Recurrent Marginal Zone Lymphoma; Recurrent Mycosis Fungoides/Sezary Syndrome; Recurrent Small Lymphocytic Lymphoma; Refractory Chronic Lymphocytic Leukemia; Refractory Hairy Cell Leukemia; Refractory Multiple Myeloma; Small Intestine Lymphoma; Splenic Marginal Zone Lymphoma; Stage II Multiple Myeloma; Stage III Multiple Myeloma; Testicular Lymphoma; Waldenström Macroglobulinemia

  7. Phytochemical screening of the dichloromethane–ethanolic extract of Eriosema campestre var. macrophylum roots and its antiproliferative effect on human peripheral blood lymphocytes

    Directory of Open Access Journals (Sweden)

    Michaelle G. Santos

    Full Text Available ABSTRACT Eriosema campestre var. macrophylum (Grear Fortunato, Fabaceae, is a native plant of the Brazilian Cerrado and the decoction of its roots has been used by folk medicine for the therapy of inflammatory diseases. In this study we aimed to investigate the effect of the dichloromethane–ethanolic extract of E. campestre roots on the proliferative response of lymphocytes and to examine the profile of IL-2 production. The effect of dichloromethane–ethanolic extract of E. campestre on the proliferation of phytohemagglutinin-stimulated lymphocytes was evaluated by using flow cytometry and the cell supernatants were assayed for IL-2 concentrations by using an enzyme-linked immunosorbent assay. The phytochemical screening of E. campestre roots was performed to determine the main secondary metabolites through chromogenic and precipitation reactions and by using HPLC-PAD. In addition to the presence of subclasses of flavonoids (flavones and flavonols in dichloromethane–ethanolic extract of E. campestre, we observed that the extract induced a concentration-dependent decrease in IL-2 levels on the supernatant of the cell cultures as well as an antiproliferative effect on T lymphocytes, including CD4+ and CD8+ cells. The anti-inflammatory effects attributed to E. campestre by folk medicine may partly be explained by its antiproliferative action on T lymphocytes.

  8. Helper T Lymphocyte Response in the Peripheral Blood of Patients with Intraepithelial Neoplasia Submitted to Immunotherapy with Pegylated Interferon-α

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    Márcia Antoniazi Michelin

    2015-03-01

    Full Text Available Immunotherapy in cancer patients is a very promising treatment and the development of new protocols and the study of the mechanisms of regression is imperative. The objective of this study was to evaluate the production of cytokines in helper T (CD4+ lymphocytes during immunotherapy with pegylated IFN-α in patients with cervical intraepithelial neoplasia (CIN. We conducted a prospective study with 17 patients with CIN II-III using immunotherapy with pegylated IFN-α subcutaneouly weekly, and using flow cytometry we evaluated the peripheric CD4+ T lymphocytes. The results show that in the regression group the patients presented a significant increase in the amount of IFN-γ during the entire immunotherapy, compared with the group without a response. The amount of CD4+ T lymphocytes positive for IL-2, IL-4, IL-10 and TGF-β is significantly lower in patients with good clinical response. The results also demonstrate that patients with regression have a higher amount of intracellular TNF-α in CD4+ T lymphocytes before the start of treatment. Analyzing these data sets, it can be concluded that immunotherapy is a viable clinical treatment for patients with high-grade CIN and that the regression is dependent on the change in the immune response to a Th1 pattern.

  9. Polyhalogenated dibenzo-p-dioxins and dibenzofurans and the immune system. In vitro effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on lymphocytes of venous blood from man and a non-human primate (Callithrix jacchus)

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    Neubert, R.; Helge, H. (Freie Univ. Berlin (Germany, F.R.). Kinderklinik und Poliklinik); Jacob-Mueller, U.; Stahlmann, R.; Neubert, D. (Freie Univ. Berlin (Germany, F.R.). Inst. fuer Toxikologie und Embryonalpharmakologie)

    1991-04-01

    The effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on poke weed mitogen-stimulated proliferation and differentiation of peripheral lymphocytes was studied in vitro with cells from a non-human primate (marmoset monkey, Callithrix jacchus) and from man. Monoclonal antibodies and flow cytometry (FACScan) were used for analysis. The extent of the overall mitogen-stimulated proliferation of isolated lymphocytes in vitro from marmoset blood was only slightly reduced in the presence of TCDD compared to the solvent control (0.01% DMSO). However, incubation with TCDD in the culture medium together with the mitogen led to a pronounced decrease in the percentage of the lymphocyte subset with the surface marker CD4, and a concomitant increase in the percentage of CD8{sup +} cells. The lowest concentration found to be effective in vitro was 1x10{sup -13} M TCDD (25 fg TCDD/ml). When culturing lymphocytes from human blood of different donors under indentical conditions in the presence of TCDD and the mitogen, corresponding effects were observed to those seen with marmoset cells. A closer analysis of the T lymphocyte subsets affected revealed the CD4{sup +}CDw29{sup +} (helper-inducer cells) to be the main target for the action of TCDD. A clear-cut change in the percentage of this subpopulation was induced at concentrations as low as 1x10{sup -13} M TCDD. The development of the IL-2-marker in culture was only slightly affected by TCDD, and concentrations of 1x10{sup -12} M were required to slightly reduce the number of CD2{sup +}CD25{sup +} cells. Special B cells, namely CD20{sup +} (i.e. B{sub 1}) cells, were found to be especially susceptible to the action of TCDD, and clear-cut effects were seen in several experimental series at 1x10{sup -14} M TCDD. The formation of B cells with IgG lambda or kappa chains was also depressed in culture in the presence of TCDD. (orig./MG).

  10. Blood

    Science.gov (United States)

    ... Also, blood is either Rh-positive or Rh-negative. So if you have type A blood, it's either A positive or A negative. Which type you are is important if you need a blood transfusion. And your Rh factor could be important ...

  11. 工频电磁场作业人员周围血淋巴细胞微核分析%Analysis on micronucleus of peripheral blood lymphocytes in workers exposed to power frequency electromagnetic fields

    Institute of Scientific and Technical Information of China (English)

    李丽; 陈慧峰; 李华亮; 黎丽春; 曾广诚; 陈建雄; 温薇; 闫雪华

    2016-01-01

    Objective To examine the effects of long-term exposure to power frequency electromagnetic fields on the occurrence of micronucleus in peripheral blood lymphocytes and the lymphocyte transformation in transformer substation workers .Methods By simple random sampling method , 54 workers exposed to power frequency electromagnetic fields for more than 1.0 year and other 54 non-exposure workers in 500 kV transformer substations were chosen as the exposure group and control group, respectively.The peripheral venous blood of subjects in these two groups was collected , and then the lymphocytes were separated .The micronucleus cell rate , the micronucleus rate and the lymphocyte transformation rate were detected and analyzed .Results Compared with the control group , the micronucleated cell rate , the micronucleus rate , and the lymphocyte transformation rate of the exposure group and its different length of service subgroups (≤10.0 and >10.0 years) showed no statistical significance respectively ( P>0.05).Conclusion The long-term exposure to power frequency electromagnetic fields has no obvious effects on the micronucleus and lymphocyte transformation in the peripheral blood lymphocytes of exposed workers .%目的:探讨长期接触工频电磁场对变电站作业人员周围血淋巴细胞的微核以及淋巴细胞转化的影响。方法采用单纯随机抽样方法,选择54名工龄1.0年以上的500 kV变电站工频电磁场作业人员为接触组,选择54名同单位无职业性工频电磁场接触的人员为对照组,采集2组人群周围静脉血,提取淋巴细胞,检测微核细胞率、微核率以及淋巴细胞转化率,分析其变化情况。结果与对照组比较,接触组及其工龄≤10.0年亚组和工龄>10.0年亚组人群周围血淋巴细胞的微核细胞率、微核率及淋巴细胞转化率均无出现有统计学意义改变( P>0.05)。结论长期接触工频电磁场对作业人员周围血淋巴细胞

  12. DNA damage focus analysis in blood samples of minipigs reveals acute partial body irradiation.

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    Andreas Lamkowski

    Full Text Available Radiation accidents frequently involve acute high dose partial body irradiation leading to victims with radiation sickness and cutaneous radiation syndrome that implements radiation-induced cell death. Cells that are not lethally hit seek to repair ionizing radiation (IR induced damage, albeit at the expense of an increased risk of mutation and tumor formation due to misrepair of IR-induced DNA double strand breaks (DSBs. The response to DNA damage includes phosphorylation of histone H2AX in the vicinity of DSBs, creating foci in the nucleus whose enumeration can serve as a radiation biodosimeter. Here, we investigated γH2AX and DNA repair foci in peripheral blood lymphocytes of Göttingen minipigs that experienced acute partial body irradiation (PBI with 49 Gy (± 6% Co-60 γ-rays of the upper lumbar region. Blood samples taken 4, 24 and 168 hours post PBI were subjected to γ-H2AX, 53BP1 and MRE11 focus enumeration. Peripheral blood lymphocytes (PBL of 49 Gy partial body irradiated minipigs were found to display 1-8 DNA damage foci/cell. These PBL values significantly deceed the high foci numbers observed in keratinocyte nuclei of the directly γ-irradiated minipig skin regions, indicating a limited resident time of PBL in the exposed tissue volume. Nonetheless, PBL samples obtained 4 h post IR in average contained 2.2% of cells displaying a pan-γH2AX signal, suggesting that these received a higher IR dose. Moreover, dispersion analysis indicated partial body irradiation for all 13 minipigs at 4 h post IR. While dose reconstruction using γH2AX DNA repair foci in lymphocytes after in vivo PBI represents a challenge, the DNA damage focus assay may serve as a rapid, first line indicator of radiation exposure. The occurrence of PBLs with pan-γH2AX staining and of cells with relatively high foci numbers that skew a Poisson distribution may be taken as indicator of acute high dose partial body irradiation, particularly when samples are available

  13. Autologous hematopoietic stem cell transplantation in lymphoma patients is associated with a decrease in the double strand break repair capacity of peripheral blood lymphocytes

    Science.gov (United States)

    Lacoste, Sandrine; Bhatia, Smita; Chen, Yanjun; Bhatia, Ravi; O’Connor, Timothy R.

    2017-01-01

    Patients who undergo autologous hematopoietic stem cell transplantation (aHCT) for treatment of a relapsed or refractory lymphoma are at risk of developing therapy related- myelodysplasia/acute myeloid leukemia (t-MDS/AML). Part of the risk likely resides in inherent interindividual differences in their DNA repair capacity (DRC), which is thought to influence the effect chemotherapeutic treatments have on the patient’s stem cells prior to aHCT. Measuring DRC involves identifying small differences in repair proficiency among individuals. Initially, we investigated the cell model in healthy individuals (primary lymphocytes and/or lymphoblastoid cell lines) that would be appropriate to measure genetically determined DRC using host-cell reactivation assays. We present evidence that interindividual differences in DRC double-strand break repair (by non-homologous end-joining [NHEJ] or single-strand annealing [SSA]) are better preserved in non-induced primary lymphocytes. In contrast, lymphocytes induced to proliferate are required to assay base excision (BER) or nucleotide excision repair (NER). We established that both NHEJ and SSA DRCs in lymphocytes of healthy individuals were inversely correlated with the age of the donor, indicating that DSB repair in lymphocytes is likely not a constant feature but rather something that decreases with age (~0.37% NHEJ DRC/year). To investigate the predictive value of pre-aHCT DRC on outcome in patients, we then applied the optimized assays to the analysis of primary lymphocytes from lymphoma patients and found that individuals who later developed t-MDS/AML (cases) were indistinguishable in their DRC from controls who never developed t-MDS/AML. However, when DRC was investigated shortly after aHCT in the same individuals (21.6 months later on average), aHCT patients (both cases and controls) showed a significant decrease in DSB repair measurements. The average decrease of 6.9% in NHEJ DRC observed among aHCT patients was much

  14. Autologous hematopoietic stem cell transplantation in lymphoma patients is associated with a decrease in the double strand break repair capacity of peripheral blood lymphocytes.

    Science.gov (United States)

    Lacoste, Sandrine; Bhatia, Smita; Chen, Yanjun; Bhatia, Ravi; O'Connor, Timothy R

    2017-01-01

    Patients who undergo autologous hematopoietic stem cell transplantation (aHCT) for treatment of a relapsed or refractory lymphoma are at risk of developing therapy related- myelodysplasia/acute myeloid leukemia (t-MDS/AML). Part of the risk likely resides in inherent interindividual differences in their DNA repair capacity (DRC), which is thought to influence the effect chemotherapeutic treatments have on the patient's stem cells prior to aHCT. Measuring DRC involves identifying small differences in repair proficiency among individuals. Initially, we investigated the cell model in healthy individuals (primary lymphocytes and/or lymphoblastoid cell lines) that would be appropriate to measure genetically determined DRC using host-cell reactivation assays. We present evidence that interindividual differences in DRC double-strand break repair (by non-homologous end-joining [NHEJ] or single-strand annealing [SSA]) are better preserved in non-induced primary lymphocytes. In contrast, lymphocytes induced to proliferate are required to assay base excision (BER) or nucleotide excision repair (NER). We established that both NHEJ and SSA DRCs in lymphocytes of healthy individuals were inversely correlated with the age of the donor, indicating that DSB repair in lymphocytes is likely not a constant feature but rather something that decreases with age (~0.37% NHEJ DRC/year). To investigate the predictive value of pre-aHCT DRC on outcome in patients, we then applied the optimized assays to the analysis of primary lymphocytes from lymphoma patients and found that individuals who later developed t-MDS/AML (cases) were indistinguishable in their DRC from controls who never developed t-MDS/AML. However, when DRC was investigated shortly after aHCT in the same individuals (21.6 months later on average), aHCT patients (both cases and controls) showed a significant decrease in DSB repair measurements. The average decrease of 6.9% in NHEJ DRC observed among aHCT patients was much higher

  15. Quantification of micronuclei in blood lymphocytes of patients exposed to gamma radiation for dose absorbed assessment; Quantificacao de micronucleos em linfocitos de pacientes expostas a radiacao gama para a avaliacao da dose absorvida

    Energy Technology Data Exchange (ETDEWEB)

    Barbosa, Isvania Maria Serafim da Silva

    2003-02-15

    Dose assessment in an important step to evaluate biological effects as a result of individual exposure to ionizing radiation. The use of cytogenetic dosimetry based on the quantification of micronuclei in lymphocytes is very important to complement physical dosimetry, since the measurement of absorbed dose cannot be always performed. In this research, the quantification of micronuclei was carried out in order to evaluate absorbed dose as a result of radiotherapy with {sup 60}Co, using peripheral blood samples from 5 patients with cervical uterine cancer. For this purpose, an aliquot of whole blood from the individual patients was added in culture medium RPMI 1640 supplemented with fetal calf serum and phytohaemagglutinin. The culture was incubated for 44 hours. Henceforth, cytochalasin B was added to block the dividing lymphocytes in cytokinesis. The culture was returned to the incubator for further of 28 hours. Thus, cells were harvested, processed and analyzed. Values obtained considering micronuclei frequency after pelvis irradiation with absorption of 0,08 Gy and 1,8 Gy were, respectively, 0,0021 and 0,052. These results are in agreement with some recent researches that provided some standard values related to micronuclei frequency induced by gamma radiation exposure in different exposed areas for the human body. The results presented in this report emphasizes biological dosimetry as an important tool for dose assessment of either total or partial-body exposure to ionizing radiation, mainly in retrospective dose investigation. (author)

  16. Lack of direct immunosuppressive effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on human peripheral blood lymphocyte subsets in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Lang, D.S. (Center for Environmental Medicine and Lung Biology, Univ. of North Carolina, Chapel Hill, NC (United States)); Becker, S. (TCR-Environmental Corp., Inc., Chapel Hill, NC (United States)); Clark, G.C. (National Inst of Environmental Health Science, Lab. of Biochemical Risk Analysis, RTP, NC (United States)); Devlin, R.B. (US Environmental Protection Agency, Health Effects Research Lab., RTP, NC (United States)); Koren, H.S. (US Environmental Protection Agency, Health Effects Research Lab., RTP, NC (United States))

    1994-05-01

    The direct effects of dioxin on human PBL subpopulations have been studied, in order to determine their usefulness as sensitive biomarkers for human dioxin exposure. Lymphocyte cultures from healthy individuals were treated with 10[sup -7] M-10[sup -14] M TCDD in the absence and presence of stimulation with pokeweed mitogen (PWM) or anti-CD3 monoclonal antibody (moAb; OKT3) for 3 days. Cytochrome P450 (CYP1A1) enzyme induction, one of the best studied direct biological effects of TCDD on numerous cell types, was assayed in parallel by ethoxyresorufin-O-deethylase (EROD) activity. Percentages of the different lymphocytes subsets, including CD2 (T cells); CD4; CD45 RA (subpressor-inducer/virgin T cells); CD4 CD29; CD8; CD19 (B cells) as well as interleukin 2 (IL-2) receptor (CD25) and class II antigen (HLA-DR) expression, were analyzed by flow cytometry. DNA synthesis was determined by [sup 3]H-thymidine uptake after 3 days of culture. In the present study, all stimulated lymphocyte cultures showed a dose-dependent significant increase of CYP1A1 activity at dioxin concentrations of 10[sup -7] and 10[sup -9] M. No enzyme activity could be detected at lower concentrations of TCDD. On the other hand, neither alteration in surface marker distribution nor suppression of lymphocyte proliferation could be demonstrated in mitogen-activated cells following any concentration of TCDD treatment. These data suggest that the inducibility of CYP1A1 enzyme activity is not correlated with direct immunotoxic effects in vitro in human PBL. (orig./MG)

  17. Cell death induced by tamoxifen in human blood lymphocytes cultivated in vitro = Morte celular induzida pelo tamoxifeno em linfócitos humanos cultivados in vitro

    Directory of Open Access Journals (Sweden)

    Selma Candelária Genari

    2010-10-01

    Full Text Available Many chemotherapeutic agents with a potential against solid tumors or leukemia can cause lymphopenia. Tamoxifen (TAM is a synthetic non-steroidal anti-estrogen drug employed in female breast cancer treatment. The present study investigated the capacity of TAM to induce cell death in human lymphocytes cultivated in vitro. Lymphocytes were obtained from young (25-30 years; n = 3 and elderly women (58-77 years; n = 3 and cultivated for 24 or 48h, with or without TAM (20 ƒÊM. After the culture, cell viability, immunocytochemical response and ultrastructure were evaluated. TAM affected lymphocytes in a time- dependent manner, and cells obtained from elderly women were the most sensitive to TAM. Immunocytochemicalanalysis evidenced higher frequency of apoptosis in treated cells, and the ultrastructural study revealed autophagic vacuoles, differing from the controls. In summary, the treated lymphocytes were affected by TAM, leading to cell death by apoptosis and autophagy.Muitos agentes quimioterapicos com potencial contra tumores solidos ou leucemias podem causar linfopenia. O Tamoxifeno (TAM e um agente antiestrogeno nao-esteroidal empregado no tratamento de cancer de mama feminino. O presente trabalho investigou a capacidade do TAM em induzir morte celular em linfocitos humanos cultivados in vitro. Oslinfocitos foram obtidos de mulheres jovens (25-30 anos; n = 3 e idosas (58-77 anos; n = 3 e cultivados por 24 ou 48h, com ou sem TAM (20 ƒÊM. Apos a cultura, foram analisadas a viabilidade celular, a resposta imunocitoquimica e a ultraestrutura. Os resultados indicam que o Tamoxifeno induziu morte celular em linfocitos de ambos os grupos, entretanto, as celulas das mulheres idosas apresentaram-se mais sensiveis ao tratamento. A analise imunocitoquimica mostrou maior frequencia de apoptose nas celulas tratadas e o estudo ultraestrutural revelou vacuolos autofagicos nos linfocitos expostos ao Tamoxifeno. Em conclusao, nosso estudo revelou que o TAM

  18. Characterization of the atypical lymphocytes in African swine fever

    Science.gov (United States)

    Karalyan, Z. A.; Ter-Pogossyan, Z. R.; Abroyan, L. O.; Hakobyan, L. H.; Avetisyan, A. S.; Karalyan, N. Yu; Karalova, E. M.

    2016-01-01

    Aim: Atypical lymphocytes usually described as lymphocytes with altered shape, increased DNA amount, and larger size. For analysis of cause of genesis and source of atypical lymphocytes during African swine fever virus (ASFV) infection, bone marrow, peripheral blood, and in vitro model were investigated. Materials and Methods: Atypical lymphocytes under the influence of ASFV were studied for morphologic, cytophotometric, and membrane surface marker characteristics and were used in vivo and in vitro models. Results: This study indicated the increased size, high metabolic activity, and the presence of additional DNA amount in atypical lymphocytes caused by ASFV infection. Furthermore, in atypical lymphocytes, nuclear-cytoplasmic ratio usually decreased, compared to normal lymphocytes. In morphology, they looking like lymphocytes transformed into blasts by exposure to mitogens or antigens in vitro. They vary in morphologic detail, but most of them are CD2 positive. Conclusions: Our data suggest that atypical lymphocytes may represent an unusual and specific cellular response to ASFV infection. PMID:27536044

  19. Characterization of the atypical lymphocytes in African swine fever

    Directory of Open Access Journals (Sweden)

    Z. A. Karalyan

    2016-07-01

    Full Text Available Aim: Atypical lymphocytes usually described as lymphocytes with altered shape, increased DNA amount, and larger size. For analysis of cause of genesis and source of atypical lymphocytes during African swine fever virus (ASFV infection, bone marrow, peripheral blood, and in vitro model were investigated. Materials and Methods: Atypical lymphocytes under the influence of ASFV were studied for morphologic, cytophotometric, and membrane surface marker characteristics and were used in vivo and in vitro models. Results: This study indicated the increased size, high metabolic activity, and the presence of additional DNA amount in atypical lymphocytes caused by ASFV infection. Furthermore, in atypical lymphocytes, nuclear-cytoplasmic ratio usually decreased, compared to normal lymphocytes. In morphology, they looking like lymphocytes transformed into blasts by exposure to mitogens or antigens in vitro. They vary in morphologic detail, but most of them are CD2 positive. Conclusions: Our data suggest that atypical lymphocytes may represent an unusual and specific cellular response to ASFV infection.

  20. Effect of HBe-antigen on Human Peripheral Blood Lymphocytes%乙型肝炎病毒e抗原对人外周血淋巴细胞的影响

    Institute of Scientific and Technical Information of China (English)

    蒋永芳; 唐伟; 马静; 贺波; 李耐萍; 龚国忠

    2012-01-01

    [目的]探索乙型肝炎病毒(HBV)感染后,乙型肝炎e抗原(HBeAg)在免疫调节中的作用和机制.[方法]分离正常人血淋巴细胞,分别加入不同浓度的HBeAg,培养72 h后,计数细胞,流式细胞仪分析T淋巴细胞亚群的改变,同时分析细胞Toll受体(TLR)中TLR3、TLR4和细胞程序性死亡受体-1(PD- 1)的表达率,ELISA分析γ干扰素(IFN-γ)浓度.[结果]淋巴细胞的增殖随HBeAg浓度增加而明显抑制,CD4+和CD8+细胞百分比也相应降低,TLR3、TLR4和PD-1的表达率明显增高,人γ干扰素(IFN-γ)浓度降低.[结论]HBeAg体外能抑制淋巴细胞生长,使T淋巴细胞比例降低,促进PD-1的表达并抑制其活性.%[Objective]To explore the role of HBeAg in the immunological regulation of HBeAg after hepatitis B virus(HBV) infection and its mechanism. [Methods] HBeAg with different concentration were added into lymphocytes isolated from blood of normal people. After cultivation for 72 h, cell count and flow cytome-try were used for analyzing the change of T-lymphocyte subsets. Meanwhile the expression of Toll-like recep-tors(TLR) such as TLR3 and TLR4 and programmed cell death receptor-l(PD-l) were analyzed. ELISA was used to analyze IFN-r concentration. [Results]With the increasing of HBeAg concentration, the proliferation of lymphocytes was inhibited, as well as the percentage of CD4+ and CD8+ lymphocytes correspondingly decreased, but the expression of TLR3 , TLR4 and PD-1 were increased while the expression of INF-r decreased. [Conclusion]HBeAg can inhibit the proliferation of lymphocytes, decrease the percentage of T cells, promote the expression of PD-1 and inhibit its activity in vitro.

  1. Smudge cells in peripheral blood smears did not differentiate chronic lymphocytic leukemia from other B-cell chronic lymphoprolipherative diseases Sombras nucleares no esfregaço do sangue periférico não diferenciam a leucemia linfocítica crônica das outras doenças linfoproliferativas B crônicas

    OpenAIRE

    Matos,Daniel M.; Guilherme Perini; Carlos Kruzich; Rego, Eduardo M.; Falcão, Roberto P.

    2009-01-01

    Smudge cells has been classically associated with chronic lymphocytic leukemia (CLL), but they are found in peripheral blood tests for other chronic B-cell lymphoproliferative diseases (CLD). We investigated whether the percentage of smudge cells in peripheral blood smears can be used in the clinical practice to differentiate CLL from other B-cell CLD. The peripheral blood smears of 63 patients with the diagnosis of CLL and 62 with other B-cell CLD were analyzed. Three hundred cells (both lym...

  2. 间质干细胞对SLE患者外周血T细胞的免疫抑制作用%Immunosuppressive effects of mesenchymal stem cells on peripheral blood T lymphocytes from patients with systemic lupus erythematosus

    Institute of Scientific and Technical Information of China (English)

    殷玉俊; 李晶; 裘影影; 汤郁; 尤海燕; 费晓明; 许文荣

    2009-01-01

    Objective To investigate the immunoregulatory effects of mesenchymal stem cells (MSCs)on peripheral blood T lymphocytes from patients with systemic lupus erythematosus(SLE)in vitro and their potential mechanism.Methods MSCs were isolated from the bone marrow of 3 healthy human volunteers,cultivated and identified.Under phytohemagglutinin(PHA)stimulating,peripheral blood T lymphocytes from 8 patients with SLE were treated with MSCs with the T lymphocyte/MSC ratio being 50:1 in group B and 5:1 in group C or without MSCs(group A).MTT assay was used to detect the proliferation of T lymphocytes.flow cytometry to analyze the expressions of surface markers CD152 and CD28 on T lymphocytes.and real time PCR to measure the mRNA expressions of interleukin-6 and interferon-γ,in the T lymphocytes.Results MSCs could markedly inhibit the proliferation of T lymphocytcs.The proliferation of T lymphocytes expressed as absorbance value at 570 nm was 0.484±0.032 in group B.0.308±0.025 in group C,significantly lower than that in group A(0.765±0.036,both P<0.05),and significant difference was also observed between group C and B(P<0.05).In the case of the percentage of CD28 positive T lymphocytes.group B and C were significantly lower than group A(60.39%±3.92%and 45.05%±3.46%vs 74.73%±3.74%,both P<0.05),and group B significantly differed from group C(P<0.05).MSCs had no obvious effect on the expression of CD152 on T lymphocytes,but significantly suppressed the mRNA expression of interleukin-6 and interferon-γ(both P<0.05).and the suppressive effect was enhanced with the incrgase in MSC count.ConclusionsMSCs exert an immunosuppressive effect on T lymphocytes from patients with SLE,likely through inhibiting the proliferation,CD28 expression,interleukin-6 and interferon-γ mRNA expression of T lymphocytes.%目的 探讨体外骨髓间质干细胞(MSC)对SLE患者外周血T淋巴细胞(T细胞)的免疫调节作用及可能机制.方法 分离培养健康人骨髓MSC并鉴

  3. 两种不同刺激剂对鸡淋巴细胞分泌细胞因子的影响%Effect of Two Kinds of Stimuli on Cytokines Secreted by Chicken Peripheral Blood Lymphocytes

    Institute of Scientific and Technical Information of China (English)

    刘洪明; 王官晓; 王俊卿; 杨玉华; 郭善娜; 徐晓静

    2016-01-01

    [目的]比较不同刺激剂刀豆素A(Con A)和植物凝素(PHA)对鸡淋巴细胞分泌细胞因子的影响。[方法]首先,应用淋巴细胞分离液分离健康鸡的外周血淋巴细胞,应用2种不同的刺激剂Con A和PHA刺激48 h后收集细胞,抽提刺激培养细胞RNA,反转录。其次,应用实时定量PCR方法检测不同刺激剂对鸡淋巴细胞因子mRNA的表达。最后,用分子生物学软件对数据进行比对分析。[结果]分离到纯度和活细胞较高的鸡淋巴细胞。荧光定量PCR结果显示:Con A刺激的淋巴细胞因子表达量比PHA刺激细胞的表达量明显升高(P0.05)。[结论] Con A刺激淋巴细胞后,可以提高细胞因子的表达,刺激效果优于PHA。这些实验结果为进一步体外研究鸡淋巴细胞及其功能提供了方法。%Objective]To compare the effect of two kinds of stimuli,concanavalin A(Con A)and phytohemaggluti-nin(PHA)on cytokine secreted by chicken lymphocyte. [Methods] First,peripheral blood lymphocytes from chick-ens were separated by lymphocyte separation liquid and cultured with Con A and PHA for 48 h,respectively,then the cell RNAs were extracted and the reverse transcription was carry out. Secondly,a real time PCR was performed for detection of the RNA transcription level in the chicken lymphocytes stimulated by Con A and PHA. Finally,the effect of two kinds of stimuli on cytokines secretion was compared with molecular biology software. [Results]Chicken lym-phocytes with higher purity and a survival rate were obtained with the used method. The transcription levels of tested cy-tokines stimulated by Con A were evidently higher than that by PHA(P 0.05)except of IL-4(P < 0.01). [Conclusion]Con A can enhance the transcription levels of cytokine genes and is a be better stimulus for chicken lymphocytes than PHA. All these results provide methods for the further study of chic ken lymphocytes in vitro.

  4. Effects of Stellera chamaejasme L. on the health of human lymphocyte micronucleus rate of peripheral blood%瑞香狼毒对健康人体外外周血淋巴细胞微核率的影响

    Institute of Scientific and Technical Information of China (English)

    张三润; 周好乐; 郑明霞; 布仁其其格; 王金鋒; 张东泽; 郝兴霞

    2014-01-01

    目的:初步探讨瑞香狼毒对健康年轻人体外外周血淋巴细胞内染色体畸变的影响效应。方法在外周血培养过程中加入不同剂量的瑞香狼毒水煎液,显微镜下检测微核率的变化,通过x2检验作统计学分析。结果瑞香狼毒终浓度为2.5、5、10mg/mL实验组的微核出现率分别是1.63‰、5.13‰和7.25‰,对照组微核率为1.50‰,第1个低剂量实验组微核率与对照组比较无统计学差异(P>0.05)。后两个中剂量和高剂量实验组微核率显著高于对照组,并具有统计学差异(P<0.01)。结论超过一定剂量的瑞香狼毒水煎液可能对健康人体外外周血淋巴细胞遗传物质有不同程度的毒性作用。%Objective To discuss toxicity effect of Stellera chamaejasme L.on lymphocyte chromosome in healthy young human peripheral blood.MethodsStellera chamaejasme L.water decoction with different doses was added to the peripheral blood lymphocyte culture fluid, then micronucleus rates were detected under the microscope. The significance tests were conducted byx2 test method. Results The micronucleus rates of the experimental groups in stellera chamaejasme L. exposure concentration of 2.5, 5, 10mg/ml were 1.63‰, 5.13‰ and 7.25‰. The micronucleus rate of control group was 1.50‰. The micronucleus rate of the first low dose experimental group compared with the control group no significant difference(P>0.05). The micronucleus rates of middle and high doses of the experimental groups were significantly higher than that of control group (P<0.01). Conclusion Stellera chamaejasme L. of above a certain dose may exert different degree of damage to genetic material on the health of human peripheral blood lymphocytes.

  5. THE CHANGES OF PERIPHERAL BLOOD T LYMPHOCYTE SUBSETS IN PATIENTS WITH PRIMARY HEPATIC CARCINOMA BOTH PRE-TACE AND POST-TACE

    Institute of Scientific and Technical Information of China (English)

    南克俊; 李春丽; 魏永长; 隋晨光; 刘亚民; 陈葳

    2002-01-01

    Objective To observe the variations of the cellular immunological function in patients with primary hepatic carcinoma both pre-TACE (transcatheter arterial chemoembolization, TACE). Methods T lymphocyte subset CD4, CD8 and CD4/CD7 ratio in 45 patients with primary hepatic carcinoma both pre-TACE and post-TACE were measured by flow Cytometer, and compared with the result of T lymphocyte subset in 19 healthy people as normal control samples. Results The CD4 and CD4/CD8 ratio in patients with primary hepatic carcinoma were significantly lower than those in normal control (P<0.05), while CD8 higher (P<0.05); The CD4 and CD4/CD8 ratio in patients with primary hepatic carcinoma were much more lower after TACE than those before TACE (P<0.05), while CD8 was higher but had no significant difference (P>0.05). Conclusion The cellular immunological function in patients with primary hepatic carcinoma decreased and is much more lower after TACE.

  6. The protective effect of the flavonoids on food-mutagen-induced DNA damage in peripheral blood lymphocytes from colon cancer patients.

    Science.gov (United States)

    Kurzawa-Zegota, Malgorzata; Najafzadeh, Mojgan; Baumgartner, Adolf; Anderson, Diana

    2012-02-01

    The food mutagens IQ (2-amino-3-methylimidazo[4,5-f]quinoline) and PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine) are heterocyclic amines (HCA), generated when heating proteinaceous food. This study investigates the protective potential of the flavonoids quercetin (Q) and rutin (R) against oxidative stress induced in vitro by IQ and PhIP in lymphocytes from healthy individuals and untreated, newly diagnosed colon cancer patients using the Comet assay. In the presence of up to 500μM Q and R, the DNA damage resulting from a high dose of PhIP (75μM) or IQ (150μM) was significantly reduced (P50years of age group and male gender, confounding factors such as smoking, drinking and/or dietary habits were not found to be significant. In conclusion, flavonoids reduced oxidative stress caused by food mutagens in vitro in lymphocytes of healthy individuals and colon cancer patients. Thus, dietary supplementation with flavonoid-rich vegetables and fruits may prove very effective in protecting against oxidative stress.

  7. Subclass of individual IgA-secreting human lymphocytes. Investigation of in vivo pneumococcal polysaccharide-induced and in vitro mitogen-induced blood B cells by monolayer plaque-forming cell assays

    DEFF Research Database (Denmark)

    Heilmann, C; Barington, T; Sigsgaard, T

    1988-01-01

    The subclass of individual human IgA B cells was investigated by means of monolayer plaque-forming cell assays permitting analysis of all IgA-secreting cells as well as of cells secreting IgA anti-pneumococcal polysaccharide antibody. Center cells were examined by indirect immunofluorescence...... that these Ag have an unusually high ability to activate IgA2 B cells, or that the B cells stimulated originate from lymphatic tissues with a high frequency of IgA2 committed cells....... staining with mouse mAb against either of the two IgA subclasses as primary antibodies and FITC-conjugated rabbit anti-mouse Ig as the second antibody. Blood lymphocytes spontaneously secreting IgA (mean 399/10(6) mononuclear cells) produced mainly IgA1 (73%). A similar distribution of subclasses...

  8. A large cohort study reveals the association of elevated peripheral blood lymphocyte-to-monocyte ratio with favorable prognosis in nasopharyngeal carcinoma.

    Directory of Open Access Journals (Sweden)

    Jing Li

    Full Text Available BACKGROUND: Nasopharyngeal carcinoma (NPC is an endemic neoplasm in southern China. Although NPC sufferers are sensitive to radiotherapy, 20-30% of patients finally progress with recurrence and metastases. Elevated lymphocyte-to-monocyte ratio (LMR has been reported to be associated with favorable prognosis in some hematology malignancies, but has not been studied in NPC. The aim of this study was to evaluate whether LMR could predict the prognosis of NPC patients. METHODS: A retrospective cohort of 1,547 non-metastatic NPC patients was recruited between January 2005 and June 2008. The counts for peripheral lymphocyte and monocyte were retrieved, and the LMR was calculated. Receiver operating characteristic curve analysis, univariate and multivariate COX proportional hazards analyses were applied to evaluate the associations of LMR with overall survival (OS, disease-free survival (DFS, distant metastasis-free survival (DMFS and loco-regional recurrence-free survival (LRRFS, respectively. RESULTS: Univariate analysis revealed that higher LMR level (≥ 5.220 was significantly associated with superior OS, DFS and DMFS (P values <0.001. The higher lymphocyte count (≥ 2.145 × 10(9/L was significantly associated with better OS (P = 0.002 and DMFS (P = 0.031, respectively, while the lower monocyte count (<0.475 × 10(9/L was associated with better OS (P = 0.012, DFS (P = 0.011 and DMFS (P = 0.003, respectively. Multivariate Cox proportional hazard analysis showed that higher LMR level was a significantly independent predictor for superior OS (hazard ratio or HR = 0.558, 95% confidence interval or 95% CI = 0.417-0.748; P<0.001, DFS (HR = 0.669, 95% CI = 0.535-0.838; P<0.001 and DMFS (HR = 0.543, 95% CI = 0.403-0.732; P<0.001, respectively. The advanced T and N stages were also independent indicators for worse OS, DFS, and DMFS, except that T stage showed borderline statistical significance for DFS (P = 0.053 and DMFS (P = 0.080. CONCLUSIONS: The

  9. 半滑舌鳎外周血淋巴细胞培养条件研究%Study on blood lymphocytic cell cultivation of half smooth tongue sole ( Cynoglossus semilaevis)

    Institute of Scientific and Technical Information of China (English)

    王美玉; 刘海金; 范兆廷; 宋立民

    2012-01-01

    以半滑舌鳎为试验对象,对淋巴细胞体外培养和染色体制备条件如促分裂剂、培养时间、培养温度、培养基、秋水仙素浓度及作用时间等进行探索,在建立一套基于半滑舌鳎淋巴细胞体外培养的染色体制备方法.结果表明,对于半滑舌鳎淋巴细胞体外培养,用离心法收集淋巴细胞,培养基为含15%体积分数胎牛血清的L-15培养基,添加3μg·mL-1的PHA-P作促分裂剂,分裂指数可达2.6%±0.2%,6d后收集细胞制作染色体,获得较好效果.%Half smooth tongue sole(Cynogtossus semilaevis) was used to explore the condition of the peripheral blood lymphocytic cell cultivation and chromosome preparation. The conditions were searched from mrtogens, incubation periods, culture temperature, media and the effect of the chloride. The aim was to establish a systemic method. The results showed that lymphocytes isolated from fresh blood by a stirring method were cultured in medium L-15 supplemented with 15%FBS, 3 μg·mL-1 PHA (phytohemagglutinin) as mitogens, and collected at day 6. This was the optimized culture condition for half smooth tongue sole. After culture to make chromosome, the mitotic index (Ml) could reach (2.6±0.2)%.

  10. Determination of TiO2, ZrO2, and Al2O3 nanoparticles on genotoxic responses in human peripheral blood lymphocytes and cultured embyronic kidney cells.

    Science.gov (United States)

    Demir, Eşref; Burgucu, Durmuş; Turna, Fatma; Aksakal, Sezgin; Kaya, Bülent

    2013-01-01

    In this study a genotoxic evaluation of titanium dioxide (TiO2, 2.3 nm), zirconium oxide (ZrO2, 6 nm), aluminum oxide (Al2O3, 16.7 nm) nanoparticles (NP) and their ionic forms was conducted using human peripheral blood lymphocytes and cultured human embryonic kidney (HEK293) cells by means of a modified alkaline comet assay with/without the formamidopyrimidine-DNA glycosylase (Fpg) and endonuclease III (Endo III) enzymes. Modifications to the comet assay by using lesion-specific endonucleases, such as Endo III and Fpg, detect DNA bases with oxidative damage. Both human peripheral blood lymphocytes and cultured embryonic kidney cells were incubated with TiO2, ZrO2, or Al2O3 NP at concentrations of 1, 10, or 100 μg/ml. Our results showed no significant induction in DNA damage by the comet assay with/without the Endo III and Fpg enzymes at all concentrations of ZrO2 and Al2O3. In the case of TiO2 NP only the highest concentration of 100 μg/ml significantly induced a genotoxic response. Data thus indicate that both ZrO2 and Al2O3 NP were not genotoxic in our system and in the case of TiO2 the lowest-observed-adverse-effect level (LOAEL) for genotoxicity was 100 μg/ml. Evidence indicates that these metallic NP are considered safe in light of the fact that no genotoxicity was noted with ZrO2 and Al2O3 and that the highest TiO2 concentration is not environmentally relevant.

  11. Study on B lymphocyte,T lymphocyte and its subgroups in peripheral blood in patients with hepatitis C%丙型肝炎患者外周血B细胞和T细胞及其亚群的研究

    Institute of Scientific and Technical Information of China (English)

    刘洪月; 何浩明; 田小平; 徐凤英; 庄惠芹; 袁兵

    2001-01-01

    Objective To study the changes of B lymphocyte,T lymphocyte,and its subgroups in peripheral blood in patients with hepatitis C.Methods B lymphocyte,T lymphocyte and its subgroups in peripheral blood in 38 patients with hepatitis C and 35 normal controls were detected with monoclonal technigue.Results The level of B lymphocyte were significanty higher than those in normal people(P<0.001).But CD3、CD4 and the ratio of CD4/CD8 were obviously lower(P<0.01,P<0.001).Conclusion Hepatitis C is a kind of selfi-mmunity disease with abnormal immunoregulation.%目的了解丙型肝炎患者外周血B细胞和T淋巴细胞及其亚群的变化。方法应用单克隆技术测定38例丙型肝炎患者外周血B细胞和T淋巴细胞亚群的水平并以35名健康人作对照。结果丙型肝炎患者外周血B细胞数显著地高于正常人(P<0.001)、CD3、CD4、CD4/CD8显著低于正常人(P<0.01,P<0.001)。结论丙型肝炎患者为一种自身免疫调节异常的疾病。

  12. Chronic lymphocytic leukaemia

    Science.gov (United States)

    Kipps, Thomas J.; Stevenson, Freda K.; Wu, Catherine J.; Croce, Carlo M.; Packham, Graham; Wierda, William G.; O’Brien, Susan; Gribben, John; Rai, Kanti

    2017-01-01

    Chronic lymphocytic leukaemia (CLL) is a malignancy of CD5+ B cells that is characterized by the accumulation of small, mature-appearing lymphocytes in the blood, marrow and lymphoid tissues. Signalling via surface immunoglobulin, which constitutes the major part of the B cell receptor, and several genetic alterations play a part in CLL pathogenesis, in addition to interactions between CLL cells and other cell types, such as stromal cells, T cells and nurse-like cells in the lymph nodes. The clinical progression of CLL is heterogeneous and ranges from patients who require treatment soon after diagnosis to others who do not require therapy for many years, if at all. Several factors, including the immunoglobulin heavy-chain variable region gene (IGHV) mutational status, genomic changes, patient age and the presence of comorbidities, should be considered when defining the optimal management strategies, which include chemotherapy, chemoimmunotherapy and/or drugs targeting B cell receptor signalling or inhibitors of apoptosis, such as BCL-2. Research on the biology of CLL has profoundly enhanced our ability to identify patients who are at higher risk for disease progression and our capacity to treat patients with drugs that selectively target distinctive phenotypic or physiological features of CLL. How these and other advances have shaped our current understanding and treatment of patients with CLL is the subject of this Primer. PMID:28102226

  13. HIV合并HBV/HCV感染患者外周血T细胞亚群的变迁%The changes of peripheral blood T lymphocyte subset in the patients of HIV concurrent infection with HBV/HCV

    Institute of Scientific and Technical Information of China (English)

    栗群英; 吴丽娟; 陈莉; 刘毓刚; 彭燕

    2011-01-01

    Objective To Analyze of the change character of peripheral blood T lymphocyte subset in the patients of HIV concurrent infection with HBV/HCV, and investigate the immune function of these patients. Methods Three color Flow cytometry was used to determine the peripheral blood total T cell, CD4 + T cell, CD8+T cell and calculate CD4 + T/ CD8+T in 26 patients of HIV concurrent infection with HBV/HCV(concurrent infection group), 35 patients with HIV only(simple infection group) and 194 health people(control group). Results The peripheral blood total T cell, CD4+T cell, CD8+T cell and CD4+T/ CD8+T in simple infection group were 66. 42 ± 4. 11,27. 74 ± 2. 34,47. 72 ± 3. 86 and 0. 74 ± 0. 19 ,or concurrent infection group were 54. 76 ± 3. 42, 22. 31 ± 1. 87, 58. 23 ± 4. 62 and 0. 33 ± 0. 12;and control group were 69. 98 ± 5. 79,35. 25 ± 5. 16,25. 08 ± 4. 34 and 1. 45 ± 0. 28. Significantly higher percentage of CD8+T lymphocyte, lower of CD4+T lymphocyte and CD4 + /CD8+ ratio in simple infection group were found , comparing to those in controls (P<0. 05 or P<0. 01). It was also demonstrate that in the group of concurrent infection, the percentage of total T lymphocyte, CD4 + T lymphocyte and CD4 + /CD8+ were lower and CD8+T lymphocyte was higher than those in simple infection group and control group (P<0. 05 or P<0. 01). Conclusion Concurrent infection with HBV and HCV in patient of HIV infection will aggravate immune disorder; Detection of peripheral blood T lymphocyte subset can be used to differential diagnosis, patients condition motion and prognosis judgment in patients of HIV/HBV/HCV multiple infection.%目的 了解HIV合并HBV、HCV感染患者外周血T细胞亚群的变化特征,探讨HIV合并HBV和HCV感染后患者的免疫功能.方法 采用三色流式细胞学检测技术,对26例HIV合并HBV和HCV感染患者(合并感染组)、35例单纯HIV感染患者(单纯感染组)和194例健康人群(健康对照组)外周血总T细胞、T4细胞、T8

  14. A new extract of the plant calendula officinalis produces a dual in vitro effect: cytotoxic anti-tumor activity and lymphocyte activation

    Directory of Open Access Journals (Sweden)

    Collado Antonia

    2006-05-01

    Full Text Available Abstract Background Phytopharmacological studies of different Calendula extracts have shown anti-inflamatory, anti-viral and anti-genotoxic properties of therapeutic interest. In this study, we evaluated the in vitro cytotoxic anti-tumor and immunomodulatory activities and in vivo anti-tumor effect of Laser Activated Calendula Extract (LACE, a novel extract of the plant Calendula Officinalis (Asteraceae. Methods An aqueous extract of Calendula Officinalis was obtained by a novel extraction method in order to measure its anti-tumor and immunomodulatory activities in vitro. Tumor cell lines derived from leukemias, melanomas, fibrosarcomas and cancers of breast, prostate, cervix, lung, pancreas and colorectal were used and tumor cell proliferation in vitro was measured by BrdU incorporation and viable cell count. Effect of LACE on human peripheral blood lymphocyte (PBL proliferation in vitro was also analyzed. Studies of cell cycle and apoptosis were performed in LACE-treated cells. In vivo anti-tumor activity was evaluated in nude mice bearing subcutaneously human Ando-2 melanoma cells. Results The LACE extract showed a potent in vitro inhibition of tumor cell proliferation when tested on a wide variety of human and murine tumor cell lines. The inhibition ranged from 70 to 100%. Mechanisms of inhibition were identified as cell cycle arrest in G0/G1 phase and Caspase-3-induced apoptosis. Interestingly, the same extract showed an opposite effect when tested on PBLs and NKL cell line, in which in vitro induction of proliferation and activation of these cells was observed. The intraperitoneal injection or oral administration of LACE extract in nude mice inhibits in vivo tumor growth of Ando-2 melanoma cells and prolongs the survival day of the mice. Conclusion These results indicate that LACE aqueous extract has two complementary activities in vitro with potential anti-tumor therapeutic effect: cytotoxic tumor cell activity and lymphocyte activation

  15. Calibration of the γ-H2AX DNA double strand break focus assay for internal radiation exposure of blood lymphocytes.

    Science.gov (United States)

    Eberlein, Uta; Peper, Michel; Fernández, Maria; Lassmann, Michael; Scherthan, Harry

    2015-01-01

    DNA double strand break (DSB) formation induced by ionizing radiation exposure is indicated by the DSB biomarkers γ-H2AX and 53BP1. Knowledge about DSB foci formation in-vitro after internal irradiation of whole blood samples with radionuclides in solution will help us to gain detailed insights about dose-response relationships in patients after molecular radiotherapy (MRT). Therefore, we studied the induction of radiation-induced co-localizing γ-H2AX and 53BP1 foci as surrogate markers for DSBs in-vitro, and correlated the obtained foci per cell values with the in-vitro absorbed doses to the blood for the two most frequently used radionuclides in MRT (I-131 and Lu-177). This approach led to an in-vitro calibration curve. Overall, 55 blood samples of three healthy volunteers were analyzed. For each experiment several vials containing a mixture of whole blood and radioactive solutions with different concentrations of isotonic NaCl-diluted radionuclides with known activities were prepared. Leukocytes were recovered by density centrifugation after incubation and constant blending for 1 h at 37°C. After ethanol fixation they were subjected to two-color immunofluorescence staining and the average frequencies of the co-localizing γ-H2AX and 53BP1 foci/nucleus were determined using a fluorescence microscope equipped with a red/green double band pass filter. The exact activity was determined in parallel in each blood sample by calibrated germanium detector measurements. The absorbed dose rates to the blood per nuclear disintegrations occurring in 1 ml of blood were calculated for both isotopes by a Monte Carlo simulation. The measured blood doses in our samples ranged from 6 to 95 mGy. A linear relationship was found between the number of DSB-marking foci/nucleus and the absorbed dose to the blood for both radionuclides studied. There were only minor nuclide-specific intra- and inter-subject deviations.

  16. Calibration of the γ-H2AX DNA double strand break focus assay for internal radiation exposure of blood lymphocytes.

    Directory of Open Access Journals (Sweden)

    Uta Eberlein

    Full Text Available DNA double strand break (DSB formation induced by ionizing radiation exposure is indicated by the DSB biomarkers γ-H2AX and 53BP1. Knowledge about DSB foci formation in-vitro after internal irradiation of whole blood samples with radionuclides in solution will help us to gain detailed insights about dose-response relationships in patients after molecular radiotherapy (MRT. Therefore, we studied the induction of radiation-induced co-localizing γ-H2AX and 53BP1 foci as surrogate markers for DSBs in-vitro, and correlated the obtained foci per cell values with the in-vitro absorbed doses to the blood for the two most frequently used radionuclides in MRT (I-131 and Lu-177. This approach led to an in-vitro calibration curve. Overall, 55 blood samples of three healthy volunteers were analyzed. For each experiment several vials containing a mixture of whole blood and radioactive solutions with different concentrations of isotonic NaCl-diluted radionuclides with known activities were prepared. Leukocytes were recovered by density centrifugation after incubation and constant blending for 1 h at 37°C. After ethanol fixation they were subjected to two-color immunofluorescence staining and the average frequencies of the co-localizing γ-H2AX and 53BP1 foci/nucleus were determined using a fluorescence microscope equipped with a red/green double band pass filter. The exact activity was determined in parallel in each blood sample by calibrated germanium detector measurements. The absorbed dose rates to the blood per nuclear disintegrations occurring in 1 ml of blood were calculated for both isotopes by a Monte Carlo simulation. The measured blood doses in our samples ranged from 6 to 95 mGy. A linear relationship was found between the number of DSB-marking foci/nucleus and the absorbed dose to the blood for both radionuclides studied. There were only minor nuclide-specific intra- and inter-subject deviations.

  17. Autologous Peripheral Blood Stem Cell Transplant Followed by Donor Bone Marrow Transplant in Treating Patients With High-Risk Hodgkin Lymphoma, Non-Hodgkin Lymphoma, Multiple Myeloma, or Chronic Lymphocytic Leukemia

    Science.gov (United States)

    2016-06-17

    B-Cell Prolymphocytic Leukemia; Plasma Cell Leukemia; Progression of Multiple Myeloma or Plasma Cell Leukemia; Recurrent Adult Hodgkin Lymphoma; Recurrent Adult Non-Hodgkin Lymphoma; Recurrent Childhood Hodgkin Lymphoma; Recurrent Childhood Non-Hodgkin Lymphoma; Recurrent Chronic Lymphocytic Leukemia; Recurrent Plasma Cell Myeloma; Recurrent Small Lymphocytic Lymphoma; Refractory Childhood Hodgkin Lymphoma; Refractory Chronic Lymphocytic Leukemia; Refractory Non-Hodgkin Lymphoma; Refractory Plasma Cell Myeloma; Refractory Small Lymphocytic Lymphoma; T-Cell Prolymphocytic Leukemia; Waldenstrom Macroglobulinemia

  18. The influence of x-ray contrast agents in computed tomography on the induction of dicentrics and {gamma}-H2AX foci in lymphocytes of human blood samples

    Energy Technology Data Exchange (ETDEWEB)

    Jost, G; Golfier, S; Pietsch, H; Lengsfeld, P; Voth, M [Bayer Schering Pharma AG, 13353 Berlin (Germany); Schmid, T E [Department of Radiation Oncology, Klinikum rechts der Isar, Technische Universitaet Muenchen, 81675 Munich (Germany); Eckardt-Schupp, F [Institute of Radiation Biology, Helmholtz Zentrum Muenchen, German Research Center for Environmental Health, 85764 Neuherberg (Germany); Schmid, E [Institute for Cell Biology, Center for Integrated Protein Science, University of Munich, 80336 Muenchen (Germany)], E-mail: Ernst.Schmid@lrz.uni-muenchen.de

    2009-10-21

    The aim of this study was to investigate and quantify two biomarkers for radiation exposure (dicentrics and {gamma}-H2AX foci) in human lymphocytes after CT scans in the presence of an iodinated contrast agent. Blood samples from a healthy donor were exposed to CT scans in the absence or presence of iotrolan 300 at iodine concentrations of 5 or 50 mg ml{sup -1} blood. The samples were exposed to 0.025, 0.05, 0.1 and 1 Gy in a tissue equivalent body phantom. Chromosome aberration scoring and automated microscopic analysis of {gamma}-H2AX foci were performed in parts of the same samples. The theoretical physical dose enhancement factor (DEF) was calculated on the basis of the mass energy-absorption coefficients of iodine and blood and the photon energy spectrum of the CT tube. No significant differences in the yields of dicentrics and {gamma}-H2AX foci were observed in the absence or presence of 5 mg iodine ml{sup -1} blood up to 0.1 Gy, whereas at 1 Gy the yields were elevated for both biomarkers. At an iodine concentration of 50 mg ml{sup -1} serving as a positive control, a biological DEF of 9.5 {+-} 1.4 and 2.3 {+-} 0.5 was determined for dicentrics and {gamma}-H2AX foci, respectively. A physical DEF of 1.56 and 6.30 was calculated for 5 and 50 mg iodine ml{sup -1}, respectively. Thus, it can be concluded that in the diagnostic dose range (radiation and contrast dose), no relevant biological dose-enhancing effect could be detected, whereas a clear biological dose-enhancing effect could be found for a contrast dose well outside the diagnostic CT range for the complete radiation dose range with both methods.

  19. An Elevated Peripheral Blood Monocyte-to-Lymphocyte Ratio Predicts Poor Prognosis in Patients with Primary Pulmonary Lymphoepithelioma-Like Carcinoma.

    Directory of Open Access Journals (Sweden)

    Liang Wang

    Full Text Available Primary pulmonary lymphoepithelioma-like carcinoma (LELC is a rare type of non-small cell lung cancer. In this study, we retrospectively reviewed the data from 74 consecutive patients with pulmonary LELC and investigated the prognostic value of pretreatment monocyte-to-lymphocyte ratio (MLR. The cut-off value determined by ROC curve for MLR was 0.262. According to this cut-off value, 36 (48.6% patients had lower MLR value ( = 0.262 were significantly associated with poor OS and PFS. In a multivariate Cox regression model that included stage, LDH and MLR level, all of these three factors were found to be independent prognostic factors for both PFS and OS. In patients who received radical surgery, MLR level remained significantly correlated with OS and PFS. In conclusion, we firstly demonstrated that pretreatment MLR can be used as a useful independent prognostic marker in patients with pulmonary LELC, and might guide us to optimize the treatment strategies. However, due to the relatively rarity of this disease and the limitation of a retrospective study, further prospective studies performed in multicenter are necessary to validate the prognostic value of MLR in pulmonary LELC.

  20. Relationship between T lymphocytes counts and peripheral blood components in HIV/AIDS patients%HIV/AIDS患者T淋巴细胞计数与外周血组份间关系的研究

    Institute of Scientific and Technical Information of China (English)

    贾海英; 陈莉霞; 马慧霞; 米吉提.买买提

    2012-01-01

    Objective To explore the relationship between peripheral T lymphocyte count and blood test results among HIV/AIDS patients. Methods Anticoagulant were collected from 78 cases of HIV/AIDS (HIV/AIDS group) and 55 cases of healthy volunteers (healthy control group), CD4+T and CD8+T lymphocytes in peripheral blood by flow cytometry were detected , while detected blood cells routine testing were by automatic hematology analyzer.Results 71.79% (56/78) in 78 patients CD4+ T lymphocyte count were <200 cells /μl, 98.72% patients CD47CD8Vere<0.8,among which 75.64% (59/78) patients CD4/CD8 between 0.1~0.5. Those six projects CD4+,CD8+, CD3+,CD4/CD8,WBC and LYMPH both had statistics significance when HIV/ AIDS group compared with control group. While those three projects NEUT,% NEUT and % LYMPH had no statistics significance. Conclusion It is closely related to CD4+,CD8+, CD3+, CD4+CD8+ and LYMPH in the disease process of HIV/ AIDS patients. CD4+,CD8+, blood cells routine testing are detected early to understand the progression of the disease and antiviral therapy time.%目的 分析HIV/AIDS患者外周血T淋巴细胞计数和血常规检测结果.方法 收集78例HIV/AIDS患者(HIV/AIDS组)、55例健康体检人员(健康对照组)的抗凝血,用流式细胞仪技术检测外周血CD4+T及CD8+T淋巴细胞,全自动血细胞分析仪检测血常规.结果 78例患者中有71.79%(56/78) CD4+T淋巴细胞数<200个/μl,98.72%的HIV/AIDS患者CD4+/CD8+比值<0.8,其中75.64% (59/78)患者4/8介于0.1~0.5之间.CD4+、CD8+、CD3+、CD4/CD8、WBC、LYMPH六个项目HIV/AIDS组与健康对照组比较差异均有统计学意义(P<0.05),NEUT、%NEUT及%LYMPH三个项目两组间比较差异无统计学意义(P>0.05).结论 HIV/AIDS患者在发病过程中与CD4+、CD8+、CD3+、CD4+/CD8+及LYMPH密切相关,建议HIV感染者尽早进行医学咨询并做CD4+、CD8+和血常规的检测,以了解病情的进展及进行抗病毒治疗.

  1. The correlation of the liver damage induced by Epstein-Barr virus infection with peripheral blood lymphocyte subsets%EB病毒感染肝脏损伤与外周血淋巴细胞亚群相关性

    Institute of Scientific and Technical Information of China (English)

    张慧; 李双杰; 袁远宏; 欧阳文献; 康桢

    2013-01-01

    目的 了解EB病毒(EBV)感染患儿外周血游离EBV-DNA拷贝数,淋巴细胞分化抗原表达率与EBV感染相关性肝炎的关系.方法 应用荧光定量PCR方法,测定46例EBV感染肝功能正常患儿,43例EBV感染肝功能异常患儿外周血血浆中游离EBV-DNA拷贝数;利用流式细胞仪测定EBV感染患儿CD4+、CD8+、CD4+/CD8+、CD16+56(NK)表达率并与32例正常对照儿童进行比较.结果 EBV感染肝功能正常组与肝功能异常组患儿比较,血浆中游离EBV-DNA水平差异无统计学意义(P=0.24).EBV感染肝功能正常组、EBV感染肝功能异常组以及正常对照儿童的总T细胞、CD4+、CD4+/CD8+差异有统计学意义(P均<0.05).EBV感染肝功能异常组患儿的CD4+、CD4+/CD8+较EB感染肝功能正常组下降,差异有统计学意义(P均<0.05);两组EBV感染患儿与正常对照儿童比较,总T细胞、CD4+、CD4+/CD8+降低,CD8+增高,差异有统计学意义(P均<0.00).结论 EBV感染肝脏损伤及严重程度与外周血淋巴细胞分化抗原中T细胞亚群有一定相关性.%Objective To measure the cell-free Epstein-Barr virus (EBV) DNA copy number in peripheral blood plasma of of children with EBV infection, and to study the relationship of the expression rates of peripheral blood lymphocyte differentiation antigens with EBV infection associated hepatitis. Methods The cell-free EBV DNA copy number in peripheral blood plasma was determined by fluorescence quantitative PCR in 46 EBV infection children with normal liver function and 43 EBV infection children with abnormal liver function. The expression rates of lymphocyte differentiation antigens of CD4+, CD8+, CD4+/CD8+ and CD16+56 (NK) were determined by flow cytometry and compared between EBV infection children and 32 normal control children. Results There was no significant difference in the cell-free EBV DNA copy number between EBV infection children with normal or abnormal liver function (P=0.24). There was significant difference

  2. Evaluation of genotoxic and cytotoxic effects of {sup 153} Sm-EDTMP in peripheral blood lymphocytes of bone metastasis patients; Avaliacao dos efeitos genotoxico e citotoxico do {sup 153} Sm-EDTMP em linfocitos perifericos de pacientes com metastase ossea

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, Miriam Fussae

    2003-07-01

    In this study the cellular damage in peripheral lymphocytes after exposure to {sup 153} Sm-EDTMP (Samarium-153 ethylene-diamine-tetramietylene-phosphonate) was determined using the technique of micronuclei analysis and differential coloration.{sup 153} Sm-EDTMP is a radiopharmaceutical used for pain relief in patients with bone metastases. The analysis of the frequency of micronuclei in patient blood samples obtained one hour after endovenous administration of radiopharmaceutical (41 MBq/kg) showed no statistical difference in relation to basal values in binucleated cells. However the analysis of damage distribution in mononucleated cells, showed that the patients without previous radiotherapy treatment presented a significant increase in the frequency of cells with one micronucleus and in those who had taken previous radiotherapy treatment, in cells with two or more micronuclei. The in vitro experiments conducted with the exposition of total blood to three radiation concentrations of {sup 153} Sm-EDTMP (0.370, 0.555 and 1.110 MBq/mL) during one hour showed an increase in the frequency of micronuclei and necrotic and apoptotic cells with increasing radiation dose. Dose-response curves for healthy donors and patients with bone metastasis without previous radiotherapy treatment were constructed. The comparison of the curves showed that patients presented higher radiosensitivity, either micronuclei or dead cell (necrotic or apoptotic) percentages, than healthy donors. (author)

  3. Caspase-3-mediated cleavage of p65/RelA results in a carboxy-terminal fragment that inhibits IκBα and enhances HIV-1 replication in human T lymphocytes

    Directory of Open Access Journals (Sweden)

    Alcamí José

    2008-12-01

    Full Text Available Abstract Background Degradation of p65/RelA has been involved in both the inhibition of NF-κB-dependent activity and the onset of apoptosis. However, the mechanisms of NF-κB degradation are unclear and can vary depending on the cell type. Cleavage of p65/RelA can produce an amino-terminal fragment that was shown to act as a dominant-negative inhibitor of NF-κB, thereby promoting apoptosis. However, the opposite situation has also been described and the production of a carboxy-terminal fragment that contains two potent transactivation domains has also been related to the onset of apoptosis. In this context, a carboxy-terminal fragment of p65/RelA (ΔNH2p65, detected in non-apoptotic human T lymphocytes upon activation, has been studied. T cells constitute one of the long-lived cellular reservoirs of the human immunodeficiency virus type 1 (HIV-1. Because NF-κB is the most important inducible element involved in initiation of HIV-1 transcription, an adequate control of NF-κB response is of paramount importance for both T cell survival and viral spread. Its major inhibitor IκBα constitutes a master terminator of NF-κB response that is complemented by degradation of p65/RelA. Results and conclusions In this study, the function of a caspase-3-mediated carboxy-terminal fragment of p65/RelA, which was detected in activated human peripheral blood lymphocytes (PBLs, was analyzed. Cells producing this truncated p65/RelA did not undergo apoptosis but showed a high viability, in spite of caspase-3 activation. ΔNH2p65 lacked most of DNA-binding domain but retained the dimerization domain, NLS and transactivation domains. Consequently, it could translocate to the nucleus, associate with NF-κB1/p50 and IκBα, but could not bind -κB consensus sites. However, although ΔNH2p65 lacked transcriptional activity by itself, it could increase NF-κB activity in a dose-dependent manner by hijacking IκBα. Thus, its expression resulted in a persistent

  4. Rearrangements in human chromosome 1 visualized by arm-specific probes in the progeny of blood lymphocytes exposed to iron ions

    Science.gov (United States)

    Manti, L.; Bertucci, A.; Gialanella, G.; Grossi, G.; Pugliese, M.; Scampoli, P.; Durante, M.

    It is well known that heavy ions are more effective than sparsely ionizing radiation in the induction of chromosomal aberrations in heavy ions However most of the complex rearrangements induced by densely ionizing radiation ultimately lead to cell death For risk assessment it is more important to measure the residual cytotgenetic damage in cells surviving the exposure and able to proliferate This analyses will be strongly influenced by the technique used to visualize the chromosomes and consequently on the aberrations scored For instance symmetrical exchanges have higher transmission probability than asymmetrical exchanges Multi-fuor FISH including mBAND mFISH or RxFISH allows the detection of many different aberrations with high resolution but it is slow and expensive thus affecting the statistical power of the study In this study we hybridized metaphase cells with human DNA probes specific for the p and q arms of the chromosome 1 The arm-specific probes allow a fast and reliable detection of both symmetrical and asymmetrical inter-chromosomal exchanges and inter-arm intra-changes in the painted chromosome pair We used this method to score aberrations in human lymphocytes exposed to 1 Gy of either 250 kVp X-rays or 1 GeV n Fe-ions LET 145 keV mu m and harvested following 120 h in culture including 2 h in colcemid for metaphase-block Although iron ions are much more effective than X-rays in the induction of chromosomal aberrations formed during the first post-exposure cell-cycle we found that the effectiveness drops when

  5. Chromosome aberrations induced in human lymphocytes by U-235 fission neutrons: I. Irradiation of human blood samples in the "dry cell" of the TRIGA Mark II nuclear reactor.

    Science.gov (United States)

    Fajgelj, A; Lakoski, A; Horvat, D; Remec, I; Skrk, J; Stegnar, P

    1991-11-01

    A set-up for irradiation of biological samples in the TRIGA Mark II research reactor in Ljubljana is described. Threshold activation detectors were used for characterisation of the neutron flux, and the accompanying gamma dose was measured by TLDs. Human peripheral blood samples were irradiated "in vitro" and biological effects evaluated according to the unstable chromosomal aberrations induced. Biological effects of two types of cultivation of irradiated blood samples, the first immediately after irradiation and the second after 96 h storage, were studied. A significant difference in the incidence of chromosomal aberrations between these two types of samples was obtained, while our dose-response curve fitting coefficients alpha 1 = (7.71 +/- 0.09) x 10(-2) Gy-1 (immediate cultivation) and alpha 2 = (11.03 +/- 0.08) x 10(-2) Gy-1 (96 h delayed cultivation) are in both cases lower than could be found in the literature.

  6. Calibration Curve for Dicentric Chromosomes Induced in Human Blood Lymphocytes Exposed to Gamma Rays at a Dose Rate of 12.5 mGy/s.

    Science.gov (United States)

    Que, Tran; Duy, Pham Ngoc; Luyen, Bui Thi Kim

    2016-01-01

    To develop a calibration curve for induction of dicentric chromosomes by radiation, we have used a 60Co gamma-ray source with dose rate of 12.5 mGy/s. Whole blood from 15 healthy donors was collected. Whole blood from each donor was divided equally into 8 parts for exposing to supposedly physical doses 0, 0.30, 0.50, 1.00, 1.50, 2.00, 3.00 and 4.00 Gy for a independent calibration curve. Whole blood from 15 donors was used to calibrate dose - effect and statistical for general calibration curve. Using Poisson test (U-test) for the distribution of dicentric chromosomes in the metaphases to determine the uniformity of the radiation field. The average from 15 independent calibration curves of linear correlated coefficient was determined to be r (y, d) = 0.5136 ± 0.0038. The model equation derived is y = aD + bD(2) + C. The calibration equation of dose-effect was y = 1.01D + 4.43D(2) + 0.56.

  7. 外周血淋巴细胞染色体G显带技术的改良%Improvement of G banding technique for chromosome of peripheral blood lymphocytes

    Institute of Scientific and Technical Information of China (English)

    莫凤明; 杨丽华; 张璐

    2013-01-01

    目的 探讨外周血淋巴细胞染色体吉姆萨(Giemsa)显带技术(G显带技术)的改良,提高G显带效果,并能节约时间,进一步提高染色体诊断效率.方法 250例疑似患者用常规方法进行外周血淋巴细胞培养及制片、烤片,用0.05%胰蛋白酶工作液消化20 s,Giemsa染液染色,并与0.025%胰蛋白酶工作液消化时间5~6 min的常规显带方法对比.结果 改良方法的标本片带纹清晰,深浅带反差突出,合格率98%,最佳率72.8%,而常规方法合格率98%,最佳率44.8%,2种方法在得到最佳片数上差异有统计学意义(P<0.01).结论 应用改良技术可缩短染色体制片时间,又能达到最佳效果,同时为工作人员节约时间,及早出结果,可提高临床对染色体病诊断效率,因此本改良方法是值得推广的实验方法.%Objective To investigate the improvement of Giemsa banding technique (G banding technique) for chromosome of peripheral blood lymphocytes,and improve the effects of G banding,save time and further improve the efficiency in the diagnosis of chromosome.Methods For 250 suspected patients,cultivation and smear production of peripheral blood lymphocytes were carried out by routine method.Peripheral blood lymphocytes were digested in 0.05%trypsin working solution for 20s,and stained by Giemsa.The results were compared with the routine banding technique digested in 0.025% trypsin working solution for 5-6 min.Results Specimnen by improved method presented clear band and repeated prominent shade band,with 98% of pass and 72.8% of optimal ratio,while the routine method achieved 98% of pass and 44.8% of optimal ratio.Two methods had statistical significance in optimal ratio (P < 0.01).Conclusions The application of improved technique not only shortens time for smear production of chromosome and achieves optimal effects,but also saves time for staffs,and shows results as soon as possible,improving efficiency in clinical

  8. 儿童再生障碍性贫血淋巴细胞亚群及与血常规相关性的研究%Lymphocyte subsets and its correlation with blood count in children with aplastic anemia

    Institute of Scientific and Technical Information of China (English)

    曹萍; 蒋莎义; 谢晓恬; 周妮娜; 李福兴; 覃大卫

    2011-01-01

    Objective To investigate the distribution of lymphocyte subsets in peripheral blood and its correlation with complete blood count in children with aplastic anemia. Methods The lymphocyte subsets CD3+ , CD3 + CD4+ , CD3 + CD8 + , CD3+ CD4 VCD3 + CD8 + , CD3 + CD45RA+, CD3 + CD45RO+ , CD(16 +56) + ,CD19 +from 42 aplastic anemia children and 27 controls were examined by flow cytometry; its correlations with complete blood count were analyzed. Results CD3 + CD4+ and CD3 + CD4 VCD3 + CD8+ decreased while CD3 + CD8+ increased in aplastic anemia patients. CD3 + CD8+ was positively correlated with peripheral HB, WBC counts. CD3+ CD4VCD3+ CD8+ was negatively correlated with peripheral HB, WBC and N counts. Conclusion There are abnormal distributions of lymphocyte subsets in aplastic anemia patients, which may be involved in pathogenesisof the disease.%目的 研究再生障碍性贫血患儿外周血淋巴细胞亚群分布,及其与同期外周血血常规的相关性.方法 采用流式细胞技术对我院42例AA患儿和27例正常儿童外周血淋巴细胞亚群CD3+,CD3+ CD4+,CD3+CD8+,CD3+ CD4+/CD3+ CD8+,CD3+ CD45 RA+,CD3+ CD45RO+,CD( 16 +56)+,CD19+的表达进行检测,同时检测血常规.结果 再障组CD3+ CD4+,CD3+ CD8+,CD3+ CD4+/CD3+ CD8+的平均值分别为(26.94±11.93)%,(33.33±10.3)%,0.918±0.53,对照组分别为(33.53±9.97)%,(27.08±9.23)%,1.44±0.80,差异有显著性(P<0.05);CD3+ CD8+与HB,WBC呈一定程度的正相关,CD3+ CD4+/CD3+ CD8+比值与HB,WBC,N呈一定程度的负相关.结论 再障患儿存在淋巴细胞亚群的失调,其中由T淋巴细胞介导的免疫异常对造血功能的抑制在再障的发生中起重要作用,再障患儿Th格局向Th1偏移.

  9. Cytotoxicity of lymphocytes from melanoma patients against autologous tumor cells and its potentiation in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Bykovskaya, S.N.; Iobadze, M.S.; Kupriyanova, T.A.; Demidov, L.V.

    1987-06-01

    The specific and natural cytotoxicity of peripheral blood lymphocytes from patients with melanomas was compared and stimulation with autologous tumor cells or a pool of allogeneic lymphocytes from five healthy blood donors also was used to potentiate the specific antitumor activity of the patients' lymphocytes. To assess cytolytic ability, cells of an autologous tumor, cells of the K-562 line, autologous peripheral blood lymphocytes, and blast cells obtained from these lymphocytes after stimulation by phytohemagglutinin were used as the target cells. The target cells were incubated in a medium containing sodium chromate and were labelled with the chromium 51 isotope.

  10. Role of oxidative stress and intracellular calcium in nickel carbonate hydroxide-induced sister-chromatid exchange, and alterations in replication index and mitotic index in cultured human peripheral blood lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    M' Bemba-Meka, Prosper [Universite de Montreal, Human Toxicology Research Group (TOXHUM), Department of Environmental and Occupational Health, Main Station, P.O. Box 6128, Montreal, QC (Canada); University of Louisville, Department of Pharmacology and Toxicology, Center for Genetics and Molecular Medicine, Louisville, KY (United States); Lemieux, Nicole [Universite de Montreal, Department of Pathology and Cellular Biology, Faculty of Medicine, Main Station, P.O. Box 6128, Montreal, QC (Canada); Chakrabarti, Saroj K. [Universite de Montreal, Human Toxicology Research Group (TOXHUM), Department of Environmental and Occupational Health, Main Station, P.O. Box 6128, Montreal, QC (Canada)

    2007-02-15

    Human peripheral lymphocytes from whole blood cultures were exposed to either soluble form of nickel carbonate hydroxide (NiCH) (0-60 {mu}M), or of nickel subsulfide (Ni{sub 3}S{sub 2}) (0-120 {mu}M), or of nickel oxide (NiO) (0-120 {mu}M), or nickel sulfate (NiSO{sub 4}) (0-120 {mu}M) for a short duration of 2 h. The treatments occurred 46 h after the beginning of the cultures. The cultures were harvested after a total incubation of 72 h, and sister-chromatid exchange (SCE), replication index (RI), and mitotic index (MI) were measured for each nickel compound. The soluble form of NiCH at 30 {mu}M but those of Ni{sub 3}S{sub 2} and NiO at 120 {mu}M produced significant increase in the SCE per cell compared to the control value, whereas NiSO{sub 4} failed to produce any such significant increase. Except NiSO{sub 4}, the soluble forms of NiCH, Ni{sub 3}S{sub 2}, and NiO produced significant cell-cycle delay (as measured by the inhibition of RI) as well as significant inhibition of the MI at respective similar concentrations as mentioned above. Pretreatment of human blood lymphocytes with catalase (H{sub 2}O{sub 2} scavenger), or superoxide dismutase (superoxide anion scavenger), or dimethylthiourea (hydroxyl radical scavenger), or deferoxamine (iron chelator), or N-acetylcysteine (general antioxidant) inhibited NiCH-induced SCE, and changes in RI and MI. This suggests the participation of oxidative stress involving H{sub 2}O{sub 2}, the superoxide anion radical, the hydroxyl radical, and iron in the NiCH-induced genotoxic responses. Cotreatment of NiCH with either verapamil (inhibitor of intracellular calcium ion ([Ca{sup 2+}]{sub i}) movement through plasma membranes), or dantrolene (inhibitor of [Ca{sup 2+}]{sub i} release from sarcoplasmic reticulum), or BAPTA (Ca{sup 2+} chelator) also inhibited the NiCH-induced responses. These results suggest that [Ca{sup 2+}]{sub i} is also implicated in the genotoxicity of NiCH. Overall these data indicate that various types

  11. Distinct phenotype, longitudinal changes of numbers and cell-associated virus in blood dendritic cells in SIV-infected CD8-lymphocyte depleted macaques.

    Science.gov (United States)

    Soulas, Caroline; Autissier, Patrick J; Burdo, Tricia H; Piatak, Michael; Lifson, Jeffrey D; Williams, Kenneth C

    2015-01-01

    Loss of circulating CD123+ plasmacytoid dendritic cells (pDCs) during HIV infection is well established. However, changes of myeloid DCs (mDCs) are ambiguous since they are studied as a homogeneous CD11c+ population despite phenotypic and functional heterogeneity. Heterogeneity of CD11c+ mDCs in primates is poorly described in HIV and SIV infection. Using multiparametric flow cytometry, we monitored longitudinally cell number and cell-associated virus of CD123+ pDCs and non-overlapping subsets of CD1c+ and CD16+ mDCs in SIV-infected CD8-depleted rhesus macaques. The numbers of all three DC subsets were significantly decreased by 8 days post-infection. Whereas CD123+ pDCs were persistently depleted, numbers of CD1c+ and CD16+ mDCs rebounded. Numbers of CD1c+ mDCs significantly increased by 3 weeks post-infection while numbers of CD16+ mDCs remained closer to pre-infection levels. We found similar changes in the numbers of all three DC subsets in CD8 depleted animals as we found in animals that were SIV infected animals that were not CD8 lymphocyte depleted. CD16+ mDCs and CD123+ pDCs but not CD1c+ mDCs were significantly decreased terminally with AIDS. All DC subsets harbored SIV RNA as early as 8 days and then throughout infection. However, SIV DNA was only detected in CD123+ pDCs and only at 40 days post-infection consistent with SIV RNA, at least in mDCs, being surface-bound. Altogether our data demonstrate that SIV infection differently affects CD1c+ and CD16+ mDCs where CD16+ but not CD1c+ mDCs are depleted and might be differentially regulated in terminal AIDS. Finally, our data underline the importance of studying CD1c+ and CD16+ mDCs as discrete populations, and not as total CD11c+ mDCs.

  12. Detailed analysis of immunologic effects of the cytotoxic T lymphocyte-associated antigen 4-blocking monoclonal antibody tremelimumab in peripheral blood of patients with melanoma

    Directory of Open Access Journals (Sweden)

    Gualberto Antonio

    2008-05-01

    Full Text Available Abstract Background CTLA4-blocking antibodies induce tumor regression in a subset of patients with melanoma. Analysis of immune parameters in peripheral blood may help define how responses are mediated. Methods Peripheral blood from HLA-A*0201-positive patients with advanced melanoma receiving tremelimumab (formerly CP-675,206 at 10 mg/kg monthly was repeatedly sampled during the first 4 cycles. Samples were analyzed by 1 tetramer and ELISPOT assays for reactivity to CMV, EBV, MART1, gp100, and tyrosinase; 2 activation HLA-DR and memory CD45RO markers on CD4+/CD8+ cells; and 3 real-time quantitative PCR of mRNA for FoxP3 transcription factor, preferentially expressed by T regulatory cells. The primary endpoint was difference in MART1-specific T cells by tetramer assay. Immunological data were explored for significant trends using clustering analysis. Results Three of 12 patients eligible for immune monitoring had tumor regression lasting > 2 years without relapse. There was no significant change in percent of MART1-specific T cells by tetramer assay. Additionally, there was no generalized trend toward postdosing changes in other antigen-specific CD8+ cell populations, FoxP3 transcripts, or overall changes in surface expression of T-cell activation or memory markers. Unsupervised hierarchical clustering based on immune monitoring data segregated patients randomly. However, clustering according to T-cell activation or memory markers separated patients with clinical response and most patients with inflammatory toxicity into a common subgroup. Conclusion Administration of CTLA4-blocking antibody tremelimumab to patients with advanced melanoma results in a subset of patients with long-lived tumor responses. T-cell activation and memory markers served as the only readout of the pharmacodynamic effects of this antibody in peripheral blood. Clinical trial registration number NCT00086489

  13. Peripheral blood lymphocytes micronucleus analysis of 450 hospital radiation workers in Qingdao area%青岛地区450名医院放射工作人员外周血淋巴细胞微核分析

    Institute of Scientific and Technical Information of China (English)

    孙冰梅; 王淑惠; 宋学术

    2014-01-01

    To investigate the statistical variance of micronucleus of peripheral blood lymphocytes between radiation group workers and control group workers in Qingdao area,and provide scientific evidence for professional health protection.Methods:The conventional culture method for lymphocytes and x2 test were used to analyze the micronucleus rates and micronucleated lymphocyte rates between 450 radiation group workers and 70 control group workers,between males and females,between radiodiagnosis,radiotherapy, interventional therapy and nuclear medicine.Results: The micronucleus rates and micronucleated lymphocyte rates in radiation group workers were higher than control group workers.The difference was significant (x2* =3.8809 x2**=3.9286 P<0.05).No significant difference was found between males and females,also between radiodiagnosis,radiotherapy, interventional therapy and nuclear medicine.Conclusions:Radiation group workers in hospital receive a long low-dose of ionizing radiation damage higher than natural background.This radiation damage can cause genetic material change. Effective safeguard measures could reduce the radiation damage.%目的:为了解青岛地区医院放射工作人员的淋巴细胞微核率与非接触放射线健康人群的微核率是否有统计学差异,为放射工作人员的辐射防护提供依据。方法:采用外周血淋巴细胞培养法进行微核检测,用x2检验对青岛地区450名医院放射工作人员和70名非接触放射线人员、男女性别及放射诊断、放射治疗、介入治疗和核医学四个工种之间比较进行统计分析。结果:医院从事放射工作人员的淋巴细胞微核率和微核细胞率显著高于对照组,差异均有统计学意义(x2值分别为3.8809,3.9286; P<0.05)。男女性别及从事放射诊断、放射治疗、介入治疗和核医学工作人员的淋巴细胞微核率和微核细胞率之间的差别均无统计学意义。结论:医院放射工作人

  14. Lymphocytes and liver fibrosis in HIV & HCV coinfection

    NARCIS (Netherlands)

    Feuth, M.

    2014-01-01

    Coinfection with HIV has an important impact on immunity against hepatitis C virus (HCV). In the present dissertation, phenotypes of lymphocytes derived from the peripheral blood of HCV-infected patients were studied into detail, with special attention to changes in phenotype of lymphocytes associat

  15. Carotenoid levels in human lymphocytes, measured by Raman microspectroscopy

    NARCIS (Netherlands)

    Ramanauskaite, R B; SegersNolten, IGMJ; DeGrauw, K J; Sijtsema, N M; VanderMaas, L; Greve, J; Otto, C; Figdor, C G

    1997-01-01

    Carotenoid levels in lymphocytes obtained from peripheral blood of healthy people have been investigated by Raman microspectroscopy. We observed that carotenoids are concentrated in so-called ''Gall bodies''. The level of carotenoids in living human lymphocytes was found to be age-dependent and to d

  16. Study on the Cytotoxic T Lymphocytes Clone Specific for the Nucleocapsid Protein of Hantaan Virus from Peripheral Blood in Patients with Hemorrhagic Fever with Renal Syndrome

    Institute of Scientific and Technical Information of China (English)

    潘蕾; 白雪帆; 黄长形; 李光玉

    2003-01-01

    In order to elucidate the molecular and immunological mechanisms as well as the pathogenesis of hemorrhagic fever with renal syndrome (HFRS), the CD8+ cytotoxic T lymphecytes (CTL) clone was established directly from peripheral blood mononuclear cells (PBMC) of patients with HFRS. The activities of CTL were detected as usual with EBV-transformed lymphoblastoid cell line (BLCL) as target cells. The results showed that the CTL clone could recognized and killed the targetcells with specificity of nucleocapsid protein of Hantaan virus (HTNVNP) with the cytotoxicity percentages of 50.2%,25.4% and 39.0% respectively. These results demonstrated that the antigenic epitopes of HTNVNP mainly located on the C-temainal of the viral nucleocapsid protein.

  17. [CHANGING OF PHYSICO-CHEMICAL PARAMETERS OF NON-CONTACT (ELECTROCHEMICAL) ACTIVATED DRINKING WATER IS ASSOCIATED WITH INDUCTION OF GENOMIC INSTABILITY OF CULTIVATED HUMAN BLOOD LYMPHOCYTES].

    Science.gov (United States)

    Zatsepina, O V; Ingel, F I

    2016-01-01

    In the article there are presented data which are the fragment of large multidisciplinary study of genetic safety of non-contact electrochemically activated water (NAW). The aim of this study was the analysis of the relation of impacts of genomic instability (micronucleus test with cytochalasin B) detected in human blood cells, cultured in medias prepared on the base of these NAWs, with physical and chemical properties of these NaWs. In experiments there were used catholytes and anolytes obtained by activation of osmotic, tap and dining bottled water As a result of such activation, all waters were shown to acquire the ability to induce genomic instability in cellular cultures. Notably in cell cultures on catholytes and anolytes these effects differed between themselves and have been associated with different physical and chemical properties of the NAWs.

  18. 江西省某铀矿工人周围血淋巴细胞染色体畸变分析%Analysis on chromosome aberration in peripheral blood lymphocytes among workers in a uranium mine of Jiangxi province

    Institute of Scientific and Technical Information of China (English)

    李小亮; 吕玉民; 韩林; 阮健磊; 陈以水; 孙全富; 刘建香

    2015-01-01

    目的:了解铀矿工人周围血淋巴细胞染色体畸变情况。方法采用整群随机抽样方法,选择江西省某铀矿98名铀矿工人为矿工组,48名接受放射工作人员培训但还未上岗的人员或行政工作人员为对照组,采集2组人员肘静脉血,采用培养开始加秋水仙素法培养50 h后收获细胞,常规方法制片,吉姆萨染液染色,显微镜下分析染色体畸变。结果矿工组染色体总畸变率为2.70%,对照组为2.49%,2组比较差异无统计学意义( Z=0.74,P=0.459)。矿工组“双着丝粒体+着丝粒环”(以下简称“双+环”)率为(0.25±0.05)%,对照组为(0.12±0.05)%。调整性别、年龄、工龄、吸烟和饮酒等因素后进行负二项回归分析,矿工组“双+环”率为对照组的5.76倍( Z=2.28,P=0.023)。结论铀矿工人的周围血淋巴细胞染色体非稳定性畸变水平较高,需要加强铀矿工人的职业性放射性疾病危害防治工作。%Objective To analyze the chromosome aberration of peripheral blood lymphocytes among uranium workers. Methods According to cluster sampling totally 98 workers from a uranium mine in Jiangxi province were randomly selected as the miner group.A total of 48 new employees who had received radiation pre-job training before going on duty or executive staff were selected as the control group.Their venous blood samples were collected.After 50 hours of cultivation by using colchicines at the beginning of cultivation method, blood cells were harvested and made into slides according to the conventional method.The slides were stained with Giemsa and chromosome aberrations were analyzed with microscope.Results The total aberration rate was 2.70%in the miner group and was 2.49%in the control group.There was no statistical significant difference between the two groups ( Z=0.74, P=0.459) .The rate of“dicentric and centric ring”(“dic+r”) was ( 0.25 ±0

  19. T- and B-lymphocyte chimerism in the marmoset

    Energy Technology Data Exchange (ETDEWEB)

    Niblack, G.D.; Kateley, J.R.; Gengozian, N.

    1977-01-01

    Marmosets are natural blood chimeras, this condition resulting from the high frequency of fraternal twinning and the consistent development of placental vascular anastomoses between the two embryos. Identification of chimerism by sex-chromosome analysis of cultured blood lymphocytes provided a means of determining the proportion of chimerism among T and B lymphocytes. Peripheral blood lymphocytes were enriched for T or B cells by filtration through a nylon column (yields >95% T-cells) or inactivation of T lymphocytes by treatment with a goat anti-marmoset thymocyte antiserum in the presence of complement (yields >95% B cells). Mitogenic stimulation of these separated, enriched cell populations yielded metaphase plates which could be scored for percentage male and female cells. Tests on five different blood chimeras showed the T- and B-lymphocyte chimerism to be the same. Stimulation of blood lymphocytes with cells from another species of marmoset in a mixed lymphocyte culture test revealed the chimeric T-cell response (i.e., host and co-twin cells) to be similar to that obtained with a mitogenic lectin. The demonstration of equivalent T- and B-cell chimerism in these animals suggests derivation of these cells from a common stem cell pool and the response of both T-cell populations to an antigenic stimulus in proportions similar to their percentage chimerism suggests complete immunologic tolerance exists in this species for co-twin histocompatibility antigens.

  20. Hyporesponsiveness of peripheral blood lymphocytes to streptococcal superantigens in patients with guttate psoriasis: evidence for systemic stimulation of T cells with superantigens released from focally infecting Streptococcus pyogenes.

    Science.gov (United States)

    Tokura, Y; Seo, N; Ohshima, A; Wakita, H; Yokote, R; Furukawa, F; Takigawa, M

    1999-01-01

    Throat infection with Streptococcus pyogenes is the most important trigger for acute guttate psoriasis. We examined the in vitro responses of peripheral blood mononuclear cells (PBMC) to streptococcal superantigens, SPEA and SPEC, and staphylococcal superantigens, SEB and TSST-1, in patients with guttate psoriasis, in patients with chronic plaque psoriasis, and in healthy subjects. PBMC from patients with guttate psoriasis responded poorly to SPEA and SPEC at concentrations of 0.1 and 1 ng/ml as compared with those from patients with plaque psoriasis, but showed high responses to SEB and TSST-1. The hyporesponsiveness recovered after improvement of the skin eruption. There was no significant difference between guttate and chronic types of psoriasis in the percentage of circulating T-cell receptor BV2 or BV8-bearing T cells, responsive to streptococcal superantigens, indicating that T-cell clonal anergy was a mechanism underlying the hyporesponsiveness. Our results suggest that superantigens released from focally infecting S. pyogenes induce a transient activation of relevant T cells, leading to the development of skin eruption and, subsequently, temporary T-cell anergy to these toxins.

  1. Phenotypic and Functional Analysis of Porcine T Lymphocytes

    Institute of Scientific and Technical Information of China (English)

    李华; 陈应华

    2001-01-01

    Porcine and other higher mammals express clusters of differentiation (CD) antigens on the surface of T lymphocytes, such as CD2, CD3, CD4, CD8, etc. However, in porcine, a high percentage of the CD4+ CD8-T lymphocyte subpopulation exist in the peripheral blood and the ratio of the CD4+ and CD8+ T lymphocyte subpopulations is reversed. These differences bring new challenges to better understanding of the phenotype and function of porcine T lymphocytes in antigen recognition and immune response.

  2. Inter-individual variability in the response of human peripheral blood lymphocytes to ionizing radiation: comparison of the dicentric and micronucleus assays.

    Science.gov (United States)

    Pajic, Jelena; Rakic, Boban; Rovcanin, Branislav; Jovicic, Dubravka; Novakovic, Ivana; Milovanovic, Aleksandar; Pajic, Vesna

    2015-08-01

    Ionizing radiation can induce a wide range of DNA damage that leads to chromosomal aberrations. Some of those aberrations (dicentrics and micronuclei) are applied in biodosimetry. Biological dosimetry assumes similar radiosensitivity of each donor, but it does not exclude inter-individual variations in radiation susceptibility. Therefore, for biological reasons, it is always challenging to investigate inter-individual variability in response to radiation. For mechanistic reasons, it is also interesting to investigate the correlation between dicentric and micronuclei formation in response to radiation. In this experiment, irradiated blood specimens from 14 healthy male and female donors have been used to evaluate inter-individual variability in response to the genotoxic effects of X-ray radiation, as well as the dose-response relationship and test sensitivity using two endpoints (dicentrics and micronuclei). The results showed similar patterns of cytogenetic biomarker distribution between donors, but differences in the response of some donors at some doses. Data also showed that responses of male donors were better detected using the dicentric test, while for females, micronucleus frequencies were higher in response to the same dose of radiation. No influence of smoking status or age on specific responses was observed. Group variability in response to radiation was evaluated using coefficient of variation for each group of individuals irradiated with the same doses; as the dose increases, group variability becomes substantially lower. Despite sporadic inter-individual variability, trend of radiation-induced changes was similar. Produced calibration curves for both types of damage revealed dicentrics as genetic damage more typical for radiation than micronuclei.

  3. File list: ALL.Bld.20.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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  5. Abnormalities of lymphocyte function and phenotypic pattern in a case of toxic epidermal necrolysis

    DEFF Research Database (Denmark)

    Hagdrup, H; Tønnesen, E; Clemmensen, O

    1992-01-01

    We examined the blood lymphocyte function and phenotypic pattern in a patient with toxic epidermal necrolysis after taking salazopyrin. We studied cell surface markers, natural killer cell activity and mitogen-induced lymphocyte transformation. Our results point to temporary immunosuppression...... as evidenced by lymphopenia with a large "null cell" population, reduced natural killer cell activity, and impaired lymphocyte response to mitogens....

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  4. File list: His.Bld.05.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  5. File list: Oth.Bld.20.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  6. File list: Unc.Bld.10.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  7. File list: Pol.Bld.10.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  8. File list: His.Bld.50.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  9. File list: Oth.Bld.05.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  10. File list: DNS.Bld.20.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  11. File list: ALL.Bld.05.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  12. File list: Pol.Bld.20.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  13. File list: Oth.Bld.10.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  14. File list: Pol.Bld.10.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  15. File list: Oth.Bld.50.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  16. File list: Oth.Bld.50.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  17. File list: DNS.Bld.10.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  18. File list: DNS.Bld.05.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  19. File list: Pol.Bld.05.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  20. File list: DNS.Bld.05.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  1. File list: Pol.Bld.50.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  2. File list: DNS.Bld.20.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  3. File list: ALL.Bld.50.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  4. File list: Pol.Bld.50.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  5. File list: Unc.Bld.10.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  6. File list: Pol.Bld.05.AllAg.T-Lymphocytes,_Regulatory [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Bld.05.AllAg.T-Lymphocytes,_Regulatory mm9 RNA polymerase Blood T-Lymphocytes, Regu...latory http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Bld.05.AllAg.T-Lymphocytes,_Regulatory.bed ...

  7. File list: Oth.Bld.05.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  8. File list: Oth.Bld.50.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  9. File list: NoD.Bld.20.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  10. File list: Oth.Bld.05.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  11. File list: ALL.Bld.05.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  12. File list: Oth.Bld.10.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  13. File list: Unc.Bld.05.AllAg.Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  14. File list: DNS.Bld.10.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  15. File list: Unc.Bld.20.AllAg.Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  16. File list: ALL.Bld.10.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  17. File list: DNS.Bld.10.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  18. File list: DNS.Bld.50.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Bld.50.AllAg.B-Lymphocytes hg19 DNase-seq Blood B-Lymphocytes ERX337094,SRX1894...09,SRX189387 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Bld.50.AllAg.B-Lymphocytes.bed ...

  19. File list: ALL.Bld.05.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  20. File list: Unc.Bld.50.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  1. File list: InP.Bld.05.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  2. File list: InP.Bld.20.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  3. File list: ALL.Bld.50.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  4. File list: ALL.Bld.05.AllAg.Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  5. File list: Pol.Bld.05.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  6. File list: His.Bld.10.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  7. File list: Oth.Bld.20.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  8. File list: Oth.Bld.10.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  9. File list: ALL.Bld.50.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  10. File list: InP.Bld.50.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  11. File list: Unc.Bld.05.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  12. File list: InP.Bld.50.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  13. File list: InP.Bld.20.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  14. File list: InP.Bld.20.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  15. File list: His.Bld.20.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  16. File list: DNS.Bld.50.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  17. File list: Pol.Bld.10.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  18. File list: ALL.Bld.05.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  19. File list: InP.Bld.50.AllAg.T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  20. File list: His.Bld.20.AllAg.B-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  1. File list: Unc.Bld.10.AllAg.T-Lymphocytes [Chip-atlas[Archive

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    Lifescience Database Archive (English)

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  3. File list: ALL.Bld.10.AllAg.T-Lymphocytes [Chip-atlas[Archive

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  6. File list: Oth.Bld.20.AllAg.B-Lymphocytes [Chip-atlas[Archive

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  8. File list: NoD.Bld.10.AllAg.B-Lymphocytes [Chip-atlas[Archive

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  9. Transfer of cholesterol from macrophages to lymphocytes in culture.

    Science.gov (United States)

    de Bittencourt Júnior, P I; Curi, R

    1998-02-01

    A major feature of macrophage metabolism is its capacity to produce and export cholesterol. Several reports have shown that the manipulation of lymphocyte cholesterol content elicits important changes in lymphocyte proliferation. These findings lead to an inquiry as to whether macrophage-derived cholesterol released into the lymphocyte surroundings may be transferred to the latter thus affecting lymphocyte function. In this study, cholesterol transfer from macrophages to lymphocytes was examined in vitro using rat cells in culture. The findings indicate that there may be a significant transfer of cholesterol from [4-14C]cholesterol labeled resident peritoneal macrophages to mesenteric lymph node resting lymphocytes (up to 173.9 +/- 2.7 pmol/10(7) lymphocytes/10(7) macrophages when co-cultivated for 48 h), in a lipoprotein-dependent manner. This represents the mass transfer of ca. 17 nmoles of cholesterol molecules per 10(7) lymphocytes from 10(7) macrophages (calculated on the basis of specific radioactivity incorporated into macrophages after the pre-labelling period), which suggests that macrophages are capable of replacing the whole lymphocyte cholesterol pool every 21 h. Moreover, an 111%-increase in the total cholesterol content of lymphocytes was found after co-cultivation with macrophages for 48 h. When compared to peritoneal cells, monocytes/macrophages obtained from circulating blood leukocytes presented a much higher cholesterol transfer capacity to lymphocytes (3.06 +/- 0.10 nmol/10(7) lymphocytes/10(7) macrophages co-cultivated for 24 h). Interestingly, inflammatory macrophages dramatically reduced their cholesterol transfer ability (by up to 91%, as compared to resident macrophages). Cholesterol transfer may involve a humoral influence, since it is not only observed when cells are co-cultivated in a single-well chamber system (cells in direct contact), but also in a two-compartment system (where cells can communicate but not by direct contact). Co

  10. The approaches in detecting cell cycle specificity of Fas-mediated apoptosis in leukemia cell lines and activated PBLs in vitro%体外Fas介导细胞凋亡的细胞周期特异性的检测方法

    Institute of Scientific and Technical Information of China (English)

    何小军; 胡静; 李小兰

    2006-01-01

    Objective: To establish a system in detecting the cell cycle specificity induced by recombinant human Fas ligand in vitro,so as to provide a reliable platform for further exploring the mechanism of cell cycle control and regulation in Fas-mediated apoptosis.Methods: The target cells-leukaemia cell lines and activated peripheral blood lymphocytes stimulated by phytohemagglutinin were incubated with recombinant human Fas ligand for 6 to 36 h,apoptosis was detected by sub-G1,common annexin-V/PI and modified annexin V and propidium iodide (API) methods and analysed by flow cytometry.Results: The modified API method demonstrated that Fas-mediated apoptosis was cell cycle specific and initiated at G1 phase.The common annexinV/PI method showed the most appropriate condition for the detection of typical cell cycle-specific apoptosis.The sub-G1 method could only illuminate late apoptosis and DNA histogram.Conclusion: Fas-mediated apoptosis was cell cycle-specific and initiated at G1 phase.Based on the modified APl and common AnnexinV/PI methods,the establishment of stable and typical cell cycle-specific model in Fas-mediated apoptosis in vitro was feasible.

  11. Prenatal ontogeny of lymphocyte subpopulations in pigs.

    Science.gov (United States)

    Sinkora, M; Sinkora, J; Reháková, Z; Splíchal, I; Yang, H; Parkhouse, R M; Trebichavsk, I

    1998-12-01

    Although porcine lymphocytes have been classified into numerous subpopulations in postnatal animals, little is known about the ontogeny of these complex cell subsets. Using double- and triple-colour flow cytometry (FCM), we investigated the surface phenotype of fetal lymphoid cells in the thymus, cord blood, spleen and mesenteric lymph nodes at different stages of gestation. It was found that the major lymphocyte subpopulations started to appear at the beginning of the second third of the gestation period, with B cells being the earliest lymphocyte subpopulation to appear in the periphery. The T-cell receptor (TCR) gamma delta+ cells were the earliest detectable T-cell subset, developing first in the thymus and subsequently arriving in the periphery. Later in ontogeny, however, the number of TCRalpha beta+ lymphocytes rapidly increased, becoming the predominant T cells both in the thymus and in the periphery. Cells with the phenotype of adult natural killer cells were also identified in pig fetuses, though their nature and functional roles remain to be investigated. In addition, CD2 was expressed on most B cells whilst very few CD4+ TCRalpha beta+ cells or CD2+ TCRgamma delta+ cells expressed CD8, suggesting that the expression of CD2 and CD8 may reflect the functional status of the cells in postnatal animals. Taken together, this study has provided a systematic analysis of fetal porcine lymphocyte subpopulations and may provide the base for studies to establish the physiological roles of these lymphocyte subsets.

  12. Bendamustine Plus Alemtuzumab for Refractory Chronic Lymphocytic Leukemia (CLL)

    Science.gov (United States)

    2013-08-20

    Recurrent Small Lymphocytic Lymphoma; Refractory Chronic Lymphocytic Leukemia; Stage III Chronic Lymphocytic Leukemia; Stage III Small Lymphocytic Lymphoma; Stage IV Chronic Lymphocytic Leukemia; Stage IV Small Lymphocytic Lymphoma

  13. Lymphocytes in patients with psoriasis promote proliferation of keratinocytes

    Institute of Scientific and Technical Information of China (English)

    DENG An-mei; ZHONG Ren-qian; CHEN Sun-xiao; ZHOU Ye; KONG Xian-tao

    2002-01-01

    Objective: To analyze the effect of lymphocytes on proliferation of keratinocytes in patients with psoriasis. Methods: Lymphocytes in lesion and peripheral blood were isolated and amplified, then cultured together with normal keratinocytes. By MTT method, the living cells were quantified in the mixed culture.Results: Compared with normal controls, lymphocytes from lesion and peripheral blood of psoriasis both promote the proliferation of keratinocytes (P<0. 01 and P<0. 05 respectively). The concentrations of IL-2 and IFN-γ in the mixture of lesion lymphocytes and keratinocytes were significantly higher than that of controls.Tripterygium glycosides inhibited this promotion. Conclusion: Lymphocytes in patients with psoriasis (mainly Thl cell) play an important role in proliferation of keratinocytes. This psoriasis cell model is useful for studies on signal transduction in psoriasis.

  14. What Is Chronic Lymphocytic Leukemia?

    Science.gov (United States)

    ... Chronic Lymphocytic Leukemia (CLL) About Chronic Lymphocytic Leukemia What Is Chronic Lymphocytic Leukemia? Cancer starts when cells ... body, including the lymph nodes, liver, and spleen. What is leukemia? Leukemia is a cancer that starts ...

  15. Suppressive effects of antigens on the activity of specific activated lymphocytes: A test to define the specificity of activated lymphocytes

    Institute of Scientific and Technical Information of China (English)

    HU Jun; PAN Sheng-jun; CAI Zhen-jie; GUAN De-lin; LIU Xiao-cheng

    2006-01-01

    Objective:With the regular mixed lymphocytes culture (MLC) to detect the allograft rejection, the reactivity of the activated lymphocytes (primed lymphocytes) of a recipient shows sometimes increase and sometimes decrease against the antigens from the donor, which is inconsistent with the clinical results. In order to establish a convenient method for testing the specificity of the activated lymphocytes in vitro, so as to know the rejection occurred or not by testing the existence of the specific activated lymphocytes against donor's HLA antigens in the recipient's peripheral blood. Methods: Anti-IL-2 neutralizing monoclonal antibody (anti-IL-2 N-mAb) and immunosuppressors were introduced in this test system in the presence of specific stimulators and activated lymphocytes. Results: When the activated lymphocytes were chosen from the one-way MLC 4 d to undergo re-stimulation by specific stimulators, the activity of activated lymphocytes in the treatment group was suppressed significantly compared with that in the control group. The result of this test method is consistent with the biopsy in the clinical diagnosis of rejection.Conclusion :It suggests that the activated lymphocytes can be inactivated by specific antigens in certain conditions. This can be a useful tool to define the specificity of the activated lymphocytes.

  16. Changed of NK cell and T-lymphocyte subsets in peripheral blood of patients with brucellosis%布鲁菌病患者外周血NK细胞和T淋巴细胞亚群的变化

    Institute of Scientific and Technical Information of China (English)

    贾宇臣; 其其格; 郭菊红; 赵海珍; 乌云; 奥敦托娅

    2014-01-01

    目的:了解布鲁菌病患者性别、年龄、职业分布特征及并发症,外周血NK细胞和T淋巴细胞亚群的变化及意义。方法对101例布鲁菌病患者采用流行病学调查法进行调查,采用流式细胞仪抗体双表法检测38例布鲁菌病和非布鲁菌病发热患者外周血NK细胞(CD16+CD56+)、淋巴细胞绝对值(CD45+)、T淋巴细胞绝对值(CD3+)、T辅助细胞绝对值(CD3+CD4+)、T抑制/细胞毒细胞绝对值(CD3+CD8+)、B淋巴细胞绝对值(CD19+)、CD4/CD8比值,并进行分析。结果布鲁菌病患者以男性居多,中老年患者居多,发病人群主要为农牧民及从事防疫工作者,并发症以肝功能损害最多;布鲁菌病患者外周血NK绝对值显著低于非布鲁菌病发热患者组,具有统计学意义(t =-2.58,P <0.05)。结论布鲁菌病以男性中老年患者居多,以农牧民和防疫员为主,布鲁菌病患者外周血NK细胞常受损伤。%Objective To investigate the characteristics of gender, age, occupation distributions and complication of patients with brucellosis, explore the changes of NK cell and T-lymphocyte subsets, and the related significance. Methods Total of 101 cases of brucellosis were analyzed by descriptive epidemiological study. NK and T/B-lymphocyte subsets were studied in peripheral blood of 38 patients with brucellosis and 35 cases with non-brucellosis fever. Results The majority of cases were male elderly patients, mainly occurred among farmers, herdsmen and stuff engaged in the epidemic prevention. The complication of brucellosis was priority to liver damage. The number of NK cell in patients with brucellosis was below compared with in patients with non-brucellosis fever, with signiifcant differeces (t=-2.58, P<0.05). Conclusions The majority of patients with brucellosis were male elderly person. Outbreak mainly occur in farmers, herdsmen and stuffs engared in epidemic prevention ifeld

  17. Identification biomarkers for cervical cancer in peripheral blood lymphocytes by oligonucleotide microarrays%应用寡核苷酸芯片筛选宫颈癌患者外周血生物标志物的研究

    Institute of Scientific and Technical Information of China (English)

    盛洁; 张为远

    2010-01-01

    Objective To identify the molecular biomarkers for cervical cancer in peripheral blood lymphocytes by oligonucleatide microarrays. Methods Human genome oligonucleotide microarray analysis included 4 early-stage cervical cancer patients and 3 controls. The selected genes from the microarray analysis were validated in additional 20 early-stage cervical cancer patients and 15 controls by real-time reverse-transcription polymerase chain reaction (RT-PCR). Results Genes identified by gene selection program were expressed differently in the blood samples of early-stage cervical cancer from those of healthy controls. To validate the gene expression data, 5 genes were analyzed by real-time RT-PCR. In three of 5 identified genes, tenasin-c, nucleolin, and enolase 2 (ENO2) showed a significant up-regulation in blood samples of early-stage cervical cancer patients versus that of the controls. Conclusion The up-regulation of tenasin-c, nucleolin and ENO2 in peripheral blood may be used to identify novel blood biomarkers for detecting cervical cancer in a clinically accessible surrogate tissue. Thus it may offer a possibility of developing a non-invasive and predictive diagnostic tool for the disease.%目的 应用人类全基因组寡核苷酸芯片技术在宫颈癌患者外周血中确定生物分子标志物.方法 从24例早期宫颈癌患者和18例正常对照者的外周血淋巴细胞中提取总RNA.应用微阵列技术,采用人类全基因组寡核苷酸芯片检测4例宫颈癌患者和3例正常对照者的差异表达基因,再对20例宫颈癌患者和15例正常对照者将初步筛选出的5个候选基因用实时定量逆转录多聚酶链反应(RT-PCR)的方法进行验证.结果 筛选得到57个差异表达基因,其中38个基因表达上调,19个基因表达下调;对初步筛选出的5个候选基因经实时定量逆转录多聚酶链反应的方法进行验证后发现黏合素-C、核仁素和磷酸丙酮酸水合酶2(enolase 2,ENO2)基因在宫

  18. Correlation between differential blood count and T-lymphocyte subset count in the early period after liver transplantation%肝移植术后早期白细胞分类计数与T淋巴细胞亚群计数的相关性研究

    Institute of Scientific and Technical Information of China (English)

    邹游; 张磊; 李瑞东; 郭闻渊

    2012-01-01

    目的 探讨肝移植术后早期白细胞分类计数与T淋巴细胞亚群计数的相关性.方法 收集84例行经典原位肝移植术受者术后第7天在不应用糖皮质激素情况下的血样本,分析T淋巴细胞亚群计数与白细胞计数、中性粒细胞计数、淋巴细胞计数及单核细胞计数的相关性.结果 Pearson分析显示,CD4+和CD8+T淋巴细胞计数与白细胞计数、中性粒细胞计数及单核细胞计数间均不相关(P值均>0.05),与淋巴细胞计数间均呈正相关(R值分别为0.778、0.651,P值均<0.001).Spearman分析显示,CD4+和CD8+T淋巴细胞计数与白细胞计数、中性粒细胞计数及单核细胞计数间均不相关(P值均>0.05),与淋巴细胞计数间均呈正相关(R值分别为0.748、0.702,P值均<0.001).非感染组的淋巴细胞计数、CD4+T淋巴细胞计数、CD8+T淋巴细胞计数均显著高于感染组(P值均<0.05).结论 肝移植术后早期淋巴细胞计数与CD4+T淋巴细胞计数具有较好的正相关性,可间接反应CD4+T淋巴细胞计数水平.%Objective To explore the correlation between differential blood count and T-lymphocyte subset count so as to provide a method to quickly identify cellular immune function in the early period after liver transplantation. Methods Blood samples were taken from 84 recipients (without application of glucocorticoid) on day 7 after orthotopic liver transplantation (OLT). The correlation between T-lymphocyte subset count and differential blood count including leucocyte count, granulocyte count and monocyte count was analyzed. Results Pearson analysis showed that the counts of CD4+ and CD8+ T lymphocytes were not correlated with the count of leucocyte, granulocyte and monocyte (all P>0.05), but were positively related to lymphocyte count (R =0.778, 0.651; both P0.05), and were positively related to lymphocyte count (R =0.748, 0.702; both P<0.001). The counts of lymphocyte, CD4+ T lymphocyte and CD8+ T lymphocyte in the

  19. 泛发性湿疹患者外周血淋巴细胞亚群的研究%Research of lymphocyte subpopulation in peripheral blood of generalized eczema patients

    Institute of Scientific and Technical Information of China (English)

    韩莹; 程欣

    2015-01-01

    Objective To investigate changes of immunologic function in generalized eczema patients. Methods Lymphocyte subpopulation in peripheral blood of 200 generalized eczema patients (generalized eczema group) were detected by flow cytometry method, and they were compared with those in 200 normal people (normal group).Results Compared with the normal group, the generalized eczema group had decreased CD3+ T cells, CD3+CD4+ T cells, CD3-CD16+56+ NK cells, CD4+/CD8+ T cells, and increased CD3+CD8+ T cells. The difference had statistical significance (P<0.05).Conclusion Immune function disorder is one of the main mechanism of generalized eczema patients, and it acts with important effect in occurrence and progress of generalized eczema.%目的:探讨泛发性湿疹患者的免疫功能变化。方法应用流式细胞仪分析检测200例泛发性湿疹患者(泛发性湿疹组)外周血中的淋巴细胞亚群,并与200例正常人(正常组)进行比较。结果与正常组相比泛发性湿疹组的CD3+ T细胞降低, CD3+CD4+ T细胞降低, CD3+CD8+ T细胞升高, CD3-CD16+56+ NK细胞降低, CD4+/CD8+降低,差异有统计学意义(P<0.05)。结论免疫功能的紊乱是泛发性湿疹患者发病的一个重要机制,在泛发性湿疹的发生、发展过程中可能起到重要的作用。

  20. Association of polymorphisms in AhR, CYP1A1, GSTM1, and GSTT1 genes with levels of DNA damage in peripheral blood lymphocytes among coke-oven workers

    Energy Technology Data Exchange (ETDEWEB)

    Yongwen Chen; Yun Bai; Jing Yuan; Weihong Chen; Jianya Sun; Hong Wang; Huashan Liang; Liang Guo; Xiaobo Yang; Hao Tan; Yougong Su; Qingyi Wei; Tangchun Wu [Huazhong University of Science and Technology, Wuhan (China). Institute of Occupational Medicine and Ministry of Education Key Lab of Environment and Health

    2006-09-15

    Accumulating evidence has shown that both DNA damage caused by the metabolites of polycyclic aromatic hydrocarbons (PAH) and genetic polymorphisms in PAH-metabolic genes contribute to individual susceptibility to PAH-induced carcinogenesis. However, the functional relevance of genetic polymorphisms in PAH-metabolic genes in exposed individuals is still unclear. In this study of 240 coke-oven workers (the exposed group) and 123 non-coke-oven workers (the control group), we genotyped for polymorphisms in the AhR, CYP1A1, GSTM1, and GSTT1 genes by PCR methods, and determined the levels of DNA damage in peripheral blood lymphocytes using the alkaline comet assay. It was found that the ln-transformed Olive tail moment (Olive TM) values in the exposed group were significantly higher than those in the control group. Furthermore, in the exposed group, the Olive TM values in subjects with the AhR Lys{sup 554} variant genotype were higher than those with the AhR Arg{sup 554}/Arg{sup 554} genotype. Similarly, the Olive TM values in the non-coke-oven workers with the CYP1A1 MspI CC + CT genotype were lower than the values of those with the CYP1A1 MspI TT genotype. However, these differences were not evident for GSTM1 and GSTT1. These results suggested that the polymorphism of AhR might modulate the effects of PAHs in the exposed group; however, the underlying molecular mechanisms by which this polymorphism may have affected the levels of PAH-induced DNA damage warrant further investigation.

  1. Immunophenotypic lymphocyte profiles in human african trypanosomiasis.

    Directory of Open Access Journals (Sweden)

    Caroline Boda

    Full Text Available Human African trypanosomiasis (HAT is a deadly vector-born disease caused by an extracellular parasite, the trypanosome. Little is known about the cellular immune responses elicited by this parasite in humans. We used multiparameter flow cytometry to characterize leukocyte immunophenotypes in the blood and cerebrospinal fluid (CSF of 33 HAT patients and 27 healthy controls identified during a screening campaign in Angola and Gabon. We evaluated the subsets and activation markers of B and T lymphocytes. Patients had a higher percentage of CD19+ B lymphocytes and activated B lymphocytes in the blood than did controls, but lacked activated CD4+ T lymphocytes (CD25+. Patients displayed no increase in the percentage of activated CD8+ T cells (HLA-DR+, CD69+ or CD25+, but memory CD8 T-cell levels (CD8+CD45RA2 were significantly lower in patients than in controls, as were effector CD8 T-cell levels (CD8+CD45RA+CD62L2. No relationship was found between these blood immunophenotypes and disease severity (stage 1 vs 2. However, CD19+ B-cell levels in the CSF increased with disease severity. The patterns of T and B cell activation in HAT patients suggest that immunomodulatory mechanisms may operate during infection. Determinations of CD19+ B-cell levels in the CSF could improve disease staging.

  2. 脓毒症患者外周血淋巴细胞PPARγ表达和活性的变化%Changes of PPARγ protein expression and activity in peripheral blood lymphocytes of patients with sepsis

    Institute of Scientific and Technical Information of China (English)

    孙湛; 黄培志; 宋振举; 童朝阳; 朱彪; 姚晨玲

    2009-01-01

    Objective To investigate the changes of PPARγ protein expression and activity of peripheral blood lymphocytes in patients with sepsis and its association with severity and prognosis of sepsis. Method Ac-cording to the guidelines to sepsis set by ACCP/SCCM consensus conference in 2003, 48 patients with sepsis ad-mitted in Emergency and Surgical ICU from December 2007 to March 2008 were enrolled in this perspective study. Sixteen healthy individuals were selected as controls. Patients with metastatic tumors, autoimmune disease, AIDS or under immunosuppressive therapy were excluded. This study was approved by the ethical committee of Zhong-shan Hospital, Fudan University. All patients were divided into mild and severe sepsis groups. Patients were also divided into survivor and non-survivor groups as per 28-day mortality. Peripheral blood lymphocytes were isolated by using Ficoll density gradient centrifugation. PPART protein expression was determined by using Westem Blot-ting. The activity of PPARγ was analyzed by using EMSA. Differences among groups were analyzed by using one-way ANOVA. Results The protein expression and activity of PPARγ were significantly increased in mild sepsis patients (0.56±0.12 and 4.13±0.22, respectively) compared with both healthy controls (0.39±0.07 and 2.42±0.17, respectively) and severe sepsis patients (0.30±0.07 and 1.63±0.12, respectively) (P < 0.05). However, the protein expression and activity of PPARγ were obviously decreased in severe sepsis patients compared with healthy individuals and mild spsis patients (P < 0.05). Survivors from sepsis had significantly higher protein expression and activity (0.54±0.11 and 3.59±0.34, respectively) than non-survivors (0.21±0.08 and 1.94 ±0.25, respectively) (P < 0.05). Conclusions These data suggest that the protein expression and activity of PPARγ in peripheral blood lymphocytes might be valuable biomarkers in assessing the severity and outcome of pa-tients with sepsis.%目的

  3. Mean dose to lymphocytes during radiotherapy treatments

    Energy Technology Data Exchange (ETDEWEB)

    Brandan, M.E.; Perez-Pastenes, M.A. [Instituto de Fisica (Mexico); Ostrosky-Wegman, P.; Gonsebatt, M.E. [Instituto de Investigaciones Biomedicas (Mexico); Diaz-Perches, R. [Hospital General de Mexico (Mexico)

    1994-10-01

    Using a probabilistic model with parameters from four radiotherapy protocols used in Mexican hospitals for the treatment of cervical cancer, the authors have calculated the distribution of dose to cells in peripheral blood of patients. Values of the mean dose to the lymphocytes during and after a {sup 60}Co treatment are compared to estimates from an in vivo chromosome aberration study performed on five patients. Calculations indicate that the mean dose to the circulating blood is about 2% of the tumor dose, while the mean dose to recirculating lymphocytes may reach up to 7% of the tumor dose. Differences up to a factor of two in the dose to the blood are predicted for different protocols delivering equal tumor doses. The data suggest mean doses higher than the predictions of the model. 10 refs., 3 figs., 2 tabs.

  4. Role of senescence marker p16 INK4a measured in peripheral blood T-lymphocytes in predicting length of hospital stay after coronary artery bypass surgery in older adults.

    Science.gov (United States)

    Pustavoitau, Aliaksei; Barodka, Viachaslau; Sharpless, Norman E; Torrice, Chad; Nyhan, Daniel; Berkowitz, Dan E; Shah, Ashish S; Bandeen Roche, Karen J; Walston, Jeremy D

    2016-02-01

    Adults older than 65 years undergo more than 120,000 coronary artery bypass (CAB) procedures each year in the United States. Chronological age alone, though commonly used in prediction models of outcomes after CAB, does not alone reflect variability in aging process; thus, the risk of complications in older adults. We performed a prospective study to evaluate a relationship between senescence marker p16(INK4a) expression in peripheral blood T-lymphocytes (p16 levels in PBTLs) with aging and with perioperative outcomes in older CAB patients. We included 55 patients age 55 and older, who underwent CAB in Johns Hopkins Hospital between September 1st, 2010 and March 25th, 2013. Demographic, clinical and laboratory data following outline of the Society of Thoracic Surgeons data collection form was collected, and p16 mRNA levels in PBTLs were measured using TaqMan® qRT-PCR. Associations between p16 mRNA levels in PBTLs with length of hospital stay, frailty status, p16 protein levels in the aortic and left internal mammary artery tissue, cerebral oxygen saturation, and augmentation index as a measure of vascular stiffness were measured using regression analyses. Length of hospital stay was the primary outcome of interest, and major organ morbidity, mortality, and discharge to a skilled nursing facility were secondary outcomes. In secondary analysis, we evaluated associations between p16 mRNA levels in PBTLs and interleukin-6 levels using regression analyses. Median age of enrolled patients was 63.5 years (range 56-81 years), they were predominantly male (74.55%), of Caucasian descent (85.45%). Median log2(p16 levels in PBTLs) were 4.71 (range 1.10-6.82). P16 levels in PBTLs were significantly associated with chronological age (mean difference 0.06 for each year increase in age, 95% CI 0.01-0.11) and interleukin 6 levels (mean difference 0.09 for each pg/ml increase in IL-6 levels, 95% CI 0.01-0.18). There were no significant associations with frailty status, augmentation

  5. Alzheimer病患者外周血淋巴细胞Akt及其活化水平的改变及意义%Change and its significance of level of Akt and its activation in peripheral blood lymphocytes in patients with Alzheimer disease

    Institute of Scientific and Technical Information of China (English)

    左宝旭; 王珊; 生晓娜; 张国华; 赵辉; 苗江永

    2013-01-01

    目的 研究Alzheimer病(AD)患者外周血淋巴细胞Akt及其活化水平的改变及意义.方法 对20例AD患者(AD组)进行简易精神状态检查(MMSE)量表评分,用免疫印迹法检测其外周血淋巴细胞的Akt及磷酸化Akt (p-Akt)水平,并与帕金森病(PD)患者(PD组)和正常对照者(正常对照组)进行比较.分析AD患者外周血淋巴细胞的Akt水平与病程和MMSE量表评分的关系.结果 与正常对照组相比,AD组外周血淋巴细胞中Akt及p-Akt水平明显升高(均P<0.01);PD组p-Akt水平显著增高(P<0.05).与PD组相比,AD组Akt水平升高明显(P<0.05).AD患者外周血淋巴细胞中Akt水平与病程呈正相关(r=0.602,P=0.014),与MMSE量表评分呈负相关(r=-0.560,P=0.024).结论 AD患者外周血淋巴细胞的Akt及其活化水平显著增高,并且病程越长、病情越重者增高的越明显.提示Akt有可能成为AD诊断及病情评价的外周标志物.%Objective To investigate the change and its significance of level of Akt and its activation in peripheral blood lymphocytes in patients with Alzheimer disease (AD). Methods Twenty AD patients (AD group) were taken into mini mental state examination ( MMSE) tests. Their Akt and phosphated Akt ( p-Akt) levels of peripheral blood lymphocytes were detected by western blotting, and then the results were compared with that of Parkinson disease ( PD) patients (PD group) and normal controls ( NC group). The relationship among the level of Akt, the disease duration and MMSE score were analyzed. Results Compared with NC group, the levels of Akt and p-Akt in peripheral blood lymphocytes were significantly increased in AD group ( all P < 0. 01) ; and the level of p-Akt in peripheral blood lymphocytes in PD group was significantly increased (P < 0. 05). Compared with PD group, the level of Akt in peripheral blood lymphocytes in AD group was significantly increased ( P < 0. 05 ) . The level of Akt in peripheral blood lymphocytes in AD patients were

  6. 苯对外周血人淋巴细胞周期阻滞及凋亡影响%Effects of benzene on cell cycle and apoptosis of peripheral blood lymphocytes in vitro

    Institute of Scientific and Technical Information of China (English)

    段鹏; 胡春卉; 刘颖; 杨益萍; 仇小强; 韦小敏

    2011-01-01

    目的 研究苯及其代谢产物氢醌对外周血人淋巴细胞周期阻滞与凋亡的影响,探讨苯的细胞毒性作用机制.方法 离体培养人淋巴细胞24h后加S9液,设置苯低、中、高浓度(0.25、3.5、50 μmol/L)和氢醌低、中、高浓度(50、150、450 μmol/L)的染毒组,另设空白对照组和溶剂对照组,采用四甲基偶氮唑蓝比色法检测细胞相对存活率,流式细胞术检测细胞周期和凋亡的分布状况,荧光检测细胞存活率的含量,单细胞凝胶电泳技术检测DNA断裂.结果 苯与氢醌剂量依赖性降低人淋巴细胞存活率,诱导人淋巴细胞阻滞于S+ G2/M期,并明显促凋亡且随着染毒浓度升高细胞内活性氧含量增加,与对照组比较,差异有统计学意义(P<0.05),苯与氢醌高浓度组彗星尾长分别为(26.45±7.96)、(30.28 ±6.07)μm,均明显高于对照组(P<0.01).结论 苯及其代谢物氢醌在体外可导致人淋巴细胞存活率降低,细胞周期紊乱,其机制与细胞内活性氧产生及DNA -蛋白质损伤有关.%Objective To study the effects of benzene and the benzene metabolite hydroquinone on the cell cycle and apoptosis of peripheral blood lymphocytes, and to explore the molecular mechanisms underlying benzene-induced cytoxicity damage. Methods Human lymphatic cells were isolated, cultivated and then divided into three groups including of low, moderate,and high benzene exposure(0. 25,3. 5,50 μmol/L) and three groups of low,moderate,and high hydroquinone exposure(50,150,450 μmol/L). One solvent control and one blank control group were also set up. After the treatment,we assayed the growth arrest of lymphocytes induced by benzene and hydroquinone by methyl thiazolyl tetrazolium( MTT) test. Row cytometry was applied to detect cell cycle and apoptosis rate. 2,7-dichlorodihydro fluorescein diacetate( DCFH-DA) assay was used to detect reactive oxygen species(ROS) contents. Lymphocytes'DNA fragment was detected with single cell

  7. 卷烟烟气致人淋巴细胞细胞毒性和遗传毒性的体外试验%Cyto-genotoxicity induced by cigarette smoke condensates in human peripheral blood lymphocytes in vitro

    Institute of Scientific and Technical Information of China (English)

    楼建林; 何继亮; 周国俊; 储国海; 黄芳芳; 蒋健; 郑树; 陆叶珍; 李晓雪; 陈志健

    2009-01-01

    Objective To investigate the cyto-genotoxicity of cigarette smoke condensates (CSCs) in human peripheral blood lymphocytes with different assays in vitro. Methods Human lymphocytes were ex-posed to particle matter of cigarette smoke combined with or without S9 mixtures at doses of 25,50,75,100 and 125 ug/ml for 3 h. The cytotoxicity induced by CSCs was detected by CCK-8 assay. The DNA damage, DNA repair (repair time: 30, 60, 90, 120 and 240 min, respectively) and the somatic cell mutations induced by 75 ug/ml CSCs were measured by comet assay, hprt gene and TCR gene mutation tests, respectively. Results CCK-8 assay indicated that the cell viability decreased with CSCs doses. At the doses of 100,125 ug/ml, the cell viability of CSCs +S9 group was significantly higher than that of CSCs -S9 group(P<0.05,P< 0.01). In comet assay, DNA damage significantly increased in a dose-dependent manner, as compared with controls (P<0.01). Moreover, there was significant difference between -S9 group and +S9 group (P<0.05, P< 0.01). The Mf-TCR at each dose group was significantly higher than that of controls (P<0.05 ,P<0.01). The Mf-hprt at high-dose groups were significantly higher than that of controls (P<0.01), and significant difference of Mf-TCR and Mf-hprt at high doses of CSCs between -S9 group and +S9 group (P<0.05,P<0.01). The DNA damage induced by CSCs+S9 or CSCs-S9 could be repaired, but DNA repair speed was different between -S9 group and +S9 group (P<0.05, P<0.01). Conclusion CSCs may induce cyto-genotoxicity in human periph-eral blood lymphocytes in vitro, but S9 mix could reduce the toxicity of CSCs and impact DNA repair speed.%目的 用不同体外试验评价卷烟烟气粒相物提取液(CSCs)致人外周血淋巴细胞的细胞毒性和遗传毒性.方法 分别以25、50、75、100和125 ug/ml的CSCs在加或不加肝脏微粒体酶(S9)系统下作用于人外周血淋巴细胞3 h,然后用CCK-8试验检测细胞毒性,用彗星试验检测DNA损伤,用hprt

  8. 空调病患者外周血细胞及T淋巴细胞亚群检测的意义%Significance of changes of peripheral blood cells and T-lymphocyte subgroups in patients with aircondition disease

    Institute of Scientific and Technical Information of China (English)

    孙成栋; 李真; 文艳

    2011-01-01

    目的 探讨空调病患者外周血细胞及淋巴细胞亚群变化的特点及意义.方法 用血常规及流式细胞术分析36例发热空调病患者(发热组)及30例非发热患者(非发热组)外周血细胞及淋巴细胞亚群的变化,并以30例健康人(健康组)作为对照进行分析.结果(1)3组患者白细胞比较无明显差别(F=1.89,P=0.134);(2)发热组、非发热组、健康组中性粒细胞百分比分别为(68.13±5.67)%、(53.24±8.36)%、(61.25±6.31)%,发热组与非发热组、健康组比较中性粒百分比明显升高(t=6.64,9.13;P=0.018,0.027),非发热组与健康组比较无明显改变(P>0.05);淋巴细胞百分比为(31.66±7.11)%、(43.67±8.17)%、(30.98±8.76)%,与发热组、健康组比较非发热组血淋巴细胞百分比明显升高(t=5.59,4.98;P=0.022,0.021);(3)CD4+百分比分别为(36.351±6.531)%、(42.676±5.169)%、(39.253±5.461)%,与非发热组比较发热组明显下降(t=1.28,P=0.039),与健康组比较非发热组CD4+细胞明显上升(t=3.80,P=0.048);CD8+百分比分别为(28.084±7.344)%、(21.186±6.331)%、(24.607±7.108)%,发热组较非发热组CD8+则明显升高(t=3.17,P=0.016);(4)CD4+/CD8+百分比分别为(1.629±0.554)%、(2.380±0.391)%、(2.079±0.659)%,与非发热组及健康组比较发热组明显下降(t=19.21,6.98;P<0.001,0.006),非发热组较健康组则明显升高(t=8.68,P=0.031).结论 空调病患者淋巴细胞亚群及比率的变化提示空调病患者细胞免疫功能网络的失衡.%Objective To discuss the metabolic characteristics and significance of peripheral blood cell and T lymphocyte subgroup in patients with air condition disease.Methods Thirty-six patients diagnosed as air condition diseases with fever and 30 patients diagnosed as non-fever patients enrolled this study as observation group(fever group and non-fever group respectively),and 30 health population were used as control.The changes of peripheral blood cell and T

  9. 阿奇霉素对支气管哮喘患儿外周血辅助性T淋巴细胞9和辅助性T淋巴细胞17功能的影响%Influence of Azithromycin on peripheral blood helper T lymphocyte 9 cells and helper T lymphocyte 17 cells in ;children with bronchial asthma

    Institute of Scientific and Technical Information of China (English)

    刘伟; 周娟; 孙莹; 张秋业

    2015-01-01

    Objective To observe the influence of Azithromycin on helper T lymphocyte 9 cells ( Th9 ) and Th17 in peripheral blood of children with bronchial asthma,and to investigate the immunomodulating effect of Azithro-mycin. Methods Twenty-six asthmatic children were selected as the experimental group,and 17 healthy children as a control group. Peripheral blood mononuclear cells(PBMC)were isolated from venous blood by density gradient cen-tri-fugation under aseptic conditions. PBMC were split by phytohemagglutinin( PHA) in vitro. Different concentrations of Azithromycin (0,0. 1,1. 0,10. 0 mg/L)were added into the cultures in the experimental group. The control group was not interfered with azithromycin. The supematant was collected after 72 h. The levels of interleukin ( IL)-4, IL-9,IL-17 and IL-23 in the supematant were determined by enzyme-linked immunosorbent assay(ELISA). SPSS 17. 0 software was used to analyze data. Results (1) The levels of IL-4,IL-9,IL-17 and IL-23 produced from PBMC of the experimental group were significantly higher than those in the control group(t=9. 210,3. 040,8. 965,2. 796,P0. 05). The levels of IL-17 and IL-23 of Azithromycin 10 mg/L were lower than those in Azithromycin 0 mg/L and 0. 1 mg/L(P0. 05). (3)IL-9 level was negatively correlated with IL-4 level in the experimental group(r=-0. 255,P>0. 05). The levels of IL-23 and IL-17 secreted by Th17 in asthmatic chil-dren had correlation. The secretion of IL-17 levels rose with the secretion of IL-23 rise(r=0. 64,P0.05)。阿奇霉素10.0 mg/L 组 IL-17、IL-23的分泌水平低于0 mg/L 组和0.1 mg/L 组(P0.05);3.哮喘组患儿PBMC经不同浓度阿奇霉素干预后分泌IL-9与IL-4水平间无相关性(r=-0.255,P>0.05),哮喘组PBMC分泌IL-17与IL-23水平呈正相关(r=0.64,P<0.05)。结论支气管哮喘患儿Th9和Th17细胞功能增强;阿奇霉素在较高组织浓度(10.0 mg/L)条件下可抑制Th9和 Th17细胞功能,前者与IL-4水平变化无关联,后者与IL-23水平降

  10. Acute Lymphocytic Leukemia

    Science.gov (United States)

    ... for information in your local library and on the Internet. Good sources include the National Cancer Institute, the ... mayoclinic.org/diseases-conditions/acute-lymphocytic-leukemia/basics/definition/CON-20042915 . Mayo Clinic Footer Legal Conditions and ...

  11. Chronic Lymphocytic Leukemia

    Science.gov (United States)

    Leukemia is cancer of the white blood cells. White blood cells help your body fight infection. Your blood cells form in your bone marrow. In leukemia, the bone marrow produces abnormal white blood cells. ...

  12. Acute Lymphocytic Leukemia

    Science.gov (United States)

    Leukemia is cancer of the white blood cells. White blood cells help your body fight infection. Your blood cells form in your bone marrow. In leukemia, however, the bone marrow produces abnormal white blood ...

  13. Lymphocyte subpopulations in Sheehan's syndrome.

    Science.gov (United States)

    Atmaca, Hulusi; Araslı, Mehmet; Yazıcı, Zihni Acar; Armutçu, Ferah; Tekin, Ishak Özel

    2013-06-01

    The role of autoimmunity in the development of Sheehan's syndrome is obscure. There are a limited number of studies investigating the immunological alterations accompanying Sheehan's Syndrome. Our objective was to evaluate lymphocyte subsets in these patients. We conducted a cross-sectional clinical study. Cytofluorometry was used for the immunophenotyping of peripheral blood leukocytes from patients with Sheehan's syndrome followed up in the endocrine clinic during 2005-2009. Fifteen consecutive patients (mean age 61.6 ± 11.3, range 34-75 years) and 25 healthy controls (mean age 56.7 ± 10.6, range 34-80 years) were included. There was no statistically significant difference between the groups in terms of mean age. The percentages of CD19(+), CD16(+)/56(+), CD8(+)28(-), γδTCR(+), CD8(+); the total lymphocyte counts; and the ratio of CD8(+)28(-)/CD8(+)28(+) were similar (p > 0.05) between patients and controls. Whereas the leucocyte counts (p = 0.003), the percentage of CD3 (+) DR (+) (p Sheehan's syndrome compared to healthy controls. There was a positive correlation between the duration of illness and the percentage of CD3(+)DR(+) (r = 0.53, p = 0.03) expression. Some peripheral lymphocyte cell subsets show marked variation in patients with Sheehan's syndrome in comparison to matched healthy subjects, which may have implications for altered immune regulation in these patients. High CD3 (+) DR (+) expression that correlates with the duration of illness in Sheehan's patients is suggestive of an ongoing inflammation accompanying the slow progression of pituitary dysfunction in Sheehan's syndrome. It is not clear if these cellular alterations contribute to the cause or consequence of pituitary deficiency in Sheehan's syndrome.

  14. X-ray irradiation affects Gadd45 mRNA expression in human peripheral blood lymphocytes%X 射线对人周围血淋巴细胞 Gadd45 mRNA 表达影响

    Institute of Scientific and Technical Information of China (English)

    王恰; 李明芳; 杨爱初; 杨宇华; 梁晓阳; 严茂胜

    2015-01-01

    Objective To analyze the relative expression level of Growth arrest and DNA damage gene (Gadd)45 mRNA of human peripheral blood lymphocytes after X-ray irradiation with different doses and time,and to explore the possibility of Gadd45 gene as a biological dosimeter.Methods i)Dose-effect experiment.The healthy human peripheral blood were irradiated by 0.00,0.10,0.25,0.50,1.00,2.00,3.00 and 5.00 Gy X-ray and the lymphocytes were separated for measurement.ii)Time-effect experiment.The healthy human peripheral blood were irradiated by 2.00 Gy X-ray and the lymphocytes were separated and then cultivate for measurement after cultivation time of 0,3,6,12,24 and 48 hours.The real-time fluorescent quantitative polymerase chain reaction method was used to detect Gadd45 mRNA expression level. Results i)The relative expression level of Gadd45 mRNA increased with the increasing dose from 0.00 to 0.50 Gy.The linear regression equation was ^y =1.056 9 +7.132 2 x (determination coefficient was 0.992,P <0.01 ).The relative expression level of Gadd45 mRNA achieved the peak value when the irradiation dose was 0.50 Gy,which was 4.53 times of that with 0.00 Gy dose irradiation.The relative expression level of Gadd45 mRNA gradually decreased in 0.50-5.00 Gy.ii)In the period of 3-12 hours after 2.00 Gy of X-ray irradiation,the relative expression level of Gadd45 mRNA increased with the increasing exposure time.The linear regression equation was ^y =-6.366 0 +4.965 0 x (determination coefficient was 0.932,P <0.05).It achieved the peak value at the time of 12 hours,which was 57.68 times of that with 0-hour time group.During the time of 24-48 hours after irradiation,the relative expression level of Gadd45 mRNA decreased sharply,the 48-hour group was 2.08 times of 0-hour time group.Conclusion X-ray irradiation up-regulated the expression level of Gadd45 mRNA.There was a good dose-effect and time-effect relation in specific dose and time range.Further study is needed to explore if Gadd45

  15. Analysis on chromosomal aberrations of peripheral blood lymphocytes in radiological workers in Shenzhen City%深圳市放射工作人员周围血淋巴细胞染色体畸变分析

    Institute of Scientific and Technical Information of China (English)

    刘征宇; 杨学琴; 惠长野; 高朝贤; 李丽梅; 陈郁筠; 曹建伟

    2013-01-01

    [Objective] To analyze the occupational health damage in radiological worker in Shenzhen City,provide the data for assessing the effect of radiation protection.[Methods] 825 radiological workers received the radiation hygienic monitoring from 2008-2011 in Shenzhen City were chosen as the objectives,and the chromosomal aberrations of peripheral blood lymphocytes were detected.The types of chromosomal aberrations were compared between different year,different working year,different gender,and different work type of workers.[Results] During 2008-2011,the annual rates of chromosomal aberration in radiological workers were significantly higher than that in the control group (all P < 0.05).The annual rate of chromosomal aberration from high to low was in 2008,2009,2011and 2010.There was no significant difference between males and females (P > 0.05).The rates of chromosomal aberrations increased with radiation exposure age,and the highest rate was observed in workers with more than 30 years of service,which was significantly higher than groups (P < 0.05).The rate of chromosomal aberrations in industrial radiological workers was significantly higher than that in medical radiological workers,and the rate in both two groups were significantly higher than that in the control group (P < 0.05).The incidence rate of acentric aberrations (acentric fragment,minute and acentric ring)was significantly higher than that of dicentric (dic) and ring (r) chromosomes.[Conclusion] Long-term exposure to low dose of ionizing radiation may cause the increasing chromosomal aberrations of peripheral blood lymphocyte in radiological workers,so it is necessary to strengthen the radiation protection to reduce the ionizing radiation injury.%目的 分析深圳市放射工作人员的职业健康损害情况,为评价放射卫生防护工作提供依据.方法 对该市2008-2011年4年间接受放射卫生监护健康检查的825名放射工作人员进行周围血淋巴细胞染色体畸

  16. Genotoxicity of the herbicide butachlor in cultured human lymphocytes.

    Science.gov (United States)

    Sinha, S; Panneerselvam, N; Shanmugam, G

    1995-08-01

    Butachlor, a pre-emergence herbicide was investigated for its ability to induce sister chromatid exchanges (SCE) and chromosome aberrations (CA) in cultured human peripheral blood lymphocytes. Mitogen-stimulated lymphocytes were treated with three different concentrations (5, 10 and 20 micrograms/ml) of butachlor for 24, 48 and 72 h. Our results indicate a dose-dependent increase in the frequency of chromosomal aberrations at 24, 48 and 72 h of treatment with butachlor. No SCE was promoted by butachlor.

  17. Evaluation of lymphocyte subgroups in children with subacute sclerosing panencephalitis.

    Science.gov (United States)

    Yilmaz, C; Yuca, S A; Yilmaz, N; Oner, A F; Caksen, H

    2009-01-01

    The aetiology of subacute sclerosing panencephalitis (SSPE) remains to be fully elucidated, although it follows infection with a hypermutant defective M-protein measles virus. This study analysed peripheral blood lymphocyte subgroups to determine their role in the pathophysiology of SSPE. It included 22 children with SSPE aged 2 - 15 years (patient group) and 22 age- and gender-matched healthy children (control group). In children or= 6 years old, there were no significant differences in the lymphocyte subgroups. In conclusion, these findings suggest that a low CD4(+) lymphocyte count might be responsible for SSPE in younger children.

  18. File list: NoD.Bld.05.AllAg.CD8-Positive_T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  1. File list: Pol.Bld.50.AllAg.CD8-Positive_T-Lymphocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  10. File list: Unc.Bld.10.AllAg.CD8-Positive_T-Lymphocytes [Chip-atlas[Archive

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  11. Endothelial PI 3-kinase activity regulates lymphocyte diapedesis.

    Science.gov (United States)

    Nakhaei-Nejad, Maryam; Hussain, Amer M; Zhang, Qiu-Xia; Murray, Allan G

    2007-12-01

    Lymphocyte recruitment to sites of inflammation involves a bidirectional series of cues between the endothelial cell (EC) and the leukocyte that culminate in lymphocyte migration into the tissue. Remodeling of the EC F-actin cytoskeleton has been observed after leukocyte adhesion, but the signals to the EC remain poorly defined. We studied the dependence of peripheral blood lymphocyte transendothelial migration (TEM) through an EC monolayer in vitro on EC phosphatidylinositol 3-kinase (PI 3-kinase) activity. Lymphocytes were perfused over cytokine-activated EC using a parallel-plate laminar flow chamber. Inhibition of EC PI 3-kinase activity using LY-294002 or wortmannin decreased lymphocyte TEM (48 +/- 6 or 34 +/- 7%, respectively, vs. control; mean +/- SE; P structure" after intercellular adhesion molecule-1 ligation, whereas this was inhibited by jasplakinolide treatment. A similar fraction of lymphocytes migrated on control or LY-294002-treated EC and localized to interendothelial junctions. However, lymphocytes failed to extend processes below the level of vascular endothelial (VE)-cadherin on LY-294002-treated EC. Together these observations indicate that EC PI 3-kinase activity and F-actin remodeling are required during lymphocyte diapedesis and identify a PI 3-kinase-dependent step following initial separation of the VE-cadherin barrier.

  12. Chemokines, lymphocytes, and HIV

    Directory of Open Access Journals (Sweden)

    Farber J.M.

    1998-01-01

    Full Text Available Chemokines are members of a family of more than 30 human cytokines whose best-described activities are as chemotactic factors for leukocytes and that are presumed to be important in leukocyte recruitment and trafficking. While many chemokines can act on lymphocytes, the roles of chemokines and their receptors in lymphocyte biology are poorly understood. The recent discoveries that chemokines can suppress infection by HIV-1 and that chemokine receptors serve, along with CD4, as obligate co-receptors for HIV-1 entry have lent urgency to studies on the relationships between chemokines and lymphocytes. My laboratory has characterized Mig and Crg-2/IP-10, chemokines that are induced by IFN-g and that specifically target lymphocytes, particularly activated T cells. We have demonstrated that the genes for these chemokines are widely expressed during experimental infections in mice with protozoan and viral pathogens, but that the patterns of mig and crg-2 expression differed, suggesting non-redundant roles in vivo. Our related studies to identify new chemokine receptors from activated lymphocytes resulted in the cloning of STRL22 and STRL33. We and others have shown that STRL22 is a receptor for the CC chemokine MIP-3a, and STRL22 has been re-named CCR6. Although STRL33 remains an orphan receptor, we have shown that it can function as a co-receptor for HIV-1 envelope glycoproteins, and that it is active with a broader range of HIV-1 envelope glycoproteins than the major co-receptors described to date. The ability of STRL33 to function with a wide variety of envelope glycoproteins may become particularly important if therapies are instituted to block other specific co-receptors. We presume that investigations into the roles of chemokines and their receptors in lymphocyte biology will provide information important for understanding the pathogenesis of AIDS and for manipulating immune and inflammatory responses for clinical benefit

  13. Delayed hypersensitivity in the guinea-pig to a protein—hapten conjugate and its relationship to in vitro transformation of lymph node, spleen, thymus and peripheral blood lymphocytes

    Science.gov (United States)

    Oppenheim, J. J.; Wolstencroft, R. A.; Gell, P. G. H.

    1967-01-01

    Guinea-pig delayed hypersensitivity to purified protein derivative (PPD) and guinea-pig albumin-orthanilic acid (AO) was produced in the absence of detectable antibody formation to the conjugate. Ten days after sensitization the guinea-pig peripheral leucocytes, lymph nodes, spleen and thymus cell suspensions were cultured from 1 to 5 days with various concentrations of immunizing antigens, unconjugated hapten, a hapten—ovalbumin conjugate or phytohaemagglutinin (PHA). All the cultures of `draining' lymph node cells, and about 40 per cent of the spleen and peripheral leucocyte cultures manifested increased lymphocyte transformation on radioautographs, and by total tritiated thymidine incorporation when stimulated by PPD or AO. In addition all the cultures responded well to PHA. However, lymphocytes from the mediastinal and cervical lymph nodes from the immunized, and most of the lymphoid organ cultures from unimmunized guinea-pigs were not stimulated by antigens but responded only to PHA. Cultured guinea-pig thymocytes did not respond to any stimulus. The in vitro lymphocyte proliferation was carrier specific. It did not occur in response to unconjugated hapten. However, the response to AO was partially inhibited in the presence of the hapten. The in vitro kinetics and morphological changes in the cultures also were investigated, and the immunological significance and specificity of lymphocyte transformation are discussed. PMID:6017286

  14. At High Levels, Constitutively Activated STAT3 Induces Apoptosis of Chronic Lymphocytic Leukemia Cells.

    Science.gov (United States)

    Rozovski, Uri; Harris, David M; Li, Ping; Liu, Zhiming; Wu, Ji Yuan; Grgurevic, Srdana; Faderl, Stefan; Ferrajoli, Alessandra; Wierda, William G; Martinez, Matthew; Verstovsek, Srdan; Keating, Michael J; Estrov, Zeev

    2016-05-15

    In chronic lymphocytic leukemia (CLL), the increment in PBLs is slower than the expected increment calculated from the cells' proliferation rate, suggesting that cellular proliferation and apoptosis are concurrent. Exploring this phenomenon, we found overexpression of caspase-3, higher cleaved poly (ADP-ribose) polymerase levels (p < 0.007), and a higher apoptosis rate in cells from patients with high counts compared with cells from patients with low counts. Although we previously found that STAT3 protects CLL cells from apoptosis, STAT3 levels were significantly higher in cells from patients with high counts than in cells from patients with low counts. Furthermore, overexpression of STAT3 did not protect the cells. Rather, it upregulated caspase-3 and induced apoptosis. Remarkably, putative STAT3 binding sites were identified in the caspase-3 promoter, and a luciferase assay, chromatin immunoprecipitation, and an EMSA revealed that STAT3 activated caspase-3 However, caspase-3 levels increased only when STAT3 levels were sufficiently high. Using chromatin immunoprecipitation and EMSA, we found that STAT3 binds with low affinity to the caspase-3 promoter, suggesting that at high levels, STAT3 activates proapoptotic mechanisms and induces apoptosis in CLL cells.

  15. The expression of p53 gene in peripheral blood lymphocyte of acute Kawasaki disease%p53基因与川崎病患者淋巴细胞凋亡关系探讨

    Institute of Scientific and Technical Information of China (English)

    易岂建; 杨锡强; 李成荣; 张远维; 王莉佳

    2001-01-01

    目的:进一步探讨川崎病(KD)急性期患者外周血淋巴细胞凋亡延迟的机理。方法:采用斑点杂交(Dot-blot)检测淋巴细胞p53基因mRNA表达水平;流式细胞仪(FCM)检测p53蛋白质表达阳性细胞百分率。结果:KD患者外周血淋巴细胞p53基因mRNA和p53蛋白质表达水平降低,与正常儿童比较差异显著(P<0.005);当给予静脉注射免疫球蛋白(IVIG)治疗后或加入抗IL-6单抗培养时,外周血淋巴细胞p53基因mRNA和P53蛋白质表达水平提高。结论:KD急性期患者外周血淋巴细胞p53基因表达水平降低,其原因可能与本病患者异常升高的IL-6有关。p53基因具有促进细胞凋亡的作用,KD患者外周血淋巴细胞凋亡延迟可能与高浓度IL-6抑制p53基因的表达有关。%Objective: To further explore the mechanism of inhibited apoptosis of peripheral blood mononuclear cell (PBMC) in acute Kawasaki disease(KD). Methods: The expression level of p53 gene mRNA was determined by dot-blot; p53 protein positive cell percentage was detected by flow cytometry (FCM). Results:The expression of p53 gene mRNA and p53 protein in acute KD patients were decreased(P<0.001), but increased after treating with intravenous immunoglobulin(IVIG) in vivo or adding anti-IL-6 monoantibody(mAb)into PBMC culture in vitro. Conclusion. The decreased expression of p53 gene mRNA and p53 protein may be associated with the high concentration of IL-6 in KD patients, p53 gene expression could induce lymphocyte apoptosis. Thus, the expression of p53 gene inhibited by the increased IL-6 production might be related to delaying or depressing apoptosis of PBMC in KD.

  16. 肺结核合并2型糖尿病患者的T淋巴细胞亚群及其与血糖控制水平相关性研究%Study on the Correlation of T Lymphocyte Subsets and Blood Glucose Level in Patients With Pulmonary Tuberculosis Complicated With Type 2 Diabetes Mellitus

    Institute of Scientific and Technical Information of China (English)

    张砚; 朱凌云; 左维泽

    2015-01-01

    Objective To discuss the correlation of T lymphocyte subsets and blood glucose level in patients with pulmonary tuberculosis complicated with type 2 diabetes melitus. Methods Flow cytometry was used to detect the levels of peripheral blood lymphocyte subsets, and the level of glycosylated hemoglobin was detected by using the immune assay.ResultsThere had no statistical significance in CD3+, CD4+, CD8+, CD4+/CD8+ between DM+LTB and DM group(P> 0.05)( HbAlc 7%, there had statistical significance(P0.05);HbAlc >7%,DM+LTB与DM两组的CD3+,CD4+、CD8+、CD4+/CD8+有统计学差异(P<0.05)。结论长期高血糖状态会加重肺结核合并2型糖尿病患者细胞免疫功能紊乱。

  17. IDENTIFICATION OF THE GENETIC SEX OF TONGUEFISH (CYNOGLOSSUS SEMILAEVIS)BY THE METHOD OF PERIPHERAL BLOOD LYMPHOCYTIC CELL CULTIVATION AND CHROMOSOME PREPARATION%半滑舌鳎血淋巴细胞体外培养及其染色体制备在性别鉴定中的应用

    Institute of Scientific and Technical Information of China (English)

    张博; 王贤丽; 杨长庚; 刘珊珊; 胡乔木; 陈松林

    2011-01-01

    研究建立了半滑舌鳎(Cynoglossus semilaevis Giinther)外周血淋巴细胞体外培养及染色体制备方法,确定了最佳条件为:在24℃的环境中半滑舌鳎血淋巴细胞在添加20%胎牛血清的MEM培养基中,用终浓度为0.3 mg/mL的LPS为刺激源培养72h,在结束培养前3h加入终浓度0.08μg/mL秋水仙素,可获得较多、较好的染色体分裂相.利用这种方法对半滑舌鳎遗传性别进行了鉴定,同时与生理解剖观察、性腺切片、性腺细胞培养、雌性特异标记等方法进行了比较,确定了适宜各阶段不同类型鱼的性别鉴定方法,丰富了半滑舌鳎活体遗传性别鉴定的方法.%The method of the peripheral blood lym-phocytic cell culture and chromosome preparation of the tonguefish (Cynoglossus semilaevis Günther) was established in this research. The conditions of the peripheral blood lymphocytic cell culture and chromosome preparation were well searched from culturing temperature and time, effect of PHA, ConA,Lipopolysaccharide as immune enhancers, treating time and concentration of colchicines and so on. The result showed that 20% FBS with MEM and LPS (final concentration 0.3 mg/mL) was the best blood lymphocytic cell medium. Cultivating for 72h, treating the cells with colchicines (final concentration 0.08 μg/mL) for 3h before culture ending could get better chromosome samples. We also presented a comparison of several methods to identify the sex of the tonguefish,including the preparation of chromosome from cultured peripheral blood lym-phocytic cell, female-specific DNA marker, Physiological anatomy, gonadal biopsy and gonad cell culture. The first two methods could identify genetic sex of tonguefish exactly, while the other three methods could identify physical sex of tonguefish. However, there were big limitations of anatomical observation, Gonadal biopsy and gonadal cell culture. Preparation of blood lymphocyte chromosome and female-specific marker methods could

  18. Circulating human B and plasma cells. Age-associated changes in counts and detailed characterization of circulating normal CD138- and CD138+ plasma cells. : Blood B-lymphocytes and plasma cells in adults

    OpenAIRE

    2010-01-01

    International audience; Generation of B and plasma cells involves several organs with a necessary cell trafficking between them. A detailed phenotypic characterization of four circulating B-cell subsets (immature-, naïve-, memory- B-lymphocytes and plasma cells) of 106 healthy adults was realized by multiparametric flow cytometry. We show that CD10, CD27 and CD38 is the minimal combination of subsetting markers allowing unequivocal identification of immature (CD10(+)CD27(-)CD38(+), 6+/-6 cell...

  19. Massive ascites as a presenting manifestation of chronic lymphocytic leukemia

    Institute of Scientific and Technical Information of China (English)

    Neelam Siddiqui; Saeed Al-Amoudi; Aamer Aleem; Maha Arafah; Layla Al-Gwaiz

    2008-01-01

    Ascites is not an uncommon manifestation of certain solid tumors like gastrointestinal malignancies, ovarian cancer and breast cancer. However, it is unusual to encounter ascites in patients with hematological malignancies especially chronic leukemia. The patient described here presented with massive ascites and blood lymphocytosis. Further studies confirmed the diagnosis of chronic lymphocytic leukemia with ascites. The ascitic fluid was exudative, consisting of mature-looking B-lymphocytes, which were morphologically and immunophenotypically similar to peripheral blood and bone marrow cells. The patient was treated with chemotherapy and achieved a good response and diminution of ascitic fluid accumulation.

  20. Requirement for sialic acid on the endothelial ligand of a lymphocyte homing receptor

    OpenAIRE

    1990-01-01

    The entry of blood-borne lymphocytes into most secondary lymphoid organs is initiated by a highly specific adhesive interaction with the specialized cuboidal endothelial cells of high endothelial venules (HEV). The adhesive receptors on lymphocytes that dictate interactions with HEV in different lymphoid organs are called homing receptors, signifying their critical role in controlling organ-selective lymphocyte migration. Considerable work has established that the mouse peripheral lymph node ...