Sample records for blood leukocyte subsets

  1. Peripheral blood and milk leukocytes subsets of lactating Sarda ewes

    Piero Bonelli


    Full Text Available Leukocytes subpopulations in blood and milk of lactating Sarda ewes were investigated. Animals characterized by a SSC level <500×103cells/mL and a negative bacteriological examination were sampled in early, mid and late lactation. Milk differential cell count evidenced that macrophage represented the main population (42.8%±3.5 followed by lymphocytes (40.2%±3.4 and neutrophils (8,6%±2.1. Flow cytometry analysis showed that lymphocytes subsets in milk were quite different from blood. High CD8+ and low CD4+ lymphocytes percentages determined a CD4/CD8 ratio inversion in milk compared to blood (0.3%±0.03 vs 1.8%±0.08. CD8+ decreased while, conversely, CD4+ increased in late lactation. γδ T cells were more represented in milk (12.6%±1.3 than in blood (6.8%±0.3 and their proportions appeared similar throughout lactation in both compartments. IL-2 receptor was mainly expressed in milk on T cytotoxic lymphocytes. Data obtained in uninfected mammary glands could allow an early discrimination between physiological and pathological changes occurring in ewe milk. Further phenotypical and functional studies on milk leukocytes subsets might help to understand defense mechanisms of the ovine mammary gland against IMI.

  2. Ontogeny and characterization of blood leukocyte subsets and serum proteins in piglets before and after weaning

    Juul-Madsen, H.R.; Jensen, K.H.; Nielsen, Jens;


    Existing knowledge about the development of the porcine immune system was extended by phenotypic characterization of leukocyte subsets and with assessment of Mannan-Binding Lectin (MBL) and immunoglobulin concentrations in peripheral blood of healthy piglets. Single-color and/or double-color flow...... immune system seem to be stimulated immediately after weaning. At the time considered to have the highest infection pressure T-cells and TLR4+ cells were markedly enhanced, whereas the expression of SLA I did not seem to be affected by weaning....

  3. Social rank influences the distribution of blood leukocyte subsets in female growing pigs

    Hjarvard, Bodil Margrethe; Larsen, Ole Næsbye; Juul-Madsen, Helle Risdahl;


    The effect of high (DOM) and low (SUB) social rank on blood immune variables was examined in female growing pigs. Pigs were mixed with unfamiliar pigs at 9 weeks of age and kept in stable groups of 4 pigs for 5 weeks. Social rank was determined using a feeding competition test. SUB pigs showed...... reduced growth as compared to DOM pigs confirming their lower social status. Blood was sampled for immunological assessments immediately before grouping the pigs and again after the 5 weeks of social housing. White Blood Cell (WBC) counts, percentage of CD4 positive cells (CD4+), percentage of CD8...... positive cells (CD8+), percentage of swine leukocyte antigen II (SLAII) carrying cells, LPS-stimulated Toll-like Receptor 4 (TLR4) expression, and LPS-stimulated tumor necrosis factor-alpha (TNF-α) responsiveness were determined at both times. IgG and IgM concentrations were measured following the 5 weeks...

  4. CCR2 inhibition sequesters multiple subsets of leukocytes in the bone marrow.

    Fujimura, Naoki; Xu, Baohui; Dalman, Jackson; Deng, Hongping; Aoyama, Kohji; Dalman, Ronald L


    Chemokine receptor CCR2 mediates monocyte mobilization from the bone marrow (BM) and subsequent migration into target tissues. The degree to which CCR2 is differentially expressed in leukocyte subsets, and the contribution of CCR2 to these leukocyte mobilization from the BM are poorly understood. Using red fluorescence protein CCR2 reporter mice, we found heterogeneity in CCR2 expression among leukocyte subsets in varying tissues. CCR2 was highly expressed by inflammatory monocytes, dendritic cells, plasmacytoid dendritic cells and NK cells in all tissues. Unexpectedly, more than 60% of neutrophils expressed CCR2, albeit at low levels. CCR2 expression in T cells, B cells and NK T cells was greatest in the BM compared to other tissues. Genetic CCR2 deficiency markedly sequestered all leukocyte subsets in the BM, with reciprocal reduction noted in the peripheral blood and spleen. CCR2 inhibition via treatment with CCR2 signaling inhibitor propagermanium produced similar effects. Propagermanium also mitigated lipopolysaccharide-induced BM leukocyte egress. Consistent with its functional significance, CCR2 antibody staining revealed surface CCR2 expression within a subset of BM neutrophils. These results demonstrate the central role CCR2 plays in mediating leukocyte mobilization from the BM, and suggest a role for CCR2 inhibition in managing monocytes/macrophages-mediated chronic inflammatory conditions. PMID:26206182

  5. Crossing the Vascular Wall: Common and Unique Mechanisms Exploited by Different Leukocyte Subsets during Extravasation

    Michael Schnoor


    Full Text Available Leukocyte extravasation is one of the essential and first steps during the initiation of inflammation. Therefore, a better understanding of the key molecules that regulate this process may help to develop novel therapeutics for treatment of inflammation-based diseases such as atherosclerosis or rheumatoid arthritis. The endothelial adhesion molecules ICAM-1 and VCAM-1 are known as the central mediators of leukocyte adhesion to and transmigration across the endothelium. Engagement of these molecules by their leukocyte integrin receptors initiates the activation of several signaling pathways within both leukocytes and endothelium. Several of such events have been described to occur during transendothelial migration of all leukocyte subsets, whereas other mechanisms are known only for a single leukocyte subset. Here, we summarize current knowledge on regulatory mechanisms of leukocyte extravasation from a leukocyte and endothelial point of view, respectively. Specifically, we will focus on highlighting common and unique mechanisms that specific leukocyte subsets exploit to succeed in crossing endothelial monolayers.

  6. Margination of leukocytes in blood flow through small tubes.

    Goldsmith, H L; Spain, S


    Leukocyte margination in the vessels of the microcirculation has been attributed to a flow-dependent interaction with red cells. To determine the extent of this effect, experiments with human blood were done in 100- to 180-micron tubes to detect changes in cell distribution as a function of hematocrit and flow rate. Using a flow visualization technique, the leukocyte concentration distribution was determined in 45% ghost cell suspensions. Migration of cells toward the wall was observed at centerline velocities greater than 1 mm sec-1 and increased with increasing flow rate. The effect was probably due to a more rapid inward migration of ghosts than leukocytes because of fluid inertia and cell density differences. Experiments were therefore carried out in whole blood at hematocrits from 20 to 60%, measuring the number concentration of leukocytes and erythrocytes within the tube, nt, and comparing it to that in the infusing reservoir, no, (Fahraeus effect). At mean tube shear rates G less than 100 sec-1, nt/no less than 1 for both leukocytes and erythrocytes showing net migration of cells away from the wall, although at nearly all hematocrits there was an enrichment of leukocytes relative to erythrocytes in the tubes. At G less than 50 sec-1, nt/no remained less than 1 for erythrocytes but increased to greater than 1 for leukocytes showing migration toward the wall, the increase being greatest at 20% hematocrit in the 100-micron tubes. The nature of the effect was revealed by cine films which showed that, as the flow rate decreased, erythrocytes formed rouleaux which migrated inward creating a core and displacing leukocytes to the periphery. In control experiments using washed blood cells in phosphate buffer-albumin, nt/no less than 1 for both leukocytes and erythrocytes at all G and hematocrits, and leukocytes were now distributed. Cine films of washed blood confirmed that, in the absence of rouleaux, no significant inward migration of erythrocytes occurred. PMID

  7. Effect of transport stress on peripheral blood lymphocyte subsets and Th cytokines in pigs

    Wuren Ma


    Full Text Available In order to investigate transport stress on porcine peripheral blood lymphocyte subsets and Th cytokines, blood samples were collected from pigs before and after transport. Creatine kinase (CK, alkaline phosphatase (ALP, lactate dehydrogenase (LDH, glucose, and cortisol in the serum were measured. The number of leukocytes and lymphocytes, percentages of lymphocyte subsets, as well as Th cell cytokines level and their mRNA expression were detected, respectively. After transport, the level of CK, glucose and interleukin (IL-4 increased significantly (p<0.01, LDH, Th memory cells, natural killer and interferon (IFN-γ increased significantly (p<0.05, cortisol, number of leukocytes and lymphocytes decreased (p<0.01, percentages of γ δ T cells, naïve Th cells and cytotoxic T lymphocytes decreased significantly (p<0.05. The mRNA expressions IL-2 and IFN-γ were down regulated, p<0.01 and p<0.05, respectively. While IL-4, IL-6, and IL-10 were up regulated, but only IL-10 displayed a highly significant difference (p<0.01. These data suggested that transport could cause immune suppression in pigs, which influences cellular immunity more than humoral immunity, and humoral immunity may play an important role in transport stress. Proper measures should be taken to reduce susceptibility of infection after transport.

  8. Distinct single cell signal transduction signatures in leukocyte subsets stimulated with khat extract, amphetamine-like cathinone, cathine or norephedrine

    Bredholt, Therese; Ersvær, Elisabeth; Erikstein, Bjarte Skoe; Sulen, André; Reikvam, Håkon; Aarstad, Hans Jørgen; Johannessen, Anne Christine; Vintermyr, Olav Karsten; Bruserud, Øystein; Gjertsen, Bjørn Tore


    Background: Amphetamine and amphetamine derivatives are suggested to induce an immunosuppressive effect. However, knowledge of how amphetamines modulate intracellular signaling pathways in cells of the immune system is limited. We have studied phosphorylation of signal transduction proteins (Akt, CREB, ERK1/2, NF-κB, c-Cbl, STAT1/3/5/6) and stress sensors (p38 MAPK, p53) in human leukocyte subsets following in vitro treatment with the natural amphetamine cathinone, the cathinone d...

  9. DNA damage in peripheral blood leukocytes in tobacco users

    Venkateswara Rao Guttikonda; Rekha Patil; G S Kumar


    Aim : To Quantify the DNA single-stranded breaks in the peripheral blood leukocytes (PBLs) of tobacco-habituated individuals with clinically normal mucosa and patients with oral carcinoma. Objectives: To evaluate DNA damage levels in PBLs of tobacco-habituated individuals with clinically normal mucosa and patients with oral carcinoma and compare with a control group of healthy volunteers. To evaluate the extent of DNA damage in PBLs using Single Cell Gel Electrophoresis (SCGE) in the above gr...

  10. Leukocyte recovery from umbilical cord blood by poligeline.

    Perutelli, P; Catellani, S


    Umbilical cord blood (UCB) collected at delivery is a source of transplantable stem/progenitor cells; it represents an alternative to bone marrow to restore hematopoiesis in patients affected by malignant and non-malignant disease. Therefore, large-scale UCB banks would be a natural complement to bone marrow donor registries. Storage of unmanipulated whole UCB units requires a great number of liquid nitrogen containers. Separation of leukocytes allows UCB storage in smaller space, thus lowering banking costs; unfortunately, UCB processing may cause significant losses of stem cells. We report about the use of poligeline to remove erythrocytes from UCB units. After erythrocyte sedimentation at 1xg (30' or 40') or 50xg, leukocyte-rich supernatant was collected and centrifuged to recover the leukocyte pool in view of stem cell transplantation. Erythrocyte depletion was always satisfactory, ranging from 82.6% to 88.9%, but 1xg sedimentation for 40' enabled us to achieve the best CD34+ cell recovery (mean value 80.5%). The proposed UCB-processing method allowed us to lower the final sample volume down to 1/10 of the initial one, in this way making UCB banking feasible. Erythrocyte depletion took place directly in the collection bag, thus reducing microbial contamination risk. PMID:10193639

  11. Susceptibility and Response of Human Blood Monocyte Subsets to Primary Dengue Virus Infection

    Wong, Kok Loon; Chen, Weiqiang; Balakrishnan, Thavamalar; Toh, Ying Xiu; Fink, Katja; Wong, Siew-Cheng


    Human blood monocytes play a central role in dengue infections and form the majority of virus infected cells in the blood. Human blood monocytes are heterogeneous and divided into CD16− and CD16+ subsets. Monocyte subsets play distinct roles during disease, but it is not currently known if monocyte subsets differentially contribute to dengue protection and pathogenesis. Here, we compared the susceptibility and response of the human CD16− and CD16+ blood monocyte subsets to primary dengue viru...

  12. Multicolor flow cytometry analysis of blood cell subsets in patients given total body irradiation before bone marrow transplantation

    Bone marrow transplantation has often been closely linked with accidental or intentional therapeutical irradiation. In both situations, study of the radiosensitivity of human blood cell subsets is of interest. Using one-color flow cytometry analysis of B lymphocytes, T cell subsets, and natural killer cells, we previously reported that lymphocyte subsets exhibit equal radiosensitivity. Taking advantage of recent developments in the knowledge of leukocyte differentiation antigens and flow cytometry technology we undertook a study of blood cell subsets to search for rare populations exhibiting different radiosensitivity. Thirty patients, who were delivered a 12 Gy fractionated total body irradiation as part of their conditioning regimen before transplantation for malignant disorders, were studied using multicolor flow cytometry. T and B lymphocytes showed a sharp, radiation-induced decrease, with the B lymphocytes (cluster of differentiation (CD) 19+) being the most sensitive. When analyzed by multicolor flow cytometry all major lymphocyte subsets appeared equally sensitive to the in vivo irradiation. Therefore, all major lymphocyte subsets sharing the helper phenotype (naive or memory) and the cytotoxic phenotype appeared equally sensitive to in vivo whole body irradiation. In parallel, the CD34+ cell subset remained basically unchanged after whole body irradiation. Finally, the CD3-, 56+, 16+ natural killer cell subset was relatively radioresistant (91 and 74% of its initial value, after 2 and 4 Gy, respectively) as compared to other lymphocyte subsets. Our study provides evidence that T and B cell subsets seem to be highly radiosensitive in vivo. The CD34+ progenitor/stem cells and NK cells seem to be more radioresistant. This latter result might provide clues to the understanding of the pathophysiogeny of radiation-induced aplasia and of the engrafment/rejection process following bone marrow transplantation. 20 refs., 3 figs., 1 tab

  13. Multicolor flow cytometry analysis of blood cell subsets in patients given total body irradiation before bone marrow transplantation

    Purpose: Bone marrow transplantation has often been closely linked with accidental or intentional therapeutical irradiation. In both situations, study of the radiosensitivity of human blood cell subsets is of interest. Using one-color flow cytometry analysis of B lymphocytes, T cell subsets, and natural killer cells, we previously reported that lymphocyte subsets exhibit equal radiosensitivity. Taking advantage of recent developments in the knowledge of leukocyte differentiation antigens and flow cytometry technology we undertook a study of blood cell subsets to search for rare populations exhibiting different radiosensitivity. Methods and Materials: Thirty patients, who were delivered a 12 Gy fractionated total body irradiation as part of their conditioning regimen before transplantation for malignant disorders, were studied using multicolor flow cytometry. Results: T and B lymphocytes showed a sharp, radiation-induced decrease, with the B lymphocytes (cluster of differentiation (CD) 19+) being the most sensitive. When analyzed by multicolor flow cytometry, all major lymphocyte subsets appeared equally sensitive to the in vivo irradiation; that is, CD3+4+45RO+, CD3+4+45RA+, CD3+4+8-, CD3+4-8+. Therefore, all major lymphocyte subsets sharing the helper phenotype (naive or memory) and the cytotoxic phenotype appeared equally sensitive to in vivo whole body irradiation. In parallel, the CD34+ cell subset remained basically unchanged after whole body irradiation. Finally, the CD3-, 56+, 16+ natural killer cell subset was relatively radioresistant (91 and 74% of its initial value, after 2 and 4 Gy, respectively) as compared to other lymphocyte subsets. Conclusion: Our study provides evidence that T and B cell subsets seem to be highly radiosensitive in vivo. The CD34+ progenitor/stem cells and NK cells seem to be more radioresistant. This latter result might provide clues to the understanding of the pathophysiogeny of radiation-induced aplasia and of the engrafment

  14. Alteration of peripheral blood lymphocyte subsets in acute pancreatitis

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


    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.

  15. Effects of spray-dried porcine plasma and plant extracts on intestinal morphology and on leukocyte cell subsets of weaned pigs.

    Nofrarías, M; Manzanilla, E G; Pujols, J; Gibert, X; Majó, N; Segalés, J; Gasa, J


    We evaluated the effects of a 6% spray-dried porcine plasma (SDPP) and a plant extracts mixture (XT; 5% carvacrol, 3% cinnamaldehyde, and 2% capsicum oleoresin) on the productive performance, intestinal morphology, and leukocyte cell subsets of early-weaned pigs compared with a control group. Morphometry of the jejunum, ileum, and colon, and immune cell analysis of blood, ileocolic lymph node (LN), and ileal Peyer's patches were done in 24 weaned pigs (20 +/- 2 d) at 19 or 21 d postweaning. Although SDPP and XT treatments did not increase ADG or ADFI, SDPP improved the G:F ratio (P = 0.024) compared with the control group. Dietary SDPP reduced the percentages of blood monocytes (P = 0.006) and macrophages in ileal Peyer's patches and LN (P = 0.04), of B lymphocytes (P = 0.04) and gammadelta+ T cells in LN (P = 0.009), and of intraepithelial lymphocytes (P = 0.026) as well as the density of lamina propria cells in the colon (P < 0.01). Dietary XT reduced intraepithelial lymphocyte numbers in jejunum (P = 0.034) and the percentages of blood cytotoxic cells (P = 0.07) and B lymphocytes in LN (P = 0.03); however, XT increased blood monocytes (P = 0.038) and the density of lamina propria lymphocytes in the colon (P = 0.003). These results indicate that dietary SDPP and plant extracts can affect intestinal morphology and immune cell subsets of gut tissues and blood in weaned pigs. Furthermore, the effects of SDPP suggest lower activation of the immune system of the piglets. PMID:16971575

  16. Goat cathelicidin-2 is secreted by blood leukocytes regardless of lipopolysaccharide stimulation.

    Srisaikham, Supreena; Suksombat, Wisitiporn; Yoshimura, Yukinori; Isobe, Naoki


    It has been reported that goat cathelicidin-2, an antimicrobial peptide, localizes in leukocytes and is present in milk. Here, we examined whether cathelicidin-2 is secreted by leukocytes. Different concentrations (10(5) -10(8) cells/mL) of blood leukocytes were cultured for 0-48 h with or without lipopolysaccharide (LPS). After culture, the concentrations of cathelicidin-2 in the conditioned media were measured. Blood was collected from male goats 0-24 h after the intravenous injection of Escherichia coli O111:B4 LPS. The plasma cathelicidin-2 concentrations were determined and the blood leukocytes immunostained with anti-cathelicidin-2 antibody to calculate the proportion of cathelicidin-2-positive cells in the total leukocytes. When higher concentrations of leukocytes were cultured, the cathelicidin-2 concentrations in the media increased significantly, whereas the addition of LPS to the media caused no further increase. The plasma cathelicidin-2 concentrations did not increase with time after LPS infusion. The proportion of cathelicidin-2-positive cells in the total leukocytes was significantly reduced 1 h after LPS injection compared with that at 0 h, but increased again at 6 h and thereafter. These results suggest that cathlicidin-2 is secreted by leukocytes even without LPS stimulation, whereas LPS may be required for cathelicidin-2-containing leukocytes to be recruited from the blood to tissues showing inflammation. PMID:26212721

  17. The Effect of Hemiscorpius lepturus (Scorpionida: Hemiscorpiidae Venom on Leukocytes and the Leukocyte Subgroups in Peripheral Blood of Rat

    Mehri Ghafourian


    Full Text Available Background: The aim of this study was to investigate the effect of Hemiscorpius lepturus venom on leukocytes and the leukocyte subgroups in peripheral blood of rat.Methods: In this experimental study, sixty N-Mari rats were divided into three groups of 20 rats. Then the rats in each group were divided into four subgroups based on the blood sampling time that was 2, 6, 24 and 48 hours after the venom injection, respectively. The control group did not receive anything, however, the first and the second ex­perimental groups received 0.1 and 0.01mg/kg of venom, subcutaneously. In accordance with a designated four sam­pling times, the blood sampling was carried out in three groups. After RBC lysis, the leukocytes and leukocyte sub­populations were determined and counted using appropriate hematological standard methods.Results: The leukocyte and the neutrophil count at two (P<0.05, six (P<0.01 and 24 (P<0.05 hours after the venom injection showed a significant decline compared with the control group, this decrease was significant at the dose of 0.1 mg/kg until 48 hours after the venom injection (P<0.05. The lymphocyte count showed a significant decline throughout the all hours of the experiment, compared with the control group (P<0.05.Conclusion: Leukocytes are probably affected by the cytotoxicity effect of the H. lepturus venom in a dose-dependent manner. This could be a wakeup call for the medical staff to perform quick and accurate treatment in the least time possible.

  18. Production of fibrogenic cytokines by interleukin-2-treated peripheral blood leukocytes

    Kovacs, E J; Brock, B; Silber, I E;


    procollagen and fibronectin messenger RNAs was increased in human fibroblasts in response to leukocyte supernatants. Unstimulated leukocytes expressed minimal levels of transforming growth factor-beta or platelet-derived growth factor B chain messenger RNAs, but could be greatly enhanced by IL-2 treatment....... CONCLUSION: Mediators that induce connective tissue production are secreted by IL-2-treated peripheral blood leukocytes. These cytokines may be responsible, in part, for the stimulation of abdominal adhesions in patients receiving intraperitoneal immunotherapy....

  19. Shorter telomere length in peripheral blood leukocytes is associated with childhood autism

    Li, Zongchang; Tang, Jinsong; LI Hong; Chen, Shan; He, Ying; Liao, Yanhui; Wei, Zhen; Wan, Guobin; XIANG, XI; Xia, Kun; Chen, Xiaogang


    Telomeres are protective chromosomal structures that play a key role in preserving genomic stability. Epidemiologic studies have shown that the abnormal telomere length in leukocytes is associated with some mental disorders and age-related diseases. However, the association between leukocyte telomere length and autism has not been investigated. Here we investigated the possible association between relative telomere length (RTL) in peripheral blood leukocytes and childhood autism by using an e...

  20. Chronic inhalation of cigarette smoke reduces phagocytosis in peripheral blood leukocytes

    Tschernig, Thomas; Rabung, Andrea; Voss, Meike; Meier, Carola; Bals, Robert; Beisswenger, Christoph


    Background Phagocytosis activity of peripheral blood leukocytes in smokers or chronic obstructive pulmonary disease patients was found to be controversial and dependent on the phagocytic stimulus. Results We demonstrated that long-term exposure to cigarette smoke in mice clearly suppressed the phagocytosis of granulocytes and monocytes from peripheral blood. Conclusions Impaired phagocytosis activity of peripheral blood leukocytes may have a systemic effect and potentially contribute to smoki...

  1. Study of terahertz-radiation-induced DNA damage in human blood leukocytes

    Angeluts, A. A.; Gapeyev, A. B.; Esaulkov, M. N.; Kosareva, O. G.; Matyunin, S. N.; Nazarov, M. M.; Pashovkin, T. N.; Solyankin, P. M.; Cherkasova, O. P.; Shkurinov, A. P.


    We have carried out the studies aimed at assessing the effect of terahertz radiation on DNA molecules in human blood leukocytes. Genotoxic testing of terahertz radiation was performed in three different oscillation regimes, the blood leukocytes from healthy donors being irradiated for 20 minutes with the mean intensity of 8 - 200 μW cm-2 within the frequency range of 0.1 - 6.5 THz. Using the comet assay it is shown that in the selected regimes such radiation does not induce a direct DNA damage in viable human blood leukocytes.

  2. Study of terahertz-radiation-induced DNA damage in human blood leukocytes

    Angeluts, A A; Esaulkov, M N; Kosareva, O G; Solyankin, P M; Shkurinov, A P [International Laser Center, M. V. Lomonosov Moscow State University, Moscow (Russian Federation); Gapeyev, A B; Pashovkin, T N [Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Moscow Region (Russian Federation); Matyunin, S N [Section of Applied Problems at the Presidium of the Russian Academy of Sciences, Moscow (Russian Federation); Nazarov, M M [Institute on Laser and Information Technologies, Russian Academy of Sciences, Shatura, Moscow Region (Russian Federation); Cherkasova, O P [Institute of Laser Physics, Siberian Branch, Russian Academy of Sciences, Novosibirsk (Russian Federation)


    We have carried out the studies aimed at assessing the effect of terahertz radiation on DNA molecules in human blood leukocytes. Genotoxic testing of terahertz radiation was performed in three different oscillation regimes, the blood leukocytes from healthy donors being irradiated for 20 minutes with the mean intensity of 8 – 200 μW cm{sup -2} within the frequency range of 0.1 – 6.5 THz. Using the comet assay it is shown that in the selected regimes such radiation does not induce a direct DNA damage in viable human blood leukocytes. (biophotonics)

  3. A Leukocyte Immune-Type Receptor Subset Is a Marker of Antiviral Cytotoxic Cells in Channel Catfish, Ictalurus punctatus.

    Taylor, Erin B; Moulana, Mohadetheh; Stuge, Tor B; Quiniou, Sylvie M A; Bengten, Eva; Wilson, Melanie


    Channel catfish, Ictalurus punctatus, leukocyte immune type receptors (LITRs) represent a multigene family that encodes Ig superfamily proteins that mediate activating or inhibitory signaling. In this study, we demonstrate the use of mAb CC41 to monitor viral cytotoxic responses in catfish and determine that CC41 binds to a subset of LITRs on the surface of catfish clonal CTLs. Homozygous gynogenetic catfish were immunized with channel catfish virus (CCV)-infected MHC-matched clonal T cells (G14D-CCV), and PBL were collected at various times after immunization for flow cytometric analyses. The percentage of CC41(+) cells was significantly increased 5 d after primary immunization with G14D-CCV and at 3 d after a booster immunization as compared with control fish only injected with G14D. Moreover, CC41(+) cells magnetically isolated from the PBL specifically killed CCV-infected targets as measured by (51)Cr release assays and expressed messages for CD3γδ, perforin, and at least one of the CD4-like receptors as analyzed by RNA flow cytometry. When MLC effector cells derived from a G14D-CCV-immunized fish were preincubated with CC41 mAb, killing of G14D-CCV targets was reduced by ∼40%, suggesting that at least some LITRs have a role in target cell recognition and/or cytotoxicity. The availability of a LITR-specific mAb has allowed, to our knowledge for the first time, functional characterization of LITRs in an autologous system. In addition, the identification of an LITR subset as a cytotoxic cell marker will allow for more effective monitoring of catfish immune responses to pathogens. PMID:26856701

  4. Oligo-2',5'-adenylate synthetase activity in peripheral blood mononuclear leukocytes in various diseases.

    Fujii, N; Kotake, S.; Hirose, S; Ohno, S; Yasuda, I.; Sagawa, A; Ishikawa, K.; Minagawa, T


    Interferon induces oligo-2',5'-adenylate synthetase in cells. In various diseases, interferon was detectable in the circulation or was produced spontaneously from peripheral blood mononuclear leukocytes. The oligo-2',5'-adenylate synthetase activity in peripheral blood mononuclear leukocytes was examined in various diseases, including systemic lupus erythematosus, sarcoidosis, Vogt-Koyanagi-Harada disease, and Behcet's disease. The activity of this enzyme was significantly increased in system...

  5. Leukocytes Detection, Classification and Counting in Smears of Peripheral Blood

    Martínez-Castro, J.; Reyes-Cadena, S.; E. Felipe-Riverón


    Using the k -NN classifier in combination with the first Minkowski metric, in addition to techniques of digital image processing, we developed a computational system platform-independent, which is able to identify, to classify and to count five normal types of leukocytes: neutrophils, eosinophils, basophils, monocytes and lymphocytes. It is important to emphasize that this work does not attempt to differentiate between smears of leukocytes coming from healthy and sick people; this is because ...

  6. Report: Nuclei segmentation of leukocytes in blood smear digital images.

    Abbas, Naveed; Mohamad, Dzulkifli; Abdullah, Abdul Hanan; Saba, Tanzila; Al-Rodhaan, Mznah; Al-Dhelaan, Abdullah


    The Leukocytes are differentiated from each other on the basis of their nuclei, demanded in many Medical studies, especially in all types of Leukemia by the Hematologists to note the disorder caused by specific type of Leukocyte. Leukemia is a life threatening disease. The work for diagnosing is manually carried out by the Hematologists involving much labor, time and human errors. The problems mentioned are easily addressed through computer vision techniques, but still accuracy and efficiency are demanded in terms of the basic and challenging step segmentation of Leukocyte's nuclei. The underlying study proposed better method in terms of accuracy and efficiency by designing a dynamic convolution filter for boosting low intensity values in the separated green channel of an RGB image and suppressing the high values in the same channel. The high values in the green channel become 255 (background) while the nuclei always have low values in the green channel and thus clearly appear as foreground. The proposed technique is tested on 365 images achieving an overall accuracy of 95.89%, while improving the efficiency by 10%. The proposed technique achieved its targets in a realistic way by improving the accuracy as well as the efficiency and both are highly required in the area. PMID:26408877

  7. CD163 positive subsets of blood dendritic cells

    Maniecki, Maciej Bogdan; Møller, Holger Jon; Moestrup, Søren Kragh; Møller, Bjarne Kuno

    CD163 and CD91 are scavenging receptors with highly increased expression during the differentiation of monocytes into the anti-inflammatory macrophage phenotype. In addition, CD91 is expressed in monocyte-derived dendritic cells (MoDCs), where the receptor is suggested to be important for...... internalization of CD91-targeted antigens to be presented on the dendritic cell surface for T-cell stimulation. Despite their overlap in functionality, the expression of CD91 and CD163 has never been compared and the expression of CD163 in the monocyte-dendritic cell lineage is not yet characterized. CD163...... expression in dendritic cells (DCs) was investigated using multicolor flow cytometry in peripheral blood from 31 healthy donors and 15 HIV-1 patients in addition to umbilical cord blood from 5 newborn infants. Total RNA was isolated from MACS purified DCs and CD163 mRNA was determined with real-time reverse...

  8. Selection of the best features for leukocytes classification in blood smear microscopic images

    Sarrafzadeh, Omid; Rabbani, Hossein; Talebi, Ardeshir; Banaem, Hossein Usefi


    Automatic differential counting of leukocytes provides invaluable information to pathologist for diagnosis and treatment of many diseases. The main objective of this paper is to detect leukocytes from a blood smear microscopic image and classify them into their types: Neutrophil, Eosinophil, Basophil, Lymphocyte and Monocyte using features that pathologists consider to differentiate leukocytes. Features contain color, geometric and texture features. Colors of nucleus and cytoplasm vary among the leukocytes. Lymphocytes have single, large, round or oval and Monocytes have singular convoluted shape nucleus. Nucleus of Eosinophils is divided into 2 segments and nucleus of Neutrophils into 2 to 5 segments. Lymphocytes often have no granules, Monocytes have tiny granules, Neutrophils have fine granules and Eosinophils have large granules in cytoplasm. Six color features is extracted from both nucleus and cytoplasm, 6 geometric features only from nucleus and 6 statistical features and 7 moment invariants features only from cytoplasm of leukocytes. These features are fed to support vector machine (SVM) classifiers with one to one architecture. The results obtained by applying the proposed method on blood smear microscopic image of 10 patients including 149 white blood cells (WBCs) indicate that correct rate for all classifiers are above 93% which is in a higher level in comparison with previous literatures.

  9. Long telomeres in blood leukocytes are associated with a high risk of ascending aortic aneurysm.

    Tuija J Huusko

    Full Text Available Ascending aortic aneurysm is a connective tissue disorder. Even though multiple novel gene mutations have been identified, risk profiling and diagnosis before rupture still represent a challenge. There are studies demonstrating shorter telomere lengths in the blood leukocytes of abdominal aortic aneurysm patients. The aim of this study was to measure whether relative telomere lengths are changed in the blood leukocytes of ascending aortic aneurysm patients. We also studied the expression of telomerase in aortic tissue samples of ascending aortic aneurysms. Relative lengths of leukocyte telomeres were determined from blood samples of patients with ascending aortic aneurysms and compared with healthy controls. Telomerase expression, both at the level of mRNA and protein, was quantified from the aortic tissue samples. Mean relative telomere length was significantly longer in ascending aortic aneurysm blood samples compared with controls (T/S ratio 0.87 vs. 0.61, p<0.001. Expressions of telomerase mRNA and protein were elevated in the aortic aneurysm samples (p<0.05 and p<0.01. Our study reveals a significant difference in the mean length of blood leukocyte telomeres in ascending aortic aneurysm and controls. Furthermore, expression of telomerase, the main compensating factor for telomere loss, is elevated at both the mRNA and protein level in the samples of aneurysmal aorta. Further studies will be needed to confirm if this change in telomere length can serve as a tool for assessing the risk of ascending aortic aneurysm.

  10. Variation in sister chromatid exchange frequencies between human and pig whole blood, plasma leukocyte, and mononuclear leukocyte cultures

    Sister chromatid exchange (SCE) induction by ultraviolet (UV) light was studied in both human and pig whole blood cultures (WBC) and plasma leukocyte cultures (PLC). No variation in SCE frequency was observed between pig WBC and PLC in control as well as in treated cells. Conversely, SCE frequencies of human PLC were consistently higher than those of WBC in control and UV-exposed cells. Thus, red blood cells (RBCs) do not influence the sensitivity of lymphocytes to UV LIGHT exposure, and there must be some different culture condition(s) in the inducation of SCEs between human WBC and PLC but not in swine lymphocyte cultures. Since the BrdUrd/lymphocyte ratio of WBC was halved in PLC, the effect of BrdUrd concentration in inducing the SCE baseline frequency of PLC may be ruled out. Neither the cell separation technique nor polymorphonuclear leukocytes had a significant role in the elevated SCE frequency of human PLC or MLC. Experiments where human RBCs were titrated into human PLC showed that the induction of an elevated SCE frequency of PLC was suppressed in a dose-dependent manner by the presence of RBCs in the culture medium. Since the incorporation of pig or human RBCs into human PLC as well as into MLC reduced the SCE frequency to that of WBC, a common component and/or function existing in these cells is suggested. Analysis of different RBC components showed that RBCs, specifically RBC ghosts, release a diffusible but not dialyzable corrective factor into culture medium that is able to reduce the SCE frequencies of PLC

  11. Biomarkers measured in buccal and blood leukocyte DNA as proxies for colon tissue global methylation

    Ashbury, Janet E.; Taylor, Sherryl A; Tse, M Yat; Stephen C Pang; Louw, Jacob A; Vanner, Stephen J.; King, Will D


    There is increasing interest in clarifying the role of global DNA methylation levels in colorectal cancer (CRC) etiology. Most commonly, in epidemiologic studies, methylation is measured in DNA derived from blood leukocytes as a proxy measure of methylation changes in colon tissue. However, little is known about the correlations between global methylation levels in DNA derived from colon tissue and more accessible tissues such as blood or buccal cells. This cross-sectional study utilized DNA ...

  12. In vitro evaluation of canine leukocytes radiolabeled in whole blood with 99mTc stannous colloid

    Introduction: Technetium-99m stannous colloid (99mTcSnC)-labeled leukocytes are used to investigate a variety of inflammatory diseases in human medicine. The present study investigates the in vitro behavior of canine leukocytes labeled in whole blood with 99mTcSnC. Methods: Blood samples from 10 healthy dogs were labeled with 99mTcSnC using a standard procedure. The distribution of radioactivity among blood components (plasma, leukocyte layers and erythrocytes) was measured following separation of the radiolabeled samples across Histopaque density gradients. Phagocytic function of labeled and unlabeled leukocytes was estimated using zymosan particles. Labeling retention by leukocytes was determined at 1, 3, 4 and 7 h postlabeling. Results: The mean±standard error percentage of radioactivity associated with plasma, erythrocyte and leukocyte fractions was 2.0±0.21%, 55.5±0.60% and 42.5±0.54%, respectively (the last comprising 70.2±0.83% in polymorphonuclear leukocytes and 29.8±0.83% in mononuclear leukocytes). Labeled canine leukocytes had a phagocytic activity of 91.3±0.28% (control, 91.7±0.26%). The radiolabeled canine leukocytes retained 94.1±0.30% of radioactivity at 7 h postlabeling. Conclusions: Radiolabeling of canine leukocytes in whole blood with 99mTcSnC has minor adverse effect on their phagocytic function. The radiolabeled canine leukocytes retained a large percentage of radioactivity for at least 7 h postlabeling

  13. T-cell Subsets in Peripheral Blood and Tumors of Patients Treated With Oncolytic Adenoviruses

    Kristian, Taipale; Ilkka, Liikanen; Juuso, Juhila; Aila, Karioja-Kallio; Minna, Oksanen; Riku, Turkki; Nina, Linder; Johan, Lundin; Ari, Ristimäki; Anna, Kanerva; Anniina, Koski; Timo, Joensuu; Markus, Vähä-Koskela; Akseli, Hemminki


    The quality of the antitumor immune response is decisive when developing new immunotherapies for cancer. Oncolytic adenoviruses cause a potent immunogenic stimulus and arming them with costimulatory molecules reshapes the immune response further. We evaluated peripheral blood T-cell subsets of 50 patients with refractory solid tumors undergoing treatment with oncolytic adenovirus. These data were compared to changes in antiviral and antitumor T cells, treatment efficacy, overall survival, and T-cell subsets in pre- and post-treatment tumor biopsies. Treatment caused a significant (P < 0.0001) shift in T-cell subsets in blood, characterized by a proportional increase of CD8+ cells, and decrease of CD4+ cells. Concomitant treatment with cyclophosphamide and temozolomide resulted in less CD4+ decrease (P = 0.041) than cyclophosphamide only. Interestingly, we saw a correlation between T-cell changes in peripheral blood and the tumor site. This correlation was positive for CD8+ and inverse for CD4+ cells. These findings give insight to the interconnections between peripheral blood and tumor-infiltrating lymphocyte (TIL) populations regarding oncolytic virotherapy. In particular, our data suggest that induction of T-cell response is not sufficient for clinical response in the context of immunosuppressive tumors, and that peripheral blood T cells have a complicated and potentially misleading relationship with TILs. PMID:25655312

  14. Leukocyte subset analysis on blood from workers exposed to plutonium in 1944-45

    Monoclonal antibodies (MAbs) directed against cell surface markers were used to identify specific subpopulations of lymphocytes and monocytes obtained from 22 male subjects who had /sup 239/Pu exposures in 1944-45. Separated mononuclear cells were labelled simultaneously with fluoresceinated B1 (B cells), phycoerythrin Leu2a (T/sub S/ cells), LeμM3 with Texas-red goat-anti-mouse Ig and biotinylated Leμ3 with allophycocyanin avidin (T/sub H/ cells). In addition, the DNA-specific stain Hoechst 33342 was used to identify nucleated cells and to provide information on cell cycle status. Flow cytometric analysis using 3-color excitation (UV, 488 nm and 605 nm) was used, and 5 colors of emission were detected, corresponding to the four MAbs and Hoechst. The mononuclear cells were analyzed on the day of collection or after irradiation with 0, 50 or 200 rads followed by 6 days in culture with or without PHA stimulation. Marked differences in all measured parameters were found among the workers. The most striking response was a dose-related increase in the T/sub H//T/sub S/ ratio

  15. A rapid and simple method for the isolation of pure eosinophilic leukocytes from horse blood.

    Jörg, A; Portmann, P; Fellay, G; Dreyer, J L; Meyer, J


    An improved and short method is described for the isolation of intact eosinophilic leukocytes from horse blood with high yield (1--1.5 g/20 l). Viability and purity of the preparations were verified by light and electron microscopy and by the trypan blue exclusion test. Isolated eosinophils were 98--100% pure, intact and viable, and they could be shown to phagocytise immune-complexes. PMID:729750

  16. Effect of mixed γ-plus neutron-radiation on permeability to taurine of peripheral blood leukocyte membranes

    A study was made of permeability to taurine of cellular membranes of peripheral blood leukocytes in vitro under normal conditions and 24 k following mixed γ-plus neutron-irradiation in a dose of 3.5 Gy. It was established that radiation increases the taurine content of cells. The protein content of leukocytes also increases probably due to a better sorption of serum proteins of blood

  17. Leukocyte-subset counts in idiopathic parkinsonism provide clues to a pathogenic pathway involving small intestinal bacterial overgrowth. A surveillance study

    Dobbs R


    Full Text Available Abstract Background Following Helicobacter pylori eradication in idiopathic parkinsonism (IP, hypokinesia improved but flexor-rigidity increased. Small intestinal bacterial-overgrowth (SIBO is a candidate driver of the rigidity: hydrogen-breath-test-positivity is common in IP and case histories suggest that Helicobacter keeps SIBO at bay. Methods In a surveillance study, we explore relationships of IP-facets to peripheral immune/inflammatory-activation, in light of presence/absence of Helicobacter infection (urea-breath- and/or stool-antigen-test: positivity confirmed by gastric-biopsy and hydrogen-breath-test status for SIBO (positivity: >20 ppm increment, 2 consecutive 15-min readings, within 2h of 25G lactulose. We question whether any relationships found between facets and blood leukocyte subset counts stand in patients free from anti-parkinsonian drugs, and are robust enough to defy fluctuations in performance consequent on short t½ therapy. Results Of 51 IP-probands, 36 had current or past Helicobacter infection on entry, 25 having undergone successful eradication (median 3.4 years before. Thirty-four were hydrogen-breath-test-positive initially, 42 at sometime (343 tests during surveillance (2.8 years. Hydrogen-breath-test-positivity was associated inversely with Helicobacter-positivity (OR 0.20 (95% CI 0.04, 0.99, p In 38 patients (untreated (17 or on stable long-t½ IP-medication, the higher the natural-killer count, the shorter stride, slower gait and greater flexor-rigidity (by mean 49 (14, 85 mm, 54 (3, 104 mm.s-1, 89 (2, 177 Nm.10-3, per 100 cells.μl-1 increment, p=0.007, 0.04 & 0.04 respectively, adjusted for patient characteristics. T-helper count was inversely associated with flexor-rigidity before (p=0.01 and after adjustment for natural-killer count (-36(-63, -10 Nm.10-3 per 100 cells.μl-1, p=0.007. Neutrophil count was inversely associated with tremor (visual analogue scale, p=0.01. Effect-sizes were independent of IP

  18. HIV migration between blood plasma and cellular subsets before and after HIV therapy.

    Choi, Jun Yong; Chaillon, Antoine; Oh, Jin Ok; Ahn, Jin Young; Ann, Hae Won; Jung, In Young; Ahn, Mi-Young; Jeon, Yong Duk; Ku, Nam Su; Smith, Davey M; Kim, June Myung


    The cellular source of HIV RNA circulating in blood plasma remains unclear. Here, we investigated whether sequence analysis of HIV RNA populations circulating before combination antiretroviral therapy (cART) and HIV DNA populations in cellular subsets (CS) after cART could identify the cellular sources of circulating HIV RNA. Blood was collected from five subjects at cART initiation and again 6 months later. Naïve CD4+ T cells, resting central memory and effector memory CD4+ T cells, activated CD4+ T cells, monocytes, and natural killer cells were sorted using a fluorescence-activated cell sorter. HIV-1 env C2V3 sequences from HIV RNA in blood plasma and HIV DNA in CSs were generated using single genome sequencing. Sequences were evaluated for viral compartmentalization (Fst test) and migration events (MEs; Slatkin Maddison and cladistic measures) between blood plasma and each CS. Viral compartmentalization was observed in 88% of all cellular subset comparisons (range: 77-100% for each subject). Most observed MEs were directed from blood plasma to CSs (52 MEs, 85.2%). In particular, there was only viral movement from plasma to NK cells (15 MEs), monocytes (seven MEs), and naïve cells (five ME). We observed a total of nine MEs from activated CD4 cells (2/9 MEs), central memory T cells (3/9 MEs), and effector memory T cells (4/9 MEs) to blood plasma. Our results revealed that the HIV RNA population in blood plasma plays an important role in seeding various cellular reservoirs and that the cellular source of the HIV RNA population is activated central memory and effector memory T cells. J. Med. Virol. 88:606-613, 2016. © 2015 Wiley Periodicals, Inc. PMID:26348372

  19. Differences in the composition of the human antibody repertoire by B cell subsets in the blood

    Eva Szymanska eMroczek


    Full Text Available The vast initial diversity of the antibody repertoire is generated centrally by means of a complex series of V (D J gene rearrangement events, variation in the site of gene segment joining, and TdT catalyzed N- region addition. Although the diversity is great, close inspection has revealed distinct and unique characteristics in the antibody repertoires expressed by different B cell developmental subsets. In order to illustrate our approach to repertoire analysis, we present an in-depth comparison of V (D J gene usage, hydrophobicity, length, DH reading frame, and amino acid usage between heavy chain repertoires expressed by immature, transitional, mature, memory IgD+, memory IgD-, and plasmacytes isolated from the blood of a single individual. Our results support the view that in both human and mouse the H chain repertoires expressed by individual, developmental B cell subsets appear to differ in sequence content. Sequencing of unsorted B cells from the blood is thus likely to yield an incomplete or compressed view of what is actually happening in the immune response of the individual. Our findings support the view that studies designed to correlate repertoire expression with diseases of immune function will likely require deep sequencing of B cells sorted by subset.

  20. Focal MMP-2 and MMP-9 Activity at the Blood-Brain Barrier Promotes Chemokine-Induced Leukocyte Migration

    Jian Song


    Full Text Available Although chemokines are sufficient for chemotaxis of various cells, increasing evidence exists for their fine-tuning by selective proteolytic processing. Using a model of immune cell chemotaxis into the CNS (experimental autoimmune encephalomyelitis [EAE] that permits precise localization of immigrating leukocytes at the blood-brain barrier, we show that, whereas chemokines are required for leukocyte migration into the CNS, additional MMP-2/9 activities specifically at the border of the CNS parenchyma strongly enhance this transmigration process. Cytokines derived from infiltrating leukocytes regulate MMP-2/9 activity at the parenchymal border, which in turn promotes astrocyte secretion of chemokines and differentially modulates the activity of different chemokines at the CNS border, thereby promoting leukocyte migration out of the cuff. Hence, cytokines, chemokines, and cytokine-induced MMP-2/9 activity specifically at the inflammatory border collectively act to accelerate leukocyte chemotaxis across the parenchymal border.

  1. rhG-CSF in healthy donors: mobilization of peripheral hemopoietic progenitors and effect on peripheral blood leukocytes.

    Sica, S; Rutella, S; Di Mario, A; Salutari, P; Rumi, C; Ortu la Barbera, E; Etuk, B; Menichella, G; D'Onofrio, G; Leone, G


    Recombinant human granulocyte colony-stimulating factor (rhG-CSF) 16 micrograms/kg/day was given to 9 healthy donors to recruit hemopoietic progenitors (HP) for allogeneic transplantation or donor leukocyte infusion. rhG-CSF was administered s.c. for 5 days. No side effects were encountered except for moderate bone pain and lumbago. Mobilization was effective, reaching a peak median value of 187 x 10(3) CD34+ cells/ml (range 51.2-1127) and 2170 x 10(3) colony-forming units-granulocyte macrophage (CFU-GM)/ml (range 1138-4190). Peak values were obtained at a median of 4 days of rhG-CSF and represented, respectively, a 13-fold and a 37-fold increase from baseline values (p = 0.0007 and p = 0.006). White blood cell (WBC) counts increased 6-fold from baseline values (p < 0.0007) and reached a median peak of 34 x 10(6)/ml (23.5-59). Polymorphonuclear (PMN), and mononuclear (MNC) cells increased 10-fold and 2-fold, respectively (p = 0.0039 and p = 0.0026) and reached a median peak of 32.1 x 10(6)/ml (18.2-52) and 4.42 x 10(6)/ml (3.14-12.42). Absolute lymphocyte and monocyte counts increased at peak day in all donors 1.5-fold and 5.7-fold from baseline values (p = 0.0017 and p = 0.0018). In 7 of 9 donors, lymphocyte subsets were analyzed in detail. CD3+ and CD19+ lymphocytes increased 1.5-fold and 3-fold, respectively (p = 0.032 for both). NK and activated T lymphocytes doubled at a median of 4 days of rhG-CSF (p = 0.032 and p = NS, respectively). Similar changes were observed in lymphocytes collected in leukapheresis product. T helper and T suppressor subsets displayed a similar increase. Thus, besides the anticipated priming effect on HP and PMN, rhG-CSF in healthy donors produced an unexpected and still unexplained modification of lymphocyte subsets in peripheral blood. PMID:8877714

  2. Transfusion of leukocyte-depleted red blood cells is not a risk factor for nosocomial infections in critically ill children

    van der Wal, Judith; van Heerde, Marc; Markhorst, Dick G.; Kneyber, Martin C. J.


    Objectives: Transfusion of red blood cells is increasingly linked with adverse outcomes in critically ill children. We tested the hypothesis that leukocyte-depleted red blood cell transfusions were independently associated with increased development of bloodstream infections, ventilator-associated p

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

    顾军; 范清源; 高春芳; 代夫; 郑茂荣


    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+.

  4. Multicentre evaluation of stable reference whole blood for enumeration of lymphocyte subsets by flow cytometry.

    Edwards, Cherry; Belgrave, Danielle; Janossy, George; Bradley, Nicholas J; Stebbings, Richard; Gaines-Das, Rose; Thorpe, Robin; Sawle, Alex; Arroz, Maria Jorge; Brando, Bruno; Gratama, Jan Willem; Orfao de Matos, Alberto; Papa, Stephano; Papamichail, Michael; Lenkei, Rodica; Rothe, Gregor; Barnett, David


    BACKGROUND: Clinical indications for lymphocyte subset enumeration by flow cytometry include monitoring of disease progression and timing of therapeutic intervention in infection with human immunodeficiency virus. Until recently international standardisation has not been possible due to a lack of suitable stable reference material. METHODS: This study consisted of two trials of a stabilised whole blood preparation. Eleven participants were sent two standard protocols for staining plus gating strategy and asked to report absolute counts for lymphocyte subsets. RESULTS: No significant difference was detected between the two methods when results from the two assays and all partners were pooled. Significant differences in results from the different partners were observed. However, representative mean counts were obtained for geometric means, geometric coefficient of variation, and 95% confidence interval for CD3 910 cells/mul, 9%, and 888 to 933, respectively), CD4 (495 cells/mul, 12%, and 483 to 507), and CD8 (408 cells/mul, 13%, and 393 to 422). CONCLUSION: We have introduced a stabilised blood preparation and a well-characterized biological standard. The availability of this reference material greatly simplifies the validation of new techniques for CD4(+) T-cell enumeration and the expansion of external quality assurance programmes for clinical laboratories, including those that operate in resource-restricted environments. (c) 2005 Wiley-Liss, Inc. PMID:15973699

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

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


    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.

  6. Immunophenotype characteristics of peripheral blood mononuclear leukocytes of chronic idiopathic urticaria patients.

    Garmendia, Jenny V; Zabaleta, Mercedes; Aldrey, Oscar; Rivera, Henry; De Sanctis, Juan B; Bianco, Nicolás E; Blanca, Isaac


    The pathogenesis of chronic idiopathic urticaria (CIU) is not completely understood although autoimmunity has been proposed. The aim of the study was to assess the expression of different leukocyte antigens, by flow cytometry, assaying total blood of 29 patients with CIU and of 20 sex and age matched controls. Moreover, we assessed soluble CD154 a marker of immune cell activation, predominantly memory T cells. When patients were divided depending an their response to the autologous serum skin test (ASST), three different groups were encountered: group 1 (n=11): with negative ASST-, group 2 (n=11): positive ASST (ASST+) with normal lymphocyte counts and group 3 (n=7): ASST+ with low lymphocyte counts (CIU pathogenesis. PMID:17176904

  7. [Comparative transcriptome analysis of human aorta atherosclerotic lesions and peripheral blood leukocytes from essential hypertension patients].

    Timofeeva, A V; Goriunova, L E; Khaspekov, G L; Il'inskaia, O P; Sirotkin, V N; Andreeva, E R; Tararak, E M; Bulkina, O S; Buza, V V; Britareva, V V; Karpov, Iu A; Bibilashvili, R Sh


    One of the major cardiovascular risk factor which predisposes to and accelerates atherosclerosis is arterial hypertension (AH). To determine the molecular basis of the crosslink between AH and atherosclerosis for the development of new treatment strategies large-scale transcriptome analysis of the cells implicated in atherogenesis is needed. We used cDNA microarray technique for simultaneous analysis of gene expression in human abdominal aorta normal sites and atherosclerotic lesions of different histological types, as well as in peripheral blood leukocytes from patients with essential hypertension (EH) and donors. The microarray data were verified by quantitative RT-PCR (reverse transcription coupled with polymerase chain reaction) and immunohistochemical analysis. Differential expression of 40 genes has been found, among which twenty two genes demonstrated up-regulation and 18 genes demonstrated down-regulation in atherosclerotic aorta compared with normal vessel. New gene-candidates, implicated in atherogenesis, have been identified - FPRL2, CD37, CD53, RGS1, LCP1, SPI1, CTSA, EPAS1, FHL1, GEM, RHOB, SPARCL1, ITGA8, PLN, and COL14A1. These genes participate in cell migration and adhesion, phenotypic changes of smooth muscle cells, immune and inflammatory reactions, oxidative processes and extracellular matrix remodeling. We have found increased expression levels of CD53, SPI1, FPRL2, SPP1, CTSD, ACP5, LCP1, CTSA and LIPA genes in peripheral blood leukocytes from EH patients and in atherosclerotic lesions of human aorta. The majority of these genes significantly (p0.5) correlated with AH stage as well as with histological grading of atherosclerotic lesions. PMID:19772500

  8. Age gene expression and coexpression progressive signatures in peripheral blood leukocytes.

    Irizar, Haritz; Goñi, Joaquín; Alzualde, Ainhoa; Castillo-Triviño, Tamara; Olascoaga, Javier; Lopez de Munain, Adolfo; Otaegui, David


    Both cellular senescence and organismic aging are known to be dynamic processes that start early in life and progress constantly during the whole life of the individual. In this work, with the objective of identifying signatures of age-related progressive change at the transcriptomic level, we have performed a whole-genome gene expression analysis of peripheral blood leukocytes in a group of healthy individuals with ages ranging from 14 to 93 years. A set of genes with progressively changing gene expression (either increase or decrease with age) has been identified and contextualized in a coexpression network. A modularity analysis has been performed on this network and biological-term and pathway enrichment analyses have been used for biological interpretation of each module. In summary, the results of the present work reveal the existence of a transcriptomic component that shows progressive expression changes associated to age in peripheral blood leukocytes, highlighting both the dynamic nature of the process and the need to complement young vs. elder studies with longitudinal studies that include middle aged individuals. From the transcriptional point of view, immunosenescence seems to be occurring from a relatively early age, at least from the late 20s/early 30s, and the 49-56 year old age-range appears to be critical. In general, the genes that, according to our results, show progressive expression changes with aging are involved in pathogenic/cellular processes that have classically been linked to aging in humans: cancer, immune processes and cellular growth vs. maintenance. PMID:26362218

  9. Effects of spaceflight on rat peripheral blood leukocytes and bone marrow progenitor cells

    Ichiki, A. T.; Gibson, L. A.; Jago, T. L.; Strickland, K. M.; Johnson, D. L.; Lange, R. D.; Allebban, Z.


    The white blood cell (WBC) elements and the bone marrow myeloid progenitor cell populations were analyzed to ascertain adaptation to micro-gravity and subsequent readaptation to 1 G in rats flown on the 14-day Spacelab Life Sciences-2 (SLS-2) mission. Bone marrow cells were harvested from one group of rats killed inflight (FD13) and blood was drawn from three other groups at various times. The WBC level was normal on FD14 with the exception of neutrophilia. On FD13, numbers of colony-forming units-granulocyte (CFU-G), CFU-GM, and CFU-M from flight animals were decreased compared with ground controls when incubated with recombinant rat interleukin-3 (rrIL-3) alone or in combination with recombinant human erythropoietin (rhEpo). On recovery (R + 0), flight rats had decreased numbers of total leukocytes and absolute numbers of lymphocytes and monocytes with elevated neutrophils compared with control rats. They had lower numbers of CD4, CD8, CD2, CD3, and B cells in the peripheral blood but no differences in spleen lymphocytes.

  10. Methylation of a panel of genes in peripheral blood leukocytes is associated with colorectal cancer.

    Luo, Xiang; Huang, Rong; Sun, Hongru; Liu, Yupeng; Bi, Haoran; Li, Jing; Yu, Hongyuan; Sun, Jiamei; Lin, Shangqun; Cui, Binbin; Zhao, Yashuang


    The relationship between the DNA methylation status of the CpG islands of multiple genes in blood leukocytes in CRC susceptibility and prognosis, as well as possible interactions with dietary factors on CRC risk are unclear. We carried out a case-control study including 421 CRC patients and 506 controls to examine the associations between six genes (AOX-1, RARB2, RERG, ADAMTS9, IRF4, and FOXE-1), multiple CpG site methylation (MCSM) and susceptibility to CRC. High-level MCSM (MCSM-H) was defined as methylation of greater than or equal to 2 of 5 candidate genes (except for RARB2); low-level MCSM (MCSM-L) was when 1 candidate gene was methylated; non-MCSM was when none of the candidate genes were methylated. Blood cell-derived DNA methylation status was detected using methylation-sensitive high-resolution melting analysis. The hypermethylation status of each individual gene was statistically significantly associated with CRC. MCSM status was also associated with CRC (OR = 1.54, 95% CI: 1.15-2.05, P = 0.004). We observed interactions between a high level of dietary intake of cereals, pungent food, and stewed fish with brown sauce, age (older than 60 yrs), smoking and hypermethylation on risk of CRC. MCSM in peripheral blood DNA may be an important biomarker for susceptibility to CRC. PMID:27453436

  11. Dysregulation of CXCR3 expression on peripheral blood leukocytes in patients with neovascular age-related macular degeneration

    Falk, Mads Krüger; Singh, Amardeep; Faber, Carsten; Nissen, Mogens H; Hviid, Thomas; Sørensen, Torben Lykke


    Purpose: The chemokine receptor CXCR3 has been strongly related to inhibition of angiogenesis. The purpose of this study was to investigate the association between expression of CXCR3 on peripheral blood leukocytes and Age-related Wet Macular Degeneration (AMD). Furthermore, we measured the plasma...

  12. Similar blood-borne DNA methylation alterations in cancer and inflammatory diseases determined by subpopulation shifts in peripheral leukocytes

    Li, H; Zheng, T.; Chen, B; Hong, G; Zhang, W.; Shi, T; Li, S.; Ao, L; Wang, C.; Guo, Z.


    Background: Although many DNA methylation (DNAm) alterations observed in peripheral whole blood/leukocytes and serum have been considered as potential diagnostic markers for cancer, their origin and their specificity for cancer (e.g., vs inflammatory diseases) remain unclear. Methods: From publicly available datasets, we identified changes in the methylation of blood-borne DNA for multiple cancers and inflammatory diseases. We compared the identified changes with DNAm difference between myelo...

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


    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.

  14. Comparison of quantitative HCMV DNAemia in whole blood and leukocytes, and Real-Time PCR in a BMT patient

    Elio De Nisco


    Full Text Available Background. HCMV is the most opportunistic viral agent in the transplantation of bone marrow and solid organ. Early therapy based on the detection of HCMV pp65 antigen in peripheral blood leukocytes, has led to a significant reduction in the incidence of related HCMV diseases.The pp65 antigenemia is difficult to standardize, while the quantification of HCMV DNA by Real-Time PCR is an alternative diagnostic approach with greater sensitivity and reproducibility, providing important information in the management of infected patients. Objectives. The aim of this study was the comparative analysis of quantitative HCMV DNAemia in leukocytes and in whole blood, with Real-time PCR, in a BMT patient with HCMV post-transplant reactivation, in order to analyze the relationship between levels of viral DNA at different time-points obtained in the two biological matrices. Study Design. The presence of HCMV DNA was detected in whole blood and peripheral blood leukocytes in a patient who underwent allogeneic marrow transplant for Ph1 + acute lymphoblastic leukemia in molecular remission. After 5-6 months, the patient has increased molecular Bcr-Abl (Philadelphia chromosome. It was activated immune reaction by means of tapering (lowering the dose of cyclosporine that uses the GvL effect to turn negative the Philadelphia chromosome positivity (Bcr/Abl negativity. Later, the patient develops GvHD and cortisone is administered. The persistence of grafts treated with immunosuppressants periodically reactivates HCMV infection. The DNA extraction from whole-blood was performed by automatic extractor QIACUBE (Qiagen, while extraction from leucocytes was performed on a standard number of leukocytes (EXTRAcell- Nanogen.The extracted DNA was amplified by Real-Time Alert Q-PCR (Nanogen.The samples were analyzed weekly for about 5 months from 1 year after transplantation. Results. The patient at 1 year after transplantation, has three HCMV reactivation at 56th, 62th and 67

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

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


    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...

  16. Establishment of a microplate assay for flow cytometric assessment and it is use for the evaluation of age-related phenotypic changes in canine whole blood leukocytes.

    Reis, Alexandre B; Carneiro, Cláudia M; Carvalho, Maria das Graças; Teixeira-Carvalho, Andréa; Giunchetti, Rodolfo C; Mayrink, Wilson; Genaro, Odair; Corrêa-Oliveira, Rodrigo; Martins-Filho, Olindo A


    The effectiveness of flow cytometric assays for canine use is still requiring standardization. Despite several studies using purified mononuclear cells, no methodology or reference ranges are available for immunophenotyping of whole blood leukocytes (WBL). Fresh and pre-fixed WBL were used to identify cell-subsets, (Thy-1(+)/CD5(+)/CD4(+)/CD8(+)/CD21(+) and CD14(+)) and measure MHC-II, CD45RA/CD45RB expression. We described here an efficient method for fast quantification of canine-WBL, using pre-fix in a microplate assay, which allows long-term sample storage prior to phenotyping. Decreased percentage of CD5(+)-T-cells within the lymphocyte-gate and increased percentage of CD21(+)-B-cells were observed in young animals, which led to higher T/B cell ratios in middle-aged dogs. Lower numerical counts of Thy-1(+), CD4(+), CD8(+) and CD21(+) lymphocyte were observed when compared to young animals. In addition, we identified an age-related decline of MHC-II/CD45RA expression by lymphocytes. We proposed an improved method for phenotyping of canine peripheral blood mononuclear cells (PBMC) that has significant use for researchers and veterinary clinicians. The hematological changes of senescence previously identified on PBMC could be adequately reproduced on features identified by whole blood. Furthermore, this study supplies normal range references as baseline standards for clinical purposes, besides specific immunological parameters to monitor canine aging process. PMID:15621304

  17. Reactions of peripheral blood mononuclear cells (PBMC) of camels with monoclonal antibodies against ruminant leukocytes.

    Ungar-Waron, H; Yagil, R; Brenner, J; Paz, R; Partosh, N; Van Creveld, C; Lubashevsky, E; Trainin, Z


    The particular immune system of the camel has been but little investigated. In this work circulating camel peripheral blood mononuclear cells (PBMC) were studied by flow cytometry. Monoclonal antibodies (mAbs) raised against ruminant leukocytes were used for the detection of cell surface antigens. Monoclonals to T-cell markers, CD4 (CACT138A) and CD8 (CACT80C), exhibited no reactivity towards camel PBMC in contrast to their reactivity to PBMC of other ruminant species and those of cattle in particular. A relatively high percentage (29.1+/-8.9%) of camel PBMC reacted with a non-immunoglobulin cell surface marker, B-B2, comparable to the reactivity of bovine PBMC. The B-B7 cell marker revealed 22.4+/-10.0% of reactive camel PBMC while the CD45 leukocyte common antigen was identified only on 19.4+/-3.1% of camel PBMC as compared to 74.7+/-4.9% for bovine PBMC. IgM (PIg45A) was detected on 9.1+/-1.4% of camel PBMC and on 46.6+/-19.5% of the bovine PBMC. Double fluorescent labeling with two B-cell markers and an anti-ruminant lambda light-chain mAb revealed 7-9% of cells bearing both B and lambda L-chain markers. Light chain reactivity was also assessed using an anti-goat F(ab')(2) antiserum. The values obtained, 14.3+/-5.8% for the camel and 47.8+/-2.7% for the cattle, are close to the values observed for surface IgM. These data suggest that camels, like other ruminants, possess L-chain bearing cells of the B-cell lineage. However, in the camel, Igs are different in that in addition to regular four chain Igs, about 65% of them possess two heavy chain Igs devoid of light chains. Because different sets of V(H) gene segments are used by four and two chain Igs, it is possible that there might be two lineages of B-cells each secreting a different form of antibodies. PMID:12493494

  18. A novel thyroid stimulating hormone β-subunit isoform in human pituitary, peripheral blood leukocytes, and thyroid

    Schaefer, Jeremy S.; Klein, John R.


    Thyroid stimulating hormone (TSH) is produced by the anterior pituitary and is used to regulate thyroid hormone output, which in turn controls metabolic activity. Currently, the pituitary is believed to be the only source of TSH used by the thyroid. Recent studies in mice from our laboratory have identified a TSH isoform that is expressed in the pituitary, in peripheral blood leukocytes (PBL), and in the thyroid. To determine whether a human TSH splice variant exists that is analogous to the ...

  19. Biotin Deficiency Reduces Expression of SLC19A3, a Potential Biotin Transporter, in Leukocytes from Human Blood12

    Vlasova, Tatyana I; Stratton, Shawna L.; Wells, Amanda M.; Mock, Nell I.; Mock, Donald M.


    In evaluating potential indicators of biotin status, we quantitated the expression of biotin-related genes in leukocytes from human blood of normal subjects before and after inducing marginal biotin deficiency. Biotin deficiency was induced experimentally by feeding an egg-white diet for 28 d. Gene expression was quantitated for the following biotin-related proteins: methylcrotonyl-CoA carboxylase chains A (MCCA) and B (MCCB); propionyl-CoA carboxylase chains A (PCCA) and B (PCCB); pyruvate c...

  20. Cryopreservation of blood mononuclear leukocytes and stem cells suspended in a large fluid volume. A preclinical model for a blood stem cell bank.

    Fliedner, T M; Körbling, M; Calvo, W; Bruch, C; Herbst, E


    It was the purpose of this study to establish and evaluate a freezing-and-thawing method for preservation of hemopoietic stem cells from the peripheral blood. Blood leukocytes collected by means of an IBM Blood-Cell-Separator were frozen in plastic bags using 10% DMSO and controlled cooling rates. Thawing was performed rapidly, and DMSO was diluted and removed prior to the in-vitro and in-vivo assays. The mean recovery of mononuclear cells collected from 82 leukaphereses was 86%. To assess the recovery of cryopreserved hemopoietic stem cells, the soft agar culture method adapted for the dog was used. There was no significant difference in the CFUc recovery per 1 X 10(6) mononuclear cells or in per leukapheresis after different cryopreservation times (1--6 and 7--27 months). To evaluate the hemopoietic repopulation capability of cryopreserved blood stem cells, leukapheresis-derived leukocytes were transfused into 1200 R whole body x-irradiated dogs. The hemopoietic repopulation pattern at day 10 after transfusion of comparable numbers of fresh or frozen leukocytes was not significantly different, as measured in bone marrow smears and sections and by granulocyte concentration in the peripheral blood. PMID:912104

  1. Amifostine (WR2721) Confers DNA Protection to In Vivo Cisplatin-Treated Murine Peripheral Blood Leukocytes

    Prieto González, E. A.; Fuchs, A. G.; Sánchez, González S.


    Amifostine [S-2-3-aminopropil amino ethyl phosphorotioic acid], a modulator agent for antineoplastic drugs involved in free radicals generation has given controversial results in cisplatin treated leukocytes in vitro. We have evaluated the amifostine protection over leukocytes in vivo, using comet assay. Groups of five OF1 male mice were given one of three doses of amifostine (56, 105 and 200 mg/Kg) after a cisplatin single injection (10 mg/Kg). Serum malonyldialdehide levels, catalase and su...

  2. Effect of low dose irradiation on subsets of T-lymphocyte of peripheral blood, spleen and tumor tissue

    Purpose: In order to understand the mechanism of the stimulation effects of low dose radiation (LDR), the author observed the immune changes of T-lymphocyte subsets. Meteria and methods: Whole body of BALB/C bring-tumor mice were exposed to the doses of 5, 10, 20 and 50 cGy γ-rays. The changes of T-lymphocyte subsets in peripheral blood, spleen and tumor-infiltrating lymphocyte (TIL) were studied with flow cytometry (FCM). Results: the ratio of L3T4+/Lyt2+ remarkable increased in the peripheral blood and spleen (p3T4+/Lyt2+ further decreased in the TIL group of mice exposed 10 cGy (p2+ molecules, were concentrated in the tumor tissues and they carried out the killing function to the tumor cells

  3. Genotoxic differences by sex in nasal epithelium and blood leukocytes in subjects residing in a highly polluted area

    We describe differences by sex in genotoxic damage found in a population of medical students exposed to a highly oxidative atmosphere, compared with a control group, measured by the single-cell gel electrophoresis assay and histological changes in nasal epithelium smears. Cells were obtained from the nasal epithelium and blood leukocytes. Higher DNA damage in nasal cells and leukocytes was found in males compared to females and control subjects. The percentage of squamous metaplastic changes in the nasal epithelium was also higher in males compared with females and controls. The commutation of normal nasal epithelium by squamous cells might modify its protective function in the nose, increasing the risk of damage to the lower respiratory tract. Although, as medical students, males and females were exposed to the same environment and activity patterns, male genotoxicity damage was higher in control and exposed subjects. More research should be done in order to identify direct or indirect sexual hormone intervention

  4. The modulating effect of royal jelly consumption against radiation-induced apoptosis in human peripheral blood leukocytes

    Navid Rafat


    Full Text Available The present work was designed to assess the radioprotective effect of royal jelly (RJ against radiation-induced apoptosis in human peripheral blood leukocytes. In this study, peripheral blood samples were obtained on days 0, 4, 7, and 14 of the study from six healthy male volunteers taking a 1000 mg RJ capsule orally per day for 14 consecutive days. On each sampling day, all collected whole blood samples were divided into control and irradiated groups which were then exposed to the selected dose of 4 Gy X-ray. Percentage of apoptotic cells (Ap % was evaluated for all samples immediately after irradiation (Ap0 and also after a 24 h postirradiation incubation at 37°C in 5% CO2 (Ap24 by the use of neutral comet assay. Concerning Ap0, collected data demonstrated that the percentage of apoptotic cells in both control and irradiated groups did not significantly change during the study period. However, with respect to Ap24, the percentage of apoptotic cells in irradiated groups gradually reduced during the experiment, according to which a significant decrease was found after 14 days RJ consumption (P = 0.002. In conclusion, the present study revealed the protective role of 14 days RJ consumption against radiation-induced apoptosis in human peripheral blood leukocytes.

  5. Correlation of MLH1 and MGMT methylation levels between peripheral blood leukocytes and colorectal tissue DNA samples in colorectal cancer patients



    CpG island methylation in the promoter regions of the DNA mismatch repair gene mutator L homologue 1 (MLH1) and DNA repair gene O6-methylguanine-DNA methyltransferase (MGMT) genes has been shown to occur in the leukocytes of peripheral blood and colorectal tissue. However, it is unclear whether the methylation levels in the blood leukocytes and colorectal tissue are correlated. The present study analyzed and compared the levels of MGMT and MLH1 gene methylation in the leukocytes of peripheral blood and colorectal tissues obtained from patients with colorectal cancer (CRC). The methylation levels of MGMT and MLH1 were examined using methylation-sensitive high-resolution melting (MS-HRM) analysis. A total of 44 patients with CRC were selected based on the MLH1 and MGMT gene methylation levels in the leukocytes of the peripheral blood. Corresponding colorectal tumor and normal tissues were obtained from each patient and the DNA methylation levels were determined. The correlation coefficients were evaluated using Spearman’s rank test. Agreement was determined by generalized κ-statistics. Spearman’s rank correlation coefficients (r) for the methylation levels of the MGMT and MLH1 genes in the leukocytes of the peripheral blood and normal colorectal tissue were 0.475 and 0.362, respectively (P=0.001 and 0.016, respectively). The agreement of the MGMT and MLH1 gene methylation levels in the leukocytes of the peripheral blood and normal colorectal tissue were graded as fair and poor (κ=0.299 and 0.126, respectively). The methylation levels of MGMT and MLH1 were moderately and weakly correlated between the patient-matched leukocytes and the normal colorectal tissue, respectively. Blood-derived DNA methylation measurements may not always represent the levels of normal colorectal tissue methylation. PMID:24179526

  6. Differences in the composition of the human antibody repertoire by B cell subsets in the blood

    Eva Szymanska eMroczek; Ippolito, Gregory C.; Tobias eRogosch; Kam Hon eHoi; Tracy A Hwangpo; Marsha G Brand; Yingxin eZhuang; Cun Ren eLiu; Schneider, David A; Michael eZemlin; Brown, Elizabeth E.; George eGeorgiou; Schroeder, Harry W.


    The vast initial diversity of the antibody repertoire is generated centrally by means of a complex series of V (D) J gene rearrangement events, variation in the site of gene segment joining, and TdT catalyzed N- region addition. Although the diversity is great, close inspection has revealed distinct and unique characteristics in the antibody repertoires expressed by different B cell developmental subsets. In order to illustrate our approach to repertoire analysis, we present an in-depth com...

  7. Differences in the Composition of the Human Antibody Repertoire by B Cell Subsets in the Blood

    Mroczek, Eva Szymanska; Ippolito, Gregory C.; Rogosch, Tobias; Hoi, Kam Hon; Tracy A Hwangpo; Marsha G Brand; Zhuang, Yingxin; Liu, Cun Ren; Schneider, David A; Zemlin, Michael; Brown, Elizabeth E.; Georgiou, George; Schroeder, Harry W.


    The vast initial diversity of the antibody repertoire is generated centrally by means of a complex series of V(D)J gene rearrangement events, variation in the site of gene segment joining, and TdT catalyzed N-region addition. Although the diversity is great, close inspection has revealed distinct and unique characteristics in the antibody repertoires expressed by different B cell developmental subsets. In order to illustrate our approach to repertoire analysis, we present an in-depth comparis...

  8. Peripheral Blood Lymphocyte Subsets in Adolescents: a Longitudinal Analysis from the REACH Project

    Bret J. Rudy; Wilson, Craig M.; Durako, Stephen; Moscicki, Anna-Barbara; Muenz, Larry; Douglas, Steven D.


    Flow cytometry analysis of lymphocyte subset markers was performed for a group of sexually active, human immunodeficiency virus (HIV)-negative adolescents over a 2-year period to establish normative data. Data were collected in the REACH Project (Reaching for Excellence in Adolescent Care and Health), a multicenter, longitudinal study of HIV-positive and high-risk HIV-negative adolescents. Two- and three-color flow cytometry data were collected every 6 months for these subjects. We determined...

  9. Defining mononuclear phagocyte subset homology across several distant warm-blooded vertebrates through comparative transcriptomics

    Thien eVu Manh


    Full Text Available Mononuclear phagocytes are organized in a complex system of ontogenically and functionally-distinct subsets, that has been best described in mouse and to some extent in human. Identification of homologous mononuclear phagocyte subsets in other vertebrate species of biomedical, economic and environmental interest is needed to improve our knowledge in physiologic and physio-pathologic processes, and to design intervention strategies against a variety of diseases, including zoonotic infections.We developed a streamlined approach combining refined cell sorting and integrated comparative transcriptomics analyses which revealed conservation of the mononuclear phagocyte organization across human, mouse, sheep, pigs and, in some respect, chicken. This strategy should help democratizing the use of omics analyses for the identification and study of cell types across tissues and species. Moreover we identified conserved gene signatures that enable robust identification and universal definition of these cell types. We identified new evolutionarily conserved gene candidates and gene interaction networks for the molecular regulation of the development or functions of these cell types, as well as conserved surface candidates for refined subset phenotyping throughout species. A phylogenetic analysis revealed that orthologous genes of the conserved signatures exist in teleost fishes and apparently not in Lamprey, indicating conservation of the genetic support for mononuclear phagocyte organization throughout jawed vertebrates but likely not in agnathans. Altogether this work provides molecular clues to the definition and functions of mononuclear phagocyte subsets across vertebrates which shall be useful to rigorously identify these cells and to design universal strategies to manipulate them in many target species towards the goal to reach and maintain global health.

  10. Influence of surgical treatment and radiotherapy of the advanced intraoral cancers on complete blood count, body mass index, liver enzymes and leukocyte CD64 expression:

    Didanovič, Vojislav; Dovšak, Tadej; Ihan Hren, Nataša; Ihan, Alojz; Kansky, Andrej


    Background. The aim of our study was to evaluate the influence of the surgery and radiotherapy of the advanced oral squamous cell carcinoma on the complete blood count, body mass index (BMI), acute inflam-matory response, liver enzymes and expression of the CD64 index on leukocytes in the peripheral blood. Patients and method. Venous blood was obtained from 16 patients with advanced oral squamous cell carcinomas treated with radical surgery and external beam radiotherapy. Blood samples were c...

  11. Global DNA hypomethylation in peripheral blood leukocytes as a biomarker for cancer risk: a meta-analysis.

    Hae Dong Woo

    Full Text Available BACKGROUND: Good biomarkers for early detection of cancer lead to better prognosis. However, harvesting tumor tissue is invasive and cannot be routinely performed. Global DNA methylation of peripheral blood leukocyte DNA was evaluated as a biomarker for cancer risk. METHODS: We performed a meta-analysis to estimate overall cancer risk according to global DNA hypomethylation levels among studies with various cancer types and analytical methods used to measure DNA methylation. Studies were systemically searched via PubMed with no language limitation up to July 2011. Summary estimates were calculated using a fixed effects model. RESULTS: The subgroup analyses by experimental methods to determine DNA methylation level were performed due to heterogeneity within the selected studies (p<0.001, I(2: 80%. Heterogeneity was not found in the subgroup of %5-mC (p = 0.393, I(2: 0% and LINE-1 used same target sequence (p = 0.097, I(2: 49%, whereas considerable variance remained in LINE-1 (p<0.001, I(2: 80% and bladder cancer studies (p = 0.016, I(2: 76%. These results suggest that experimental methods used to quantify global DNA methylation levels are important factors in the association study between hypomethylation levels and cancer risk. Overall, cancer risks of the group with the lowest DNA methylation levels were significantly higher compared to the group with the highest methylation levels [OR (95% CI: 1.48 (1.28-1.70]. CONCLUSIONS: Global DNA hypomethylation in peripheral blood leukocytes may be a suitable biomarker for cancer risk. However, the association between global DNA methylation and cancer risk may be different based on experimental methods, and region of DNA targeted for measuring global hypomethylation levels as well as the cancer type. Therefore, it is important to select a precise and accurate surrogate marker for global DNA methylation levels in the association studies between global DNA methylation levels in peripheral

  12. Isolation of Human Skin Dendritic Cell Subsets.

    Gunawan, Merry; Jardine, Laura; Haniffa, Muzlifah


    Dendritic cells (DCs) are specialized leukocytes with antigen-processing and antigen-presenting functions. DCs can be divided into distinct subsets by anatomical location, phenotype and function. In human, the two most accessible tissues to study leukocytes are peripheral blood and skin. DCs are rare in human peripheral blood (Nestle et al., J Immunol 151:6535-6545, 1993). These factors led to the extensive use of skin DCs as the "prototype" migratory DCs in human studies. In this chapter, we detail the protocols to isolate DCs and resident macrophages from human skin. We also provide a multiparameter flow cytometry gating strategy to identify human skin DCs and to distinguish them from macrophages. PMID:27142012

  13. The impact of HIV infection on blood leukocyte responsiveness to bacterial stimulation in asymptomatic patients and patients with bloodstream infection

    Huson, Michaëla A M; Hoogendijk, Arie J; de Vos, Alex F; Grobusch, Martin P; van der Poll, Tom


    Introduction HIV-induced changes in cytokine responses to bacteria may influence susceptibility to bacterial infections and the consequent inflammatory response. Methods We examined the impact of HIV on whole blood responsiveness to bacterial stimulation in asymptomatic subjects and patients with bacterial bloodstream infection (BSI). Whole blood was stimulated ex vivo with two bacterial Toll-like receptor agonists (lipopolysaccharide and lipoteichoic acid) and two pathogens (Streptococcus pneumoniae and non-typhoidal Salmonella), which are relevant in HIV-positive patients. Production of interferon-γ, tumour necrosis factor-α, interleukin-1β and interleukin-6 was used as a read-out. Results In asymptomatic subjects, HIV infection was associated with reduced interferon-γ, release after stimulation and priming of the pro-inflammatory cytokine response to non-typhoidal Salmonella. In patients with BSI, we found no such priming effect, nor was there evidence for more profound sepsis-induced immunosuppression in BSI patients with HIV co-infection. Conclusions These results suggest a complex effect of HIV on leukocyte responses to bacteria. However, in patients with sepsis, leukocyte responses were equally blunted in patients with and without HIV infection. PMID:27189532


    I. D. Bespalovа


    Full Text Available The aim of this study was to assess effects of the eight-week course of atorvastatin therapy upon the levels of spontaneous cytokine production by mononuclear blood leukocytes (MNBC in metabolic syndrome. An open-label prospective study included 36 patients with stage II hypertension (blood pressure < 180/110 mm Hg. accomplished by metabolic syndrome. Along with clinical surveys performed at a specialized cardiological clinics, we assessed spontaneous cytokine production by MNBC during treatment with atorvastatin. It was shown that the 8-week treatment of these patients with atorvaststin, at individually matched daily doses (20to 40 mg was associated with reduced serum concentration of acute phase proteins (C-reactive protein and neopterin, as well as decreased spontaneous production of proinflammatory cytokines (IL-1β, IL-6 and TNFα by MNBCs. The latter finding is of great importance for pathogenesis of metabolic syndrome.

  15. Development of methods to examine the effects of atmospheric particulate matter (PM) on human peripheral blood leukocytes

    Zussman, Lisa Ann

    In vitro methods to study the effect of atmospheric particulate matter (PM) on leukocyte function using human peripheral blood were developed. These methods were demonstrated using the blood of 1-5 individuals and National Institute of Standards and Technology (NIST) urban PM #1648, diesel PM #1650, silica PM, and a locally collected PM sample (New Jersey PM10). For the blood samples analyzed in this study NIST urban PM and New Jersey PM10 treatment mediated the release of granule contents from peripheral blood leukocytes and induced structural changes associated with degranulation. Flow cytometry revealed PM-induced changes in phagocytosis and cell structure associated with degranulation. Transmission electron microscopy confirmed NIST urban PM-induced cell structure changes were associated with PM internalization. Colorametric and electrophoretic methods showed no PM-induced release of primary granules and a slight PM-induced release of secondary granules associated with only NIST urban PM. Enzyme Immunosorbent Assays detected increased histamine release from basophils treated with NIST urban PM, a locally collected PM, and the soluble and insoluble components of these particles. NIST urban PM was found to be a potent inducer of histamine release in 4 out of 6 individuals tested. Fractionation studies revealed that soluble (aqueous) and insoluble fractions of NIST urban PM contain histamine-releasing activity. This was also demonstrated for the New Jersey PM10 sample for which the soluble fraction exhibited the most activity. Complementary studies with inhibitors of IgE-mediated histamine release conducted on one test subject suggest that PM-induced histamine release was partially mediated by IgE. A new hypothesis has been formed, suggesting that particle toxicity is related to PM-induced histamine release. Due to the bioactive nature of histamine and its association with many cardiopulmonary responses, the PM- mediated release of histamine should be investigated

  16. Effects of low-dose radiation on T lymphocyte subsets in peripheral blood of different positions of male radiation doctor

    Objective: To investigate effects of the ionizing radiation on T lymphocyte subsets in peripheral blood of different positions of male radiation personnel. Methods: The CD3+T, CD3+/CD4+T, CD3+/CD8+T, CD4+/CD45RA+T, CD4+/CD45RO+T, CD8+/CD45RA+T, CD8+/CD45RO+T, CD8+/CD28+T, CD8+/CD95+T, CD8+/HLA-DR+T percentage in the peripheral blood lymphocyte of 168 male radiation personnel were tested and compared by different jobs and working ages, with 39 healthy male controls. Results: In CT group, CD3+/CD8+T between ∼20 and 21∼ working ages groups was statistically significant difference (t=-2.07, P+/CD8+T and CD8+/CD45RO+T between CT group and control group were both significant different (t=-2.04, P+/CD8+T lymphocyte subset was mainly caused by low-dose ionizing radiation in male radiation doctor who worked in CT position longer than 20 years. We should pay attention to that. (authors)

  17. The impact of maternal HIV infection on cord blood lymphocyte subsets and cytokine profile in exposed non-infected newborns

    Reis-Alves Suiellen C


    Full Text Available Abstract Background Children born to HIV+ mothers are exposed intra-utero to several drugs and cytokines that can modify the developing immune system, and influence the newborn's immune response to infections and vaccines. We analyzed the relation between the distribution of cord blood lymphocyte subsets and cytokine profile in term newborns of HIV+ mothers using HAART during pregnancy and compared them to normal newborns. Methods In a prospective, controlled study, 36 mother-child pairs from HIV+ mothers and 15 HIV-uninfected mothers were studied. Hematological features and cytokine profiles of mothers at 35 weeks of pregnancy were examined. Maternal and cord lymphocyte subsets as well as B-cell maturation in cord blood were analyzed by flow cytometry. The non-stimulated, as well as BCG- and PHA-stimulated production of IL2, IL4, IL7, IL10, IL12, IFN-γ and TNF-alpha in mononuclear cell cultures from mothers and infants were quantified using ELISA. Results After one year follow-up none of the exposed infants became seropositive for HIV. An increase in B lymphocytes, especially the CD19/CD5+ ones, was observed in cord blood of HIV-exposed newborns. Children of HIV+ hard drug using mothers had also an increase of immature B-cells. Cord blood mononuclear cells of HIV-exposed newborns produced less IL-4 and IL-7 and more IL-10 and IFN-γ in culture than those of uninfected mothers. Cytokine values in supernatants were similar in infants and their mothers except for IFN-γ and TNF-alpha that were higher in HIV+ mothers, especially in drug abusing ones. Cord blood CD19/CD5+ lymphocytes showed a positive correlation with cord IL-7 and IL-10. A higher maternal age and smoking was associated with a decrease of cord blood CD4+ cells. Conclusions in uninfected infants born to HIV+ women, several immunological abnormalities were found, related to the residual maternal immune changes induced by the HIV infection and those associated with antiretroviral

  18. Measurement of lymphocyte subsets in peripheral blood of groups of workers in Kozloduy NPP

    The assessment of health risk is one of the key problems in the radiobiology nowadays. The damage of the immune system, following radiation exposure, can result in unwilling health consequences. The aim of the present study was to elucidate the effect of low dose of ionizing radiation on the cellular immunity of occupational exposed persons. 186 individuals from Kozloduy NPP, age 29-62 years were involved in the study. The workers were divided in the following groups: 1) Electricity production enterprise (1 and 2) - 48 persons with cumulated doses above 200 mSv and 40 persons with cumulated doses below 200 mSv; 2) Operating staff - 38 persons with cumulated doses about 11 mSv; 3) Managing staff - 30 persons with cumulated doses about 41 mSv, 4) Plant control group - 30 persons. The lymphocytes subsets were measured by flow cytometry, using a FacScan analyzer and immunofluorescent staining mouse monoclonal antibodies Becton Dickinson, USA, production. In present study the immunological parameters of Kozloduy NPP workers generally stayed within normal limits. Correlation coefficients to cumulated dose, age and working experience were low. However for some parameters, apparently within the assumed normal range, the differences in the results between the group with cumulated doses below 200 mSv or that of operating staff compared to the plant control group were statistically significant. It seemed reasonable to conclude that low doses radiation did not influence the cellular immunity of NPP workers seriously, although we could not exclude that more obvious effects might evolve later. All this gave us motivation to extend and continue studies of occupational exposed people

  19. Long-term impact of puerperal metritis on the profiles of peripheral blood leukocytes in peripartum dairy cows.

    Magata, Fumie; Kitaoka, Ryuji; Morino, Ikumi; Teramura, Makoto; Kawashima, Chiho; Haneda, Shingo; Shimizu, Takashi


    To determine the effects of puerperal metritis on the immune response, changes in the differential peripheral blood leukocyte counts were analyzed during the peripartum period in cows with or without metritis. Multiparous Holstein cows were examined for uterine health disorders and classified into two groups: healthy (n = 11) or metritis (n = 5) cows. The lymphocyte and monocyte counts and the proportion of CD8(+) lymphocytes were higher in cows with metritis compared to healthy cows. Moreover, the effects of puerperal metritis on the lymphocyte counts and CD4(+) /CD8(+) ratio persisted weeks after the uterine inflammation had self-resolved. Taken together, the findings of the present study indicate the possible long-term alterations of systemic immune responses in cows with puerperal uterine inflammation. © 2015 Japanese Society of Animal Science. PMID:26387573

  20. Genome-wide peripheral blood leukocyte DNA methylation microarrays identified a single association with inflammatory bowel diseases

    Harris, R Alan; Nagy-Szakal, Dorottya; Pedersen, Natalia; Opekun, Antone; Bronsky, Jiri; Munkholm, Pia; Jespersgaard, Cathrine; Andersen, PaalSkytt; Melegh, Bela; Ferry, George; Jess, Tine; Kellermayer, Richard


    Crohn's disease (CD) and ulcerative colitis (UC) are common forms of inflammatory bowel disease (IBD). Monozygotic (MZ) twin discordance rates and epidemiologic data implicate that environmental changes and epigenetic factors may play a pathogenic role in IBD. DNA methylation (the methylation of ...... cytosines within CpG dinucleotides) is an epigenetic modification, which can respond to environmental influences. We investigated whether DNA methylation might be connected with IBD in peripheral blood leukocyte (PBL) DNA by utilizing genome-wide microarrays.......Crohn's disease (CD) and ulcerative colitis (UC) are common forms of inflammatory bowel disease (IBD). Monozygotic (MZ) twin discordance rates and epidemiologic data implicate that environmental changes and epigenetic factors may play a pathogenic role in IBD. DNA methylation (the methylation of...

  1. Molecular signatures induced by interleukin-2 on peripheral blood mononuclear cells and T cell subsets

    Stroncek David


    Full Text Available Experimentally, interleukin-2 (IL-2 exerts complex immunological functions promoting the proliferation, survival and activation of T cells on one hand and inducing immune regulatory mechanisms on the other. This complexity results from a cross talk among immune cells which sways the effects of IL-2 according to the experimental or clinical condition tested. Recombinant IL-2 (rIL-2 stimulation of peripheral blood mononuclear cells (PBMC from 47 donors of different genetic background induced generalized T cell activation and anti-apoptotic effects. Most effects were dependent upon interactions among immune cells. Specialized functions of CD4 and CD8 T cells were less dependent upon and often dampened by the presence of other PBMC populations. In particular, cytotoxic T cell effector function was variably affected with a component strictly dependent upon the direct stimulation of CD8 T cells in the absence of other PBMC. This observation may provide a roadmap for the interpretation of the discrepant biological activities of rIL-2 observed in distinct pathological conditions or treatment modalities.

  2. Leukocyte count affects expression of reference genes in canine whole blood samples

    Piek, C.J.; Brinkhof, B.; Rothuizen, J.; Dekker, A.; Penning, L.C.


    Background The dog is frequently used as a model for hematologic human diseases. In this study the suitability of nine potential reference genes for quantitative RT-PCR studies in canine whole blood was investigated. Findings The expression of these genes was measured in whole blood samples of 263 i

  3. Degeneration and atrophy of the thymus of lethally irradiated dogs, rescued by transfusion of cryopreserved autologous blood leukocytes

    Dogs exposed to a fatal radiation dose of 12 Gy were rescued by transfusion of autologous blood leukocytes. A severe acute and long-lasting damage to the thymus was observed. The acute damage, as observed on the tenth day, consisted of a marked reduction in the number of lymphocytes, degeneration of Hassall's bodies, and hemorrhage. Long-term effects, observed several months after irradiation, were partial to total atrophy of the thymus. Regeneration, when it occurred, was limited to a few small isolated areas in which lymphopoiesis was supported by epithelial reticular cells. In contrast, the lymph nodes of all dogs had abundant cortical lymphopoiesis. The abundant hemopoiesis present in the marrow from the tenth day after irradiation until the end of the observation period should have provided sufficient circulating precursor cells to seed the thymus and regenerate the organ to the same extent as that observed in the other blood-forming organs. The impairment of lymphopoietic regeneration in the thymus seems to be due, therefore, to damage caused by irradiation on the specific stroma of the organ, which is not able to support such activity

  4. [Use of leukocyte-filtered, cytomegalovirus-antibody negative and irradiated cellular blood products].

    Solheim, B G; Albrechtsen, D H; Evensen, S A; Leivestad, T


    This paper presents both quality requirements and indications for use of leucocyte-filtered, cytomegalovirus antibody negative and irradiated cellular blood products at Rikshospitalet. Emphasis is placed on the use of standardized buffycoat depleted red cells or platelet concentrates for filtration, and the selection of leucocyte filters with high capacity and ease of bedside application. Leucocyte counts as low as 1-2 10(5) per unit are recommended after filtration in order to avoid HLA-antibody production. For bedside filtration, our choice was RC100 and PL100 (Pall) for red cells and platelets respectively. For laboratory use we prefer, for economic reasons, to use Sepacell R500 (Asahi) and Imugard IG500 (Terumo) for red cells and platelets respectively. Leucocyte-filtered blood products are considered indicated in all pre-transplant transfusions, in post-transplant HLA-sensitized patients, in other patients with febrile transfusion reactions, and in patients with an expected protracted platelet requirement. CMV antibody negative products are recommended for all immuno-deficient patients and pregnant women negative for CMV antibody. Irradiated blood products are used in the treatment of immuno-deficient patients receiving large amounts of blood, and in all severely immuno-compromised patients. In emergency situations where CMV antibody negative and/or irradiated blood products cannot be supplied, leucocyte filtration is suggested. PMID:2154061

  5. Changes of human leukocyte phagocytic activity after ultraviolet irradiation of blood

    Phagocytic activity of human mono- and granulocytes increased markedly after UV blood irradiation in the apparatus ''Izolda'' used in hospitals of the USSR for medical treatment. With the rise of irradiation dose the ratio of cells ingesting latex particles increased, although the average number of particles ingested per cell decreased. The integrative phagocytic index poorly depended on the irradiation dose. In patients with a low initial level of phagocytic index, after UV blood irradiation it became more pronounced than in those with the initial elevated level. The enhancement of phagocytic activity is the result of a direct UV-stimulation of cells. This stimulation not mediated by irradiated blood plasma is known to inhibit the phagocytic activity of leucocytes. A possible mechanism of phagocytic activity stimulation is discussed discussed

  6. Leukocyte count affects expression of reference genes in canine whole blood samples

    Dekker Aldo


    Full Text Available Abstract Background The dog is frequently used as a model for hematologic human diseases. In this study the suitability of nine potential reference genes for quantitative RT-PCR studies in canine whole blood was investigated. Findings The expression of these genes was measured in whole blood samples of 263 individual dogs, representing 73 different breeds and a group of 40 mixed breed dogs, categorized into healthy dogs and dogs with internal and hematological diseases, and dogs that underwent a surgical procedure. GeNorm analysis revealed that a combination of 5 to 6 of the most stably expressed genes constituted a stable normalizing factor. Evaluation of the expression revealed different ranking of reference genes in Normfinder and GeNorm. The disease category and the white blood cell count significantly affected reference gene expression. Conclusions The discrepancy between the ranking of reference genes in this study by Normfinder and Genorm can be explained by differences between the experimental groups such as "disease category" and "WBC count". This stresses the importance of assessing the expression stability of potential reference genes for gene experiments in canine whole blood anew for each specific experimental condition.

  7. Changes in some Blood Micronutrients, Leukocytes and Neutrophil Expression of Adhesion Molecules in Periparturient Dairy Cows

    Petersson L


    Full Text Available Dairy cows are highly susceptible to infectious diseases, like mastitis, during the period around calving. Although factors contributing to increased susceptibility to infection have not been fully elucidated, impaired neutrophil recruitment to the site of infection and changes in the concentrations of some micronutrients related with the function of the immune defence has been implicated. Most of the current information is based on studies outside the Nordic countries where the conditions for dairy cows are different. Therefore, the aim of the study was to evaluate changes in blood concentrations of the vitamins A and E, the minerals calcium (Ca, phosphorous (P, and magnesium (Mg, the electrolytes potassium (K and sodium (Na and the trace elements selenium (Se, copper (Cu and zinc (Zn, as well as changes in total and differential white blood cell counts (WBC and expression of the adhesion molecules CD62L and CD18 on blood neutrophils in Swedish dairy cows during the period around calving. Blood samples were taken from 10 cows one month before expected calving, at calving and one month after calving. The results were mainly in line with reports from other countries. The concentrations of vitamins A and E, and of Zn, Ca and P decreased significantly at calving, while Se, Cu, and Na increased. Leukocytosis was detected at calving, mainly explained by neutrophilia, but also by monocytosis. The numbers of lymphocytes tended to decrease at the same time. The mean fluorescent intensity (MFI of CD62L and CD18 molecules on blood neutrophils remained constant over time. The proportion of CD62L+ neutrophils decreased significantly at calving. The animals were fed according to, or above, their requirements. Therefore, changes in blood levels of vitamins, minerals and trace elements were mainly in response to colostrum formation, changes in dry matter intake, and ruminal metabolism around calving. Decreased levels of vitamins A and E, and of Zn at calving

  8. Human Blood CXCR5+CD4+ T Cells Are Counterparts of T Follicular Cells and Contain Specific Subsets that Differentially Support Antibody Secretion

    Morita, Rimpei; Schmitt, Nathalie; Bentebibel, Salah-Eddine; Ranganathan, Rajaram; Bourdery, Laure; Zurawski, Gerard; Foucat, Emile; Dullaers, Melissa; Oh, SangKon; Sabzghabaei, Natalie; Lavecchio, Elizabeth M; Punaro, Marilynn; Pascual, Virginia; Banchereau, Jacques; Ueno, Hideki


    Although a fraction of human blood memory CD4+ T cells expresses chemokine (C-X-C motif) receptor 5 (CXCR5), their relationship to T follicular helper (Tfh) cells is not well-established. Here we show that human blood CXCR5+ CD4+ T cells share functional properties with Tfh cells, and appear to represent their circulating memory compartment. Blood CXCR5+ CD4+ T cells comprised three subsets; T helper 1 (Th1), Th2 and Th17 cells. Th2 and Th17 cells within CXCR5+, but not within CXCR5−, compartment efficiently induced naïve B cells to produce immunoglobulins via interleukin-21 (IL-21). In contrast, Th1 cells from both CXCR5+ and CXCR5− compartments lacked the capacity to help B cells. Patients with juvenile dermatomyositis, a systemic autoimmune disease, displayed a profound skewing of blood CXCR5+ Th subsets towards Th2 and Th17 cells. Importantly, the skewing of subsets correlated with disease activity and frequency of blood plasmablasts. Collectively, our study suggests that an altered balance of Tfh subsets contributes to human autoimmunity. PMID:21215658

  9. Labelling of T cell subsets under field conditions in tropical countries. Adaptation of the immuno-alkaline phosphatase staining method for blood smears

    Lisse, I M; Whittle, H; Aaby, P;


    Immuno-alkaline phosphatase (AP) staining for T cell subsets (CD4 and CD8) of smears from fingerprick blood functioned well under tropical climatic conditions when smears were stored frozen with silica gel before being labelled. Unlabelled smears were stored for up to 12 months and could be...

  10. Postnatal development of leukocyte subsets composition

    Toman, M.; Faldyna, M.; Knotigová, P.; Pokorová, D.; Šinkora, Jiří

    Uppsala: International Union of Immunological Societies, 2001. s. 43. [International Veterinary Immunology Symposium /6./. 15.07.2001-20.07.2001, Uppsala] R&D Projects: GA ČR GA524/00/0474 Subject RIV: EC - Immunology

  11. A dolphin peripheral blood leukocyte cDNA microarray for studies of immune function and stress reactions.

    Mancia, Annalaura; Lundqvist, Mats L; Romano, Tracy A; Peden-Adams, Margie M; Fair, Patricia A; Kindy, Mark S; Ellis, Blake C; Gattoni-Celli, Sebastiano; McKillen, David J; Trent, Harold F; Chen, Yian Ann; Almeida, Jonas S; Gross, Paul S; Chapman, Robert W; Warr, Gregory W


    A microarray focused on stress response and immune function genes of the bottlenosed dolphin has been developed. Random expressed sequence tags (ESTs) were isolated and sequenced from two dolphin peripheral blood leukocyte (PBL) cDNA libraries biased towards T- and B-cell gene expression by stimulation with IL-2 and LPS, respectively. A total of 2784 clones were sequenced and contig analysis yielded 1343 unigenes (archived and annotated at ). In addition, 52 dolphin genes known to be important in innate and adaptive immune function and stress responses of terrestrial mammals were specifically targeted, cloned and added to the unigene collection. The set of dolphin sequences printed on a cDNA microarray comprised the 1343 unigenes, the 52 targeted genes and 2305 randomly selected (but unsequenced) EST clones. This set was printed in duplicate spots, side by side, and in two replicates per slide, such that the total number of features per microarray slide was 19,200, including controls. The dolphin arrays were validated and transcriptomic profiles were generated using PBL from a wild dolphin, a captive dolphin and dolphin skin cells. The results demonstrate that the array is a reproducible and informative tool for assessing differential gene expression in dolphin PBL and in other tissues. PMID:17084893

  12. Oxidative DNA damage of peripheral blood polymorphonuclear leukocytes, selectively induced by chronic arsenic exposure, is associated with extent of arsenic-related skin lesions

    There is increasing evidence that oxidative stress is an important risk factor for arsenic-related diseases. Peripheral blood leukocytes constitute an important defense against microorganisms or pathogens, while the research on the impact of chronic arsenic exposure on peripheral blood leukocytes is much more limited, especially at low level arsenic exposure. The purpose of the present study was to explore whether chronic arsenic exposure affects oxidative stress of peripheral blood leukocytes and possible linkages between oxidative stress and arsenic-induced skin lesions. 75 male inhabitants recruited from an As-endemic region of China were investigated in the present study. The classification of arsenicosis was based on the degree of skin lesions. Arsenic levels were measured in drinking water and urine by Atomic Fluorescence Spectroscopy. Urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) was tested by Enzyme-Linked Immunosorbent Assay. 8-OHdG of peripheral blood leukocytes was evaluated using immunocytochemical staining. 8-OHdG-positive reactions were only present in polymorphonuclear leukocytes (PMNs), but not in monocytes (MNs). The 8-OHdG staining of PMN cytoplasm was observed in all investigated populations, while the 8-OHdG staining of PMN nuclei was frequently found along with the elevated amounts of cell debris in individuals with skin lesion. Urinary arsenic levels were increased in the severe skin lesion group compared with the normal group. No relationship was observed between drinking water arsenic or urine 8-OHdG and the degree of skin lesions. These findings indicated that the target and persistent oxidative stress in peripheral blood PMNs may be employed as a sensitive biomarker directly to assess adverse health effects caused by chronic exposure to lower levels of arsenic. -- Highlights: ► Male inhabitants were investigated from an As-endemic region of China. ► 8-OHdG-positive reactions were only present in polymorphonuclear leukocytes (PMNs).

  13. Oxidative DNA damage of peripheral blood polymorphonuclear leukocytes, selectively induced by chronic arsenic exposure, is associated with extent of arsenic-related skin lesions

    Pei, Qiuling, E-mail: [Department of Toxicology, Public Health College, Shanxi Medical University, No 56 Xin Jian Nan Lu, Taiyuan (030001) (China); Ma, Ning [Faculty of Health Science, Suzuka University of Medical Science, Suzuka, 510-0293 (Japan); Zhang, Jing; Xu, Wenchao; Li, Yong; Ma, Zhifeng; Li, Yunyun; Tian, Fengjie; Zhang, Wenping [Department of Toxicology, Public Health College, Shanxi Medical University, No 56 Xin Jian Nan Lu, Taiyuan (030001) (China); Mu, Jinjun [The Second Hospital, Shanxi Medical University, Taiyuan (030001) (China); Li, Yuanfei [The First Hospital, Shanxi Medical University, Taiyuan (030001) (China); Wang, Dongxing; Liu, Haifang; Yang, Mimi; Ma, Caifeng; Yun, Fen [Department of Toxicology, Public Health College, Shanxi Medical University, No 56 Xin Jian Nan Lu, Taiyuan (030001) (China)


    There is increasing evidence that oxidative stress is an important risk factor for arsenic-related diseases. Peripheral blood leukocytes constitute an important defense against microorganisms or pathogens, while the research on the impact of chronic arsenic exposure on peripheral blood leukocytes is much more limited, especially at low level arsenic exposure. The purpose of the present study was to explore whether chronic arsenic exposure affects oxidative stress of peripheral blood leukocytes and possible linkages between oxidative stress and arsenic-induced skin lesions. 75 male inhabitants recruited from an As-endemic region of China were investigated in the present study. The classification of arsenicosis was based on the degree of skin lesions. Arsenic levels were measured in drinking water and urine by Atomic Fluorescence Spectroscopy. Urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) was tested by Enzyme-Linked Immunosorbent Assay. 8-OHdG of peripheral blood leukocytes was evaluated using immunocytochemical staining. 8-OHdG-positive reactions were only present in polymorphonuclear leukocytes (PMNs), but not in monocytes (MNs). The 8-OHdG staining of PMN cytoplasm was observed in all investigated populations, while the 8-OHdG staining of PMN nuclei was frequently found along with the elevated amounts of cell debris in individuals with skin lesion. Urinary arsenic levels were increased in the severe skin lesion group compared with the normal group. No relationship was observed between drinking water arsenic or urine 8-OHdG and the degree of skin lesions. These findings indicated that the target and persistent oxidative stress in peripheral blood PMNs may be employed as a sensitive biomarker directly to assess adverse health effects caused by chronic exposure to lower levels of arsenic. -- Highlights: ► Male inhabitants were investigated from an As-endemic region of China. ► 8-OHdG-positive reactions were only present in polymorphonuclear leukocytes (PMNs).

  14. Age-associated DNA methylation changes in immune genes, histone modifiers and chromatin remodeling factors within 5 years after birth in human blood leukocytes

    Acevedo, Nathalie; Reinius, Lovisa E; Vitezic, Morana;


    BACKGROUND: Age-related changes in DNA methylation occurring in blood leukocytes during early childhood may reflect epigenetic maturation. We hypothesized that some of these changes involve gene networks of critical relevance in leukocyte biology and conducted a prospective study to elucidate the...... dynamics of DNA methylation. Serial blood samples were collected at 3, 6, 12, 24, 36, 48 and 60 months after birth in ten healthy girls born in Finland and participating in the Type 1 Diabetes Prediction and Prevention Study. DNA methylation was measured using the HumanMethylation450 BeadChip. RESULTS......: After filtering for the presence of polymorphisms and cell-lineage-specific signatures, 794 CpG sites showed significant DNA methylation differences as a function of age in all children (41.6% age-methylated and 58.4% age-demethylated, Bonferroni-corrected P value <0.01). Age-methylated CpGs were more...

  15. Peripheral Blood Leukocyte Production of BDNF following Mitogen Stimulation in Early Onset and Regressive Autism

    Amanda Enstrom


    Full Text Available Brain-derived neurotrophic factor (BDNF is critical for neuronal differentiation and synaptic development. BDNF is also implicated in the development of psychological disorders including depression, bipolar disorder and schizophrenia. Previously, elevated BDNF levels were observed in neonatal blood samples from infants who were later diagnosed with autism when compared with children who developed normally, suggesting that BDNF may be involved in the development of autism. BDNF is produced by activated brain microglial cells, a cellular phenotype that shares several features with peripheral macrophages, suggesting an important role for the immune system in BDNF production. We hypothesized that under mitogenic stimulation, peripheral blood mononuclear cells obtained from children with autism may have altered BDNF production compared with age-matched typically developing control subjects. In addition, we examined the differences between the production of BDNF in classic/early-onset autism and children who had a regressive form of autism. We show here that plasma levels of BDNF levels are increased in children with autism, especially in early onset autism subjects. Furthermore, under mitogenic stimulation with PHA and LPS, BDNF production is significantly increased in children with autism compared with typically developing subjects. However, stimulation with tetanus toxoid results in a decreased response in children with autism. This data suggest that immune cell-derived production of BDNF could be an important source for the increased BDNF that is detected in some subjects with autism. As a neurotrophic factor produced by immune cells, BDNF could help elucidate the role of the immune system in neurodevelopment and neuronal maintenance, which may be dysregulated in autism.

  16. Identification of an additional class of C3-binding membrane proteins of human peripheral blood leukocytes and cell lines.

    Cole, J L; Housley, G A; Dykman, T R; MacDermott, R P; Atkinson, J P


    Proteins binding the third component of complement (C3) were isolated by affinity chromatography from surface-labeled solubilized membranes of human peripheral blood cells and cell lines. The isolated molecules were subjected to NaDodSO4/PAGE, and autoradiographs of these gels indicated that C3-binding proteins could be divided into three groups based on Mr: (i) gp200, an approximately 200,000 Mr molecule previously identified as the C3b/C4b receptor or CR1; (ii) gp140, an approximately 140,000 Mr molecule previously identified as the C3d receptor or CR2; and (iii) gp45-70, a heretofore unrecognized group of 45,000-70,000 Mr C3-binding molecules. The cell distribution, Mr, antigenic cross-reactivity, and specificity of gp45-70 were examined. Erythrocytes have no detectable gp45-70, but all leukocyte populations examined possess this group of molecules. On neutrophils and mononuclear phagocytes, CR1 is the predominant C3-binding glycoprotein, but gp45-70 is present on both cell populations and on macrophage and neutrophil cell lines. B plus null cells, chronic lymphocytic leukemia cells, and an Epstein-Barr virus-transformed B-cell line possess CR1, CR2, and gp45-70. On T cells and T-cell lines gp45-70 is the predominant or, in some cases, the only C3-binding protein isolated. gp45-70 is structurally characterized as a broad band or doublet with a mean Mr that is slightly different for each cell population. gp45-70 binds iC3, C3b, and C4b, but not C3d, indicating that the binding region is probably within the C3c portion of C3b. A polyclonal antibody to CR1 and monoclonal antibodies to CR1 and CR2 do not immunoprecipitate gp45-70. While gp45-70 has not been previously characterized on human cells, a C3b-binding glycoprotein of similar Mr is present on rabbit alveolar macrophages. We conclude that gp45-70 is an additional group of membrane proteins present on human leukocytes that possess ligand-binding activity for C3b. PMID:3871945

  17. Hesperidin displays relevant role in the nutrigenomic effect of orange juice on blood leukocytes in human volunteers: a randomized controlled cross-over study

    Deval, Christiane; Dubray, Claude; Mazur, Andrzej; Morand, Christine


    Background We previously showed, in healthy, middle-aged, moderately overweight men, that orange juice decreases diastolic blood pressure and significantly improves postprandial microvascular endothelial reactivity and that hesperidin could be causally linked to the observed beneficial effect of orange juice. The objective was to determine the effect of chronic consumption of orange juice on the gene expression profile of leukocytes in healthy volunteers and to assess to what extent hesperidi...

  18. Interference with human immunodeficiency virus (HIV) replication by CD8+ T cells in peripheral blood leukocytes of asymptomatic HIV carriers in vitro.

    Kannagi, M; Masuda, T; Hattori, T; Kanoh, T.; Nasu, K; Yamamoto, N; Harada, S


    A long asymptomatic period is one of the characteristics of human immunodeficiency virus (HIV) infection, despite its fatal consequences. Antiviral defense in HIV-infected individuals controls viral replication during this period. In the present study, we demonstrate that peripheral blood leukocytes (PBL) of asymptomatic HIV-1 carriers, following exogenous HIV-1 infection in vitro, do not support viral replication. These cells do not produce detectable amounts of reverse transcriptase or accu...

  19. Imaging with FDG labelled leukocytes: is it clinically useful?

    In vivo and in vitro labeled leukocytes have been shown to be very effective in detecting different infectious and inflammatory conditions. The model of labeled leukocyte imaging is based on the powerful mechanisms of chemotaxis exerted on activated leukocytes by chemo-attractants. The avidity of inflammatory cells for fluorodeoxyglucose (FDG) has led to the concept of labeling leukocytes with [18 F]FDG ex vivo. This concept combines cell-bound radionuclide trafficking from the blood pool compartment to the lesion with the high resolution of positron emission tomography (PET) imaging. The further benefits of having a correlated anatomical map by implementing the acquisition on a hybrid PET/computed tomography (CT) device are obvious. The feasibility and the potential value of leukocyte PET(/CT) imaging in infection have been demonstrated. The available data suggest a high accuracy of the method. Still, leukocyte PET/CT should not be considered as the endpoint of infection imaging, since it only meets a part of the criteria of the ideal infection imaging agent. However, the common clinical need for specific detection of infection with anatomical precision, the availability of the components necessary for performing leukocyte PET/CT, their lack of toxicity or adverse effects and the absence of more superior tracers on the commercial market make it worthwhile to further investigate leukocyte PET/CT imaging in larger prospective series. The advantages of leukocyte PET/CT over the more conventional nuclear medicine and radiological methods makes this imaging tool likely to be useful in certain subsets of infected patients.

  20. Study on serum TNF-α level, B-cell count and T-cell subsets distribution in peripheral blood in patients with rheumatoid arthritis

    Objective: To study the changes of serum TNF-α levels, B-cell count and T-cell subsets distribution in peripheral blood in patients with rheumatoid arthritis. Methods: Serum TNF-α levels (with RIA), B cell as well as T cell subsets distribution type (with monoclonal antibody technique) were examined in 37 patients with rheumatoid arthritis and 30 controls. Results Serum TNF-α levels and B lymphocytes count were significantly higher in the patients than those in controls (P3, CD4 and CD4/CD8 were obviously lower (P<0.01). Conclusion: Rheumatoid arthritis is an autoimmune disease with abnormal immunoregulation. (authors)

  1. Differential innate immune responses of bovine peripheral blood leukocytes to Salmonella enterica serovars Dublin, Typhimurium, and Enteritidis.

    Pan, Deng; Rostagno, Marcos H; Ebner, Paul D; Eicher, Susan D


    The majority of Salmonella serovars cause no clinical disease in cattle, while some are associated with severe disease. The objective of the current study was to determine the innate immune responses of bovine peripheral blood leukocytes exposed to Salmonella enterica serovar Dublin (bovine-specific), Salmonella typhimurium (murine adapted, but zoonotic), and Salmonella enteritidis (poultry host-adapted) in 3-week-old calves. All Salmonella exposures increased cell surface CD14 and CD18 regardless of serovar. The greatest CD14 marker mean fluorescence was in monocytes and the greatest mean fluorescent of the marker mean was in neutrophils. Phagocytosis increased with all serovars, but was not different among them. Neutrophils had the greatest marker mean fluorescence for phagocytosis, with all serovars being equal. Oxidative burst increased in all serovars compared to control cells, but were not different among the serovars. Neutrophils and monocytes were similar in the oxidative burst, with limited oxidative burst detected in the primarily lymphocyte population. mRNA expression of TNF-α, IL-8, and IL-12, increased above the control cells whereas none of these serovars affected mRNA expression of TLR4. TNF-α was greatest in S. enterica and S. typhimurium, compared to Salmonella dublin. In contrast, IL-8 was expressed more in S. dublin than S. typhiurium, with S. Enteriditus intermediary. These results show while cell surface markers, phagocytosis, and oxidative burst were largely unaffected by serovar, cytokine and chemokine expression differed among the Salmonella serovars. It appears that internal responses of the cells differ, rather than cell recognition, creating pathogenicity differences among of the serovars, even in the neonate with developing immunity. PMID:25847354

  2. Activity of superior interferon α against HIV-1 in severe combined immunodeficient mice reconstituted with human peripheral blood leukocytes

    ZHANG Wei; TONG Xiao; Tadashi Nakasone; YUE Xue-tian; Naoki Yamamoto; LIU Xin-yuan; YANG Rong-ge


    Background Interferon (IFN) can inhibit human immunodeficiency virus type 1 (HIV-1) replication in vitro and in clinic.However, IFN therapy for HIV infection was limited by its moderate antiviral efficacy and its frequent adverse effects. In the present study we evaluated the anti-HIV efficacy of a novel synthesized superior interferon α (slFNα).Methods We performed in vitro experiments with HIV-1 IIB infected MT4 cells, and evaluated in vivo anti-HIV efficacy of slFNα in severe combined immunodeficient (SClD) mice reconstituted with human peripheral blood leukocytes (hu-PBL-SClD mice).Results We found that the 50% effective concentrations (EC5o) of slFNα against the replication of HIV-1 in MT4 cells was 0.06 ng/ml, representing stronger antiviral activity than interferon-α in vitro. In the hu-PBL-SCID mice, a dose-dependent protection pattern was observed: with 0.45 μg and 1.35 μg slFNα daily treatments, parts of SCID mice were protected from HIV infection, whereas 2.25 μg sIFNα daily treatments resulted in a terminally complete protection.Conclusions slFNα shows good anti-HIV activity both in vitro and in SCID mice, may be a promising anti-HIV agent deserving clinical investigation, especially considering the potential of IFN-α to inhibit HIV replication in patients infected with drug-resistant variants or co-infected with hepatitis C virus (HCV).

  3. Molecular features of the complementarity determining region 3 motif of the T cell population and subsets in the blood of patients with chronic severe hepatitis B

    Yang Jiezuan


    Full Text Available Abstract Background T cell receptor (TCR reflects the status and function of T cells. We previously developed a gene melting spectral pattern (GMSP assay, which rapidly detects clonal expansion of the T cell receptor β variable gene (TCRBV in patients with HBV by using quantitative real-time reverse transcription PCR (qRT-PCR with DNA melting curve analysis. However, the molecular profiles of TCRBV in peripheral blood mononuclear cells (PBMCs and CD8+, CD8- cell subsets from chronic severe hepatitis B (CSHB patients have not been well described. Methods Human PBMCs were separated and sorted into CD8+ and CD8- cell subsets using density gradient centrifugation and magnetic activated cell sorting (MACS. The molecular features of the TCRBV CDR3 motif were determined using GMSP analysis; the TCRBV families were cloned and sequenced when the GMSP profile showed a single-peak, indicative of a monoclonal population. Results The number of skewed TCRBV in the CD8+ cell subset was significantly higher than that of the CD8- cell subset as assessed by GMSP analysis. The TCRBV11 and BV7 were expressed more frequently than other members of TCRBV family in PBMCs and CD8+, CD8- subsets. Also the relatively conserved amino acid motifs were detected in the TCRBV22, BV18 and BV11 CDR3 in PBMCs among patients with CSHB. Conclusions The molecular features of the TCRBV CDR3 were markedly different among PBMCs and CD8+, CD8- cell subsets derived from CSHB patients. Analysis of the TCRBV expression in the CD8+ subset was more accurate in assessing the status and function of circulating T cells. The expression of TCRBV11, BV7 and the relatively conserved CDR3 amino acid motifs could also help to predict and treat patients with CSHB.

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

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


    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

  5. Siglec-1-positive plasmacytoid dendritic cells (pDCs) in human peripheral blood: A semi-mature and myeloid-like subset imbalanced during protective and autoimmune responses.

    Wilhelm, Theresa R; Taddeo, Adriano; Winter, Oliver; Schulz, Axel Ronald; Mälzer, Julia-Nora; Domingo, Cristina; Biesen, Robert; Alexander, Tobias; Thiel, Andreas; Radbruch, Andreas; Hiepe, Falk; Gerl, Velia


    Plasmacytoid dendritic cells (pDCs) play a central role in the pathogenesis of systemic lupus erythematosus (SLE) as IFN-α producers and promoters of T-cell activation or tolerance. Here, we demonstrated by flow-cytometry and confocal microscopy that Siglec-1, a molecule involved in the regulation of adaptive immunoresponses, is expressed in a subset of semi-mature, myeloid-like pDCs in human blood. These pDCs express lower BDCA-2 and CD123 and higher HLA-DR and CD11c than Siglec-1-negative pDCs and do not produce IFN-α via TLR7/TLR9 engagement. In vitro, Siglec-1 expression was induced in Siglec-1-negative pDCs by influenza virus. Proportions of Siglec-1-positive/Siglec-1-negative pDCs were higher in SLE than in healthy controls and correlated with disease activity. Healthy donors immunized with yellow fever vaccine YFV-17D displayed different kinetics of the two pDC subsets during protective immune response. PDCs can be subdivided into two subsets according to Siglec-1 expression. These subsets may play specific roles in (auto)immune responses. PMID:26674280

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

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


    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.

  7. [A new separation protocol (DRBCP-F) for automated blood component donation with the MCS 3p cell separator for collection of leukocyte depleted erythrocyte concentrates and plasma].

    Zeiler, T; Kretschmer, V


    Previously published studies on automated blood component donation with the MCS 3p cell separator proved fairly good quality of the collected red blood cells (RBC) and fresh frozen plasma (FFP), with the disadvantage of a low hematocrit of the filtered RBC and a high platelet contamination of the FFP (RBCP-F protocol.) The DRBCP-F protocol was designed to eliminate the above-mentioned disadvantages and to provide 1 unit of leuko-depleted (filtered) RBC, 2 units of FFP, and additionally 1 platelet concentrate (PC) from the buffy coat. Twenty automated blood component collections (2 cycles, Latham bowl at 5,500 rpm, 230 ml isotonic saline for volume balance, PAGGS-M as additive solution) were performed. The RBC were filtered in a closed system after storage at 4 degrees C for 24 h. Blood cell counts and biochemical parameters of the RBC were determined initially and after 49 days. PC were separated from buffy coat after a soft spin. The volume of the RBC amounted to 293 +/- 12 ml (mean +/- SD) with a hematocrit of 0.61 +/- 0.05 l/l. Residual leukocytes after filtration were found to be 0.04 x 10(6) +/- 0.06 per unit. After storage, the following data were obtained: hemolysis 0.38%, ATP 2.1 +/- 0.4 mumol/g Hb, 2,3-diphosphoglycerate (2,3-DPG) 1.4 +/- 0.3 mumol/g Hb, ph 6.3 +/- 0.1, potassium 6.4 mmol per unit, and LDH in the supernatant was 219 U/l. None of the RBC showed bacterial growth after 49 days. The volume of the collected FFP was 398 +/- 32 ml, with 3.4 +/- 3.5 x 10(3) residual platelets and 5 +/- 12 leukocytes per microliter. Platelet concentrates contained 90.2 +/- 32 x 10(9) platelets in 88 +/- 14 ml plasma. Automated blood donation with the DRBCP-F protocol provided RBC with very low residual leukocyte counts, adequate hematocrit and good metabolic status up to 49 days, and FFP with low platelet contamination. The platelet concentrates were even superior to those prepared from whole blood using the buffy coat method. The storable leuko-depleted RBC are

  8. Hesperidin displays relevant role in the nutrigenomic effect of orange juice on blood leukocytes in human volunteers: a randomized controlled cross-over study.

    Dragan Milenkovic

    Full Text Available BACKGROUND: We previously showed, in healthy, middle-aged, moderately overweight men, that orange juice decreases diastolic blood pressure and significantly improves postprandial microvascular endothelial reactivity and that hesperidin could be causally linked to the observed beneficial effect of orange juice. The objective was to determine the effect of chronic consumption of orange juice on the gene expression profile of leukocytes in healthy volunteers and to assess to what extent hesperidin is involved in the effect of orange juice. METHODOLOGY/PRINCIPAL FINDINGS: Volunteers were included in a randomized, controlled, crossover study. Throughout three 4-week periods, volunteers consumed daily: 500 ml orange juice, 500 ml control drink plus hesperidin or 500 ml control drink and placebo. Blood samplings were performed on 10 overnight-fasted subjects after the 4-week treatment period. Global gene expression profiles were determined using human whole genome cDNA microarrays. Both orange juice and hesperidin consumption significantly affected leukocyte gene expression. Orange juice consumption induced changes in expression of, 3,422 genes, while hesperidin intake modulated the expression of 1,819 genes. Between the orange juice and hesperidin consumption groups, 1,582 regulated genes were in common. Many of these genes are implicated in chemotaxis, adhesion, infiltration and lipid transport, which is suggestive of lower recruitment and infiltration of circulating cells to vascular wall and lower lipid accumulation. CONCLUSIONS: This study shows that regular consumption of orange juice for 4 weeks alters leukocyte gene expression to an anti-inflammatory and anti-atherogenic profile, and hesperidin displays a relevant role in the genomic effect of this beverage. TRIAL REGISTRATION: NCT 00983086.

  9. Flow cytometric characterizations of leukocyte subpopulations in the peripheral blood of northern pig-tailed macaques (Macaca leonina)

    Zheng, Hong-Yi; ZHANG, Ming-Xu; ZHANG, Lin-Tao; Zhang, Xiao-Liang; Pang, Wei; LYU, Long-Bao; Zheng, Yong-Tang


    Pig-tailed macaques (Macaca nemistrina group) have been extensively used as non-human primate animal models for various human diseases in recent years, notably for AIDS research due to their sensitivity to HIV-1. Northern pig-tailed macaques (M. leonina) are distributed in China and other surrounding Southeast Asia countries. Although northern pig-tailed macaques have been bred on a large scale as experimental animals since 2012, the reference value of normal levels of leukocytes is not avail...

  10. High affinity capture and concentration of quinacrine in polymorphonuclear neutrophils via vacuolar ATPase-mediated ion trapping: Comparison with other peripheral blood leukocytes and implications for the distribution of cationic drugs

    Roy, Caroline; Gagné, Valérie; Fernandes, Maria J.G.; Marceau, François, E-mail:


    Many cationic drugs are concentrated in acidic cell compartments due to low retro-diffusion of the protonated molecule (ion trapping), with an ensuing vacuolar and autophagic cytopathology. In solid tissues, there is evidence that phagocytic cells, e.g., histiocytes, preferentially concentrate cationic drugs. We hypothesized that peripheral blood leukocytes could differentially take up a fluorescent model cation, quinacrine, depending on their phagocytic competence. Quinacrine transport parameters were determined in purified or total leukocyte suspensions at 37 °C. Purified polymorphonuclear leukocytes (PMNLs, essentially neutrophils) exhibited a quinacrine uptake velocity inferior to that of lymphocytes, but a consistently higher affinity (apparent K{sub M} 1.1 vs. 6.3 μM, respectively). However, the vacuolar (V)-ATPase inhibitor bafilomycin A1 prevented quinacrine transport or initiated its release in either cell type. PMNLs capture most of the quinacrine added at low concentrations to fresh peripheral blood leukocytes compared with lymphocytes and monocytes (cytofluorometry). Accumulation of the autophagy marker LC3-II occurred rapidly and at low drug concentrations in quinacrine-treated PMNLs (significant at ≥ 2.5 μM, ≥ 2 h). Lymphocytes contained more LAMP1 than PMNLs, suggesting that the mass of lysosomes and late endosomes is a determinant of quinacrine uptake V{sub max}. PMNLs, however, exhibited the highest capacity for pinocytosis (uptake of fluorescent dextran into endosomes). The selectivity of quinacrine distribution in peripheral blood leukocytes may be determined by the collaboration of a non-concentrating plasma membrane transport mechanism, tentatively identified as pinocytosis in PMNLs, with V-ATPase-mediated concentration. Intracellular reservoirs of cationic drugs are a potential source of toxicity (e.g., loss of lysosomal function in phagocytes). - Highlights: • Quinacrine is concentrated in acidic organelles via V-ATPase-mediated ion

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

    Donskov, F; Bennedsgaard, K M; Von Der Maase, H; Marcussen, N; Fisker, R; Jensen, J J; Naredi, P; Hokland, M


    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......-six patients with metastatic renal cell carcinoma were treated with low dose s.c. interleukin-2, interferon-alpha and histamine. A total of 250 blood samples and 62 core needle biopsies from 23 and 19 of these patients, respectively, were analysed. After 2 weeks of treatment, a significant positive correlation...... 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...

  12. 滤除白细胞异体血对围术期患儿细胞免疫功能的影响%Effects of leukocyte-depleted allogeneic blood transfusion on perioperative cellular immunity in children

    邢准; 王秋实; 刁艳妮


    Objective To evaluate the effects of leukocyte-depleted allogeneic blood transfusion on perioperative cellular immunity in children.Methods Three hundred and fifty-nine ASA Ⅰ or Ⅱ children (aged 3 months-14 years and weighing 5-74 kg) requiring allogeneic blood transfusion during operation were randomly divided into two groups:163 children receiving normal allogeneic blood transfusion (control group,group C) and 196 children receiving leukocyte-depleted allogeneic blood transfusion (group D).Blood samples were collected from the peripheral vein before blood transfusion,and 2 and 6 days after blood transfusion for determination of the levels of CD3+,CD4+,CD8 +,and CD56+ by flow cytometry.CD4+ /CD8+ ratio was calculated.The volume of allogeneic blood transfusion during operation,the duration of operation,postoperative drainage,antibiotic administration,hospital stay and the incidence of postoperative infection were recorded.Rssults The levels of CD3+,CD4+,CD56+ and CD4+/CD8+ ratio significantly increased at 6 days after blood transfusion while the duration of postoperative drainage,postoperative antibiotic administration,hospital stay and the incidence of postoperative infection significantly decreased in group D compared with group C.Conclusion Leukocyte-depleted allogeneic blood transfusion is helpful in improving the postoperative cellular immunity in children.

  13. Determination of the effects of extremely low frequency electromagnetic fields on the percentages of peripheral blood leukocytes and histology of lymphoid organs of the mouse

    Objective was to determine the effects of very weak, extremely low frequency (50Hz) electromagnetic field (ELF-EMF) on the relative spleen weight, lymphoid organ histology, peripheral blood leukocyte and alpha-naphthyl acetate esterase positive (ANAE-positive) lymphocyte percentages of mouse. The study was carried out in the Scientific Research and Application Center of Selcuk University, Konya, Turkey in 2005. A total of 120 Swiss albino mice were divided into 6 groups (20 in each group). The experimental animals were exposed to 1, 2, 3, 4 and 5 uT flux intensities (rms) of EMF at 50 Hz for 40 days. In the exposure groups with 20 animals, the body weight (BW) increased gradually in higher field intensities and reached at peak level in the 4 uT and then slightly decreased. The relative spleen weight (% of BW) was not affected. The ELF-EMF treatment did not cause any significant change in lymphocyte, monocyte and ANA-positive lymphocyte ratios, whereas percentages of neutropholis and basophiles changed non-linearly. Any change in the lymphoid organ histology which attributable to the field effect, was not observed in the exposure groups. Extremely frequency-EMF exposure with the flux intensities between 1-5 uT for 40 days did not cause any effect on the relative spleen weight, lymphoid organ histology, leukocyte and ANAE-positive lymphocyte percentages of the mouse. (author)

  14. Dynamic expression of leukocyte innate immune genes in whole blood from horses with lipopolysaccharide-induced acute systemic inflammation

    Vinther, Anne Mette L.; Skovgaard, Kerstin; Heegaard, Peter M. H.;


    Background: In horses, insights into the innate immune processes in acute systemic inflammation are limited even though these processes may be highly important for future diagnostic and therapeutic advances in high-mortality disease conditions as the systemic inflammatory response syndrome (SIRS...... were compared with baseline levels. Results: Systemic inflammation was confirmed by the presence of clinical and hematological changes which were consistent with SIRS. The clinical response to LPS was transient and brief as all horses except one showed unaltered general demeanor after 24 h. Twenty......-two leukocyte genes were significantly regulated at at least one time point during the experimental period. By close inspection of the temporal responses the dynamic changes in mRNA abundance revealed a very rapid onset of both pro-and anti-inflammatory mediators and a substantial variation in both expression...

  15. Analysis of chromosomal aberrations of leukocytes of peripheric blood in patients with depression of different genesis hematopoiesis

    Cytogenic investigations were carried on to reveal the increase in the aberration cell content as compared to the spontaneous level in 30 patients. The least quantity of the cells were in patients exposed to radiotherapy (6-14%), the greatest - in patients subjected to the combined treatment (radio- and chemotherapies). All the patients had structural changes in chromosomes and the primary type of aberrations was a chromosomal one. The group of patients subjected to chemotherapy before leukopenia had aberrations of the chromatid type (16.4%). It is concluded that leukopenias result from structural and quantitative changes in the chromosomal apparatus of leukocytes in patients under the effect of mutagenous factors on the organism

  16. The comparison of ordered subset expectation maximization and filtered back projection technique for RBC blood pool SPECT in detection of liver hemangioma

    Jeon, Tae Joo; Kim, Hee Joung; Bong, Jung Kyun; Lee, Jong Doo [College of Medicine, Yonsei Univ., Seoul (Korea, Republic of)


    Odered subset expectation maximization (OSEM) is a new iterative reconstruction technique for tomographic images that can reduce the reconstruction time comparing with conventional iteration method. We adopted this method of RBC blood pool SPECT and tried to validate the usefulness of OSEM in detection of liver hemangioma comparing with filtered back projection (FBP). A 64 projection SPECT study was acquired over 360 .deg. C by dual-head cameras after the injection of 750MBq of {sup 99m}Tc-RBC. OSEM was performed with various condition of subset (1,2,4,8,16 and 32) and iteration number (1,2,4,8 and 16) to obtain the best set for lesion detection. OSEM underwent in 17 lesions of 15 patients with liver hemangioma and compared with FBP images. Two nuclear medicine physicians reviewed these results independently. Best set for images was 4 iteration and 16 subset. In general, OSEM revealed more homogeneous images than FBP. Eighty-eight percent (15/17) of OSEM images were superior or equal to FBP for anatomic resolution. According to the blind review of images 70.5% (12/17) of OSEM was better in contrast (4/17), anatomic detail (4/17) and both (2/17). Two small lesions were detected by OSEM only and another 2 small lesions were failed to depict in both methods. Remaining 3 lesions revealed no difference in image quality. OSEM can provide better image quality as well as better results in detection of liver hemangioma than conventional FBP technique.

  17. Modification of leukocyte adherence inhibition (LAI) assay with 51Cr-labelled leukocytes

    Leukocyte adherence inhibition was measured by a new modified radioisotopic technique. Peripheral blood leukocytes were isolated and labelled with 51Cr. These leukocytes were incubated with medium or buffer alone and with medium or buffer containing tumor antigen or gluten. The glass surface for the adherence was prepared carefully. In all samples the adherence of leukocytes occured under the same conditions. The results of the LAI assay with gluten and stomach cancer antigen are reported and discussed. (author)

  18. Flow-cytometric analysis of changes in lymphocyte subsets in the blood of cancer patients during radiotherapy, 1

    In this study, we report a clinical application of a rapid flow-cytometric immunofluorescence method for studying OKT11+-lymphocytes in whole blood by using monoclonal antibody OKT11. Changes in the relative percentages and absolute counts of OKT11+-lymphocytes during radiotherapy of cancer patients were evaluated by this method. In a study of 151 samples from 58 cases, the elevation of OKT11+-lymphocyte level (>=90%) was observed in 38 samples from 21 patients. Many cases of these responded well to radiotherapy. In contrast, the depression of OKT11+-lymphocyte level (+-lymphocytes were consistent during radiotherapy, but absolute counts of OKT11+-lymphocytes were remarkably decreased during radiotherapy. In a study of 40 samples, the correlation coefficient between the manual E-rosette count and the flow immunofluorescence measurement was 0.8914. In a study of 132 samples, that between the absolute lymphocyte count and the OKT11+-lymphocyte count was 0.9673. (author)

  19. Monoclonal antibody to a subset of human monocytes found only in the peripheral blood and inflammatory tissues

    Zwadlo, G.; Schlegel, R.; Sorg, C.


    A monoclonal antibody is described that was generated by immunizing mice with cultured human blood monocytes. The antibody (27E10) belongs to the IgG1 subclass and detects a surface antigen at M/sub r/ 17,000 that is found on 20% of peripheral blood monocytes. The antigen is increasingly expressed upon culture of monocytes, reaching a maximum between days 2 and 3. Stimulation of monocytes with interferon-..gamma.. (IFN-..gamma..), 12-O-tetradecanoyl-phorbol-13-acetate (TPA), and lipopolysaccharide (LPS) Ylalanine (fMLP) increased the 27E10 antigen density. The amount of 27E10-positive cells is not or is only weakly affected. The antigen is absent from platelets, lymphotyces, and all tested human cell lines, yet it cross-reacts with 15% of freshly isolated granulocytes. By using the indirect immunoperoxidase technique, the antibody is found to be negative on cryostat sections of normal human tissue (skin, lung, and colon) and positive on only a few monocyte-like cells in liver and on part of the cells of the splenic red pulp. In inflammatory tissue, however, the antibody is positive on monocytes/macrophages and sometimes on endothelial cells and epidermal cells, depending on the stage and type of inflammation, e.g., BCG ranulomas are negative, whereas psoriasis vulgaris, atopic dermatitis, erythrodermia, pressure urticaria, and periodontitis contain positively staining cells. In contact eczemas at different times after elicitation (6 hr, 24 hr, and 72 hr), the 27E10 antigen is seen first after 24 hr on a few infiltrating monocytes/macrophages, which increase in numbers after 72 hr.

  20. Pathologic and Protective Roles for Microglial Subsets and Bone Marrow- and Blood-Derived Myeloid Cells in Central Nervous System Inflammation

    Wlodarczyk, Agnieszka; Cédile, Oriane; Jensen, Kirstine Nolling;


    Inflammation is a series of processes designed for eventual clearance of pathogens and repair of damaged tissue. In the context of autoimmune recognition, inflammatory processes are usually considered to be pathological. This is also true for inflammatory responses in the central nervous system...... (CNS). However, as in other tissues, neuroinflammation can have beneficial as well as pathological outcomes. The complex role of encephalitogenic T cells in multiple sclerosis and its animal model experimental autoimmune encephalomyelitis (EAE) may derive from heterogeneity of the myeloid cells with...... three populations of myeloid cells: CD11c(+) microglia, CD11c(-) microglia, and CD11c(+) blood-derived cells in terms of their pathological versus protective functions in the CNS of mice with EAE. Our data show that CNS-resident microglia include functionally distinct subsets that can be distinguished...

  1. Longitudinal changes of peripheral blood DC subsets and regulatory T cells in Chinese chronic HIV-1-infected patients during antiretroviral therapy.

    Mei Zhang

    Full Text Available It has been emphasized that chronic generalized immune dysfunction is the leading event in the pathogenesis of HIV infection, in which the contribution of dendritic cells (DCs and regulatory T cells (Tregs should not be underestimated. In current study, we assessed the longitudinal changes of peripheral blood DC subsets and Tregs in chronically asymptomatic treatment-naive HIV-1-infected patients during 60 weeks of antiretroviral therapy (ART, and compared with those in healthy controls and long term non-progressors (LTNPs. Blood samples were collected at week 0, 4, 12, 24, 48 and 60 of treatment to measure the counts of DC subsets and Tregs by flow cytometry and IFN-a plasma levels by ELISA. The counts of myeloid dendritic cells (mDCs increased during ART, reaching similar levels to healthy controls at week 60 post ART but still lower than those of LTNPs. In HIV-1-infected patients, the mDCs counts were directly correlated with CD4 counts during ART. Changes in mDCs at week 8 were positively correlated with the changes in CD4 counts at week 60 post ART. However, the counts and function of plasmacytoid dendritic cells (pDCs remained relatively stable during ART, and similar to those in healthy controls and LTNPs. The percentage of Tregs increased before ART and normalized after ART. Importantly, we found pDCs counts were associated with percentage of Tregs during ART, which may help in understanding of the role of these cells in HIV infection.

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

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


    [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.

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  1. Perioperative transfusion of leukocyte depleted blood products in gastric cancer patients negatively influences oncologic outcome: A retrospective propensity score weighted analysis on 610 curatively resected gastric cancer patients.

    Reim, Daniel; Strobl, Andreas N; Buchner, Christian; Schirren, Rebekka; Mueller, Werner; Luppa, Peter; Ankerst, Donna Pauler; Friess, Helmut; Novotny, Alexander


    The influence of perioperative transfusion (PT) on outcome following surgery for gastric cancer (GC) remains controversial, with randomized trials lacking and observational series confounded by patient risk factors. This analysis determines the association between reception of leukocyte-depleted blood products and post-operative survival for GC.Data from 610 patients who underwent curative surgery for GC in a German tertiary care clinic from 2001 to 2013 were included. Kaplan-Meier survival curves and Cox proportional hazards regression were applied to determine the association of PT and clinical and patient risk factors for overall and relapse-free survival. Propensity score analysis was performed to adjust for observational biases in reception of PT.Higher Union International Contre le Cancer/American Joint Committee on Cancer (UICC/AJCC)-stages (P female gender (P = 0.03). In the adjusted propensity score weighted analysis, PT remained associated with an increased risk of death (hazard ratio (HR): 1.31, 95% CI: 1.01-1.69, P = 0.04).Because of the association of PT with negative influence on patient survival following resection for GC, risks from application of blood products should be weighed against the potential benefits. PMID:27442682

  2. Whole transcriptome RNA sequencing data from blood leukocytes derived from Parkinson's disease patients prior to and following deep brain stimulation treatment

    Lilach Soreq


    Full Text Available Recent evidence demonstrates the power of RNA sequencing (RNA-Seq for identifying valuable and urgently needed blood biomarkers and advancing both early and accurate detection of neurological diseases, and in particular Parkinson's disease (PD. RNA sequencing technology enables non-biased, high throughput, probe-independent inspection of expression data and high coverage and both quantification of global transcript levels as well as the detection of expressed exons and junctions given a sufficient sequencing depth (coverage. However, the analysis of sequencing data frequently presents a bottleneck. Tools for quantification of alternative splicing from sequenced libraries hardly exist at the present time, and methods that support multiple sequencing platforms are especially lacking. Here, we describe in details a whole RNA-Seq transcriptome dataset produced from PD patient's blood leukocytes. The samples were taken prior to, and following deep brain stimulation (DBS treatment while being on stimulation and following 1 h of complete electrical stimulation cessation and from healthy control volunteers. We describe in detail the methodology applied for analyzing the RNA-Seq data including differential expression of long noncoding RNAs (lncRNAs. We also provide details of the corresponding analysis of in-depth splice isoform data from junction and exon reads, with the use of the software AltAnalyze. Both the RNA-Seq raw ( and analyzed data (!Synapse:syn2805267 may be found valuable towards detection of novel blood biomarkers for PD.

  3. Phagocytic and oxidative-burst activity of blood leukocytes in rats fed a protein-free diet

    Sawosz, Ewa; Winnicka, Anna; Chwalibog, André;


    The objective of this study was to evaluate the effects of two weeks' protein deprivation on the cellular parameters of non-specific immunity in rats. Wistar rats (200-250 g) were divided into two groups (2x12) and were fed two isoenergetic (control and protein-free) diets. The phagocytic activity...... of neutrophils and monocytes, and the oxidative-burst activity of neutrophils of peripheral blood, were determined by flow cytometry after stimulation with E. coli and phorbol 12-mirystate 13-acetate. Feeding the protein-free diet for two weeks did not influence the phagocytic activity of neutrophils......, monocytes or blood morphology. However, the oxidative burst of stimulated neutrophils was increased indicating that two weeks' protein deprivation does not depress the oxygen-dependent killing mechanism in neutrophils, but may lead to the overproduction of reactive oxygen species....

  4. STAT1 signaling modulates HIV-1–induced inflammatory responses and leukocyte transmigration across the blood-brain barrier

    Chaudhuri, Anathbandhu; Yang, Bo; Gendelman, Howard E; Persidsky, Yuri; Kanmogne, Georgette D.


    The relationship among neuroinflammation, blood-brain barrier (BBB) dysfunction, and progressive HIV-1 infection as they affect the onset and development of neuroAIDS is incompletely understood. One possible link is signal transducers and activators of transcription (STATs) pathways. These respond to proinflammatory and regulatory factors and could affect neuroinflammatory responses induced from infected cells and disease-affected brain tissue. Our previous works demonstrated that HIV-1 activ...

  5. Disorders of sex development expose transcriptional autonomy of genetic sex and androgen-programmed hormonal sex in human blood leukocytes

    Appari Mahesh; Cario Gunnar; Richter-Unruh Annette; Demeter Janos; Werner Ralf; Bebermeier Jan-Hendrik; Holterhus Paul-Martin; Siebert Reiner; Riepe Felix; Brooks James D; Hiort Olaf


    Abstract Background Gender appears to be determined by independent programs controlled by the sex-chromosomes and by androgen-dependent programming during embryonic development. To enable experimental dissection of these components in the human, we performed genome-wide profiling of the transcriptomes of peripheral blood mononuclear cells (PBMC) in patients with rare defined "disorders of sex development" (DSD, e.g., 46, XY-females due to defective androgen biosynthesis) compared to normal 46...

  6. Phenotyping of leukocytes and granulocyte and monocyte phagocytic activity in the peripheral blood and uterus of cows with endometritis.

    Brodzki, P; Kostro, K; Brodzki, A; Lisiecka, U; Kurek, L; Marczuk, J


    This study was a comparative evaluation of selected immunological parameters in peripheral blood and uterine wash samples from cows with a normal postpartum period compared with cows with endometritis. We aimed to determine the usefulness of these parameters in monitoring the puerperium. In total, 40 cows were included in the study: 20 had endometritis (experimental group), and 20 did not have uterine inflammation (control group). Animals were chosen on the basis of cytological and bacteriological test results. The tests were conducted 5, 22, and 40 days postpartum. In both groups, flow cytometric analysis of the surface molecules CD4, CD8, CD21, CD25, and CD14 in the peripheral blood and uterine washings was performed. Granulocyte and monocyte phagocytic activity was determined using a commercial Phagotest kit that was adapted for flow cytometry. The percentage of phagocytic granulocytes and monocytes in both the peripheral blood and the uterine washings was significantly lower for cows in the experimental group compared with the control group (P cows with endometritis. A significant decrease (P endometritis. Knowledge of the immunological mechanisms observed in cows with endometritis might aid in choosing the correct immunomodulating agent-based adjuvant therapy. PMID:24857644

  7. Combined Effects of Gamma Radiation and High Dietary Iron on Peripheral Leukocyte Distribution and Function

    Crucian, Brian E.; Morgan, Jennifer L. L.; Quiriarte, Heather A.; Sams, Clarence F.; Smith, Scott M.; Zwart, Sara R.


    Both radiation and increased iron stores can independently increase oxidative damage, resulting in protein, lipid and DNA oxidation. Oxidative stress increases the risk of many health problems including cancer, cataracts, and heart disease. This study, a subset of a larger interdisciplinary investigation of the combined effect of iron overload on sensitivity to radiation injury, monitored immune parameters in the peripheral blood of rats subjected to gamma radiation, high dietary iron or both. Specific immune measures consisted of: (1) peripheral leukocyte distribution, (2) plasma cytokine levels and (3) cytokine production profiles following whole blood mitogenic stimulation

  8. Identification of an additional class of C3-binding membrane proteins of human peripheral blood leukocytes and cell lines.

    Cole, J L; Housley, G A; Dykman, T R; MacDermott, R P; Atkinson, J P


    Proteins binding the third component of complement (C3) were isolated by affinity chromatography from surface-labeled solubilized membranes of human peripheral blood cells and cell lines. The isolated molecules were subjected to NaDodSO4/PAGE, and autoradiographs of these gels indicated that C3-binding proteins could be divided into three groups based on Mr: (i) gp200, an approximately 200,000 Mr molecule previously identified as the C3b/C4b receptor or CR1; (ii) gp140, an approximately 140,0...

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

    Rui-Ju Niu


    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.

  10. West Nile virus-induced cell adhesion molecules on human brain microvascular endothelial cells regulate leukocyte adhesion and modulate permeability of the in vitro blood-brain barrier model.

    Kelsey Roe

    Full Text Available Characterizing the mechanisms by which West Nile virus (WNV causes blood-brain barrier (BBB disruption, leukocyte infiltration into the brain and neuroinflammation is important to understand the pathogenesis of WNV encephalitis. Here, we examined the role of endothelial cell adhesion molecules (CAMs in mediating the adhesion and transendothelial migration of leukocytes across human brain microvascular endothelial cells (HBMVE. Infection with WNV (NY99 strain significantly induced ICAM-1, VCAM-1, and E-selectin in human endothelial cells and infected mice brain, although the levels of their ligands on leukocytes (VLA-4, LFA-1and MAC-1 did not alter. The permeability of the in vitro BBB model increased dramatically following the transmigration of monocytes and lymphocytes across the models infected with WNV, which was reversed in the presence of a cocktail of blocking antibodies against ICAM-1, VCAM-1, and E-selectin. Further, WNV infection of HBMVE significantly increased leukocyte adhesion to the HBMVE monolayer and transmigration across the infected BBB model. The blockade of these CAMs reduced the adhesion and transmigration of leukocytes across the infected BBB model. Further, comparison of infection with highly neuroinvasive NY99 and non-lethal (Eg101 strain of WNV demonstrated similar level of virus replication and fold-increase of CAMs in HBMVE cells suggesting that the non-neuropathogenic response of Eg101 is not because of its inability to infect HBMVE cells. Collectively, these results suggest that increased expression of specific CAMs is a pathological event associated with WNV infection and may contribute to leukocyte infiltration and BBB disruption in vivo. Our data further implicate that strategies to block CAMs to reduce BBB disruption may limit neuroinflammation and virus-CNS entry via 'Trojan horse' route, and improve WNV disease outcome.

  11. The expression of GPR109A, NF-kB and IL-1β in peripheral blood leukocytes from patients with type 2 diabetes.

    Liu, Fengxiu; Fu, Yucai; Wei, Chiju; Chen, Yongru; Ma, Shuhua; Xu, Wencan


    This study was designed to explore the association between the G protein-coupled receptor 109A (GPR109A) expression in peripheral blood leukocytes (PBLs) and type 2 diabetes (T2DM) and to discuss the regulation of inflammatory factors by GPR109A signaling. GPR109A signaling has been confirmed to be associated with homeostasis of glucose/lipid metabolism, but the role of signaling in T2DM is still poorly understood. Peripheral blood samples and biochemical data were collected from healthy individuals (normal controls) and T2DM patients. Immunocytochemical staining was used to detect the expression of GPR109A in PBLs. Reverse transcription polymerase chain reaction (RT-PCR) was used to measure mRNA levels of GPR109A, NF-κB, and IL-1β in PBLs. Immunocytochemical staining showed that the GPR109A protein is localized in the nucleus and cytoplasm of granulocytes, monocytes, and lymphocytes. RT-PCR showed that mRNA levels of GPR109A, NF-κB, and IL-1β were higher in the T2DM group than in the control group (P<0.05). Correlation analysis showed a positive correlation both between GPR109A/NF-κB (r=0.376, P<0.05), and GPR109A/IL-1β (r=501, P<0.05) and between GPR109A and fasting plasma glucose (FPG) (r=0.179, P<0.05) and NF-κB /FPG (r=0.358, p<0.05). Our results suggest that GPR109A signaling is associated with T2DM, playing a role in regulation of the inflammatory cytokines. PMID:25361930

  12. Towards hemerythrin-based blood substitutes: Comparative performance to hemoglobin on human leukocytes and umbilical vein endothelial cells

    Eva Fischer-Fodor; Augustin Mot; Florina Deac; Mariann Arkosi; Radu Silaghi-Dumitrescu


    Hemerythrin is a dioxygen-carrying protein whose oxidative/nitrosative stress-related reactivity is lower than that of hemoglobin, which may warrant investigation of hemerythrin as raw material for artificial oxygen carriers (‘blood substitutes’). We report here the first biological tests for hemerythrin and its chemical derivatives, comparing their performance with that of a representative competitor, glutaraldehyde-polymerized bovine hemoglobin. Hemerythrin (native or derivatized) exhibits a proliferative effect on human umbilical vein endothelial cell (HUVEC) cultures, as opposed to a slight inhibitory effect of hemoglobin. A similar positive effect is displayed on human lymphocytes by glutaraldehyde-polymerized hemerythrin, but not by native or polyethylene glycol-derivatized hemerythrin.

  13. Preparation of 111In leukocytes after hemolytic removal of erythrocytes

    An optimized procedure is described for isolation and high-efficiency radiolabeling of leukocytes using 111In-oxine. The chief advantages over conventional methods include virtually no loss of leukocytes during washing and separation steps; a significant reduction in the time required to prepare leukocytes for radiolabeling compared to non-hemolytic preparations; a 28% increase in the average labeling efficiency obtained using 111In-oxine; > 95% cell viability as measured by the trypan blue exclusion test; elimination of contaminating red blood cells from the leukocyte pellet prior to labeling; and 80% survivability at 15 min post injection (measured as per cent of blood activity on leukocyte fraction). (author)

  14. Diagnostic value of serum procalcitonin levels in children with meningitis: a comparison with blood leukocyte count and C-reactive protein

    Objectives: To determine the level of serum procalcitonin, blood leukocyte count (TLC) and C-reactive protein (CRP) in children with bacterial and non bacterial meningitis and document their efficacy in differential diagnosis. Also described are procalcitonin levels variation during treatment. Methods: From March 2005 to February 2008, we evaluated 38 clinically suspected meningitis patients in the paediatric departments, Al-Jedaany Hospital, Jeddah, KSA, for Serum procalcitonin, CRP, TLC and Lumbar punctures and CSF analysis. Patients were classified into bacterial meningitis group I (18) and non bacterial meningitis group II (20). Results: Serum PCT levels were significantly higher in bacterial meningitis (BM) 9 mean 4.8 +- 3.85 ng/ml (2.9-11.6)) compared with non bacterial meningitis (NBM) (mean 0.38 +- 0.25 ng/ml(0.31-0.61)) P< 0.001). Mean of all CSF parameters, TLC (15,000 +- 2,900 cell/ml(BM) and 9,500 +-1,105 cell/ml(NBM))and CRP (20 +- 6.8 mg/l (BM) and 12.5 +-12.0 mg/l(NBM))showed a zone of overlapping between the two groups. There is a positive correlation between serum PCT, TLC and CRP in bacterial and non bacterial meningitis cases but this relation becomes highly significant with bacterial meningitis positive group. Day 3 and day 6 treatment serum PCT was less than on admission levels (P<0.001). Conclusion: PCT can be used in the early diagnosis of bacterial meningitis and may be a useful adjunct in differentiating bacterial and non bacterial meningitis than CRP or TLC and diminishing the value of lumbar puncture performed 48-72 hours after admission to assess treatment efficacy. (author)

  15. Separation of uncompromised whole blood mixtures for single source STR profiling using fluorescently-labeled human leukocyte antigen (HLA) probes and fluorescence activated cell sorting (FACS).

    Dean, Lee; Kwon, Ye Jin; Philpott, M Katherine; Stanciu, Cristina E; Seashols-Williams, Sarah J; Dawson Cruz, Tracey; Sturgill, Jamie; Ehrhardt, Christopher J


    Analysis of biological mixtures is a significant problem for forensic laboratories, particularly when the mixture contains only one cell type. Contributions from multiple individuals to biologic evidence can complicate DNA profile interpretation and often lead to a reduction in the probative value of DNA evidence or worse, its total loss. To address this, we have utilized an analytical technique that exploits the intrinsic immunological variation among individuals to physically separate cells from different sources in a mixture prior to DNA profiling. Specifically, we applied a fluorescently labeled antibody probe to selectively bind to one contributor in a mixture through allele-specific interactions with human leukocyte antigen (HLA) proteins that are expressed on the surfaces of most nucleated cells. Once the contributor's cells were bound to the probe, they were isolated from the mixture using fluorescence activated cell sorting (FACS)-a high throughput technique for separating cell populations based on their optical properties-and then subjected to STR analysis. We tested this approach on two-person and four-person whole blood mixtures where one contributor possessed an HLA allele (A*02) that was not shared by other contributors to the mixture. Results showed that hybridization of the mixture with a fluorescently-labeled antibody probe complimentary to the A*02 allele's protein product created a cell population with a distinct optical profile that could be easily differentiated from other cells in the mixture. After sorting the cells with FACS, genetic analysis showed that the STR profile of this cell population was consistent with that of the contributor who possessed the A*02 allele. Minor peaks from the A*02 negative contributor(s) were observed but could be easily distinguished from the profile generated from A*02 positive cells. Overall, this indicates that HLA antibody probes coupled to FACS may be an effective approach for generating STR profiles of

  16. A scintiscan study using leukocytes

    A method of labelling leukocytes with 111-In-oxine is described that is easy to handle, reproducible and requires no sophisticated hardware. It is based on a leukocyte concentrate prepared from whole blood, which has an average cell content of 64.8%. Seventy-two patients selected from among those undergoing nephrologico-urological and neurological procedures or accident surgery were subjected to preliminary wholebody scintiscanning as well as a scintiscan study using leukocytes. It was calculated that the method is able to reveal the presence of an abscess with a sensitivity of 77%, a specificity of 98% and an accuracy of 89%. This examination may prove equally useful to follow up a confirmed inflammation, to determine its exact type and extent and to make a clear diagnostic distinction between an inflammatory process and a tumour. (TRV)

  17. Zinc supplementation of young men alters metallothionein, zinc transporter, and cytokine gene expression in leukocyte populations

    Aydemir, Tolunay Beker; Blanchard, Raymond K.; Cousins, Robert J.


    An effective measure to assess zinc status of humans has remained elusive, in contrast to iron, where a number of indicators of metabolism/function are available. Using monocytes, T lymphocytes, and granulocytes isolated by magnetic sorting and dried blood spots (DBS) derived from 50 μl of peripheral blood, we evaluated the response of metallothionein (MT), zinc transporter, and cytokine genes to a modest (15 mg of Zn per day) dietary zinc supplement in human subjects. Transcript abundance was measured by quantitative real-time RT-PCR (QRT-PCR). Zinc supplementation increased MT mRNA abundance by up to 2-fold in RNA from leukocyte subsets, and 4-fold in RNA from DBS. Transcript levels for the zinc transporter genes ZnT1 and Zip3 were increased and decreased, respectively, by zinc supplementation. Expression of the ZnT and Zip genes among leukocyte subsets differ by up to 270-fold. Monocytes and granulocytes from supplemented subjects were activated by LPS, whereas T lymphocytes were activated by mimicking antigen presentation. With zinc consumption, TNF-α and IL-1β expression was greater in activated monocytes and granulocytes, and IFN-γ mRNA levels were higher in activated T lymphocytes. These studies show that QRT-PCR is a tool to reliably measure transcript abundance for nutritionally responsive genes in human subjects, and that a small sample of whole dried blood, when appropriately collected, can be used as the source of total RNA for QRT-PCR analysis. The results obtained also show that zinc supplementation of human subjects programs specific leukocytic subsets to show enhanced cytokine expression upon activation by stimulators of immunity. PMID:16434472

  18. Comparing Electrolytes in Prestorage Leukocyte-Reduced Packed Cell versus Unfiltered Packed Cell

    Fallahi, L; Ghiliyan, R; Hashemi, A; A. Fatemi; M. Saeedi


    Background Blood transfusion is associated with side effects caused by residual leukocytes in blood and blood components. Using leukodepleted blood components can decrease some of these adverse effects. Among the various methods to remove leukocytes in blood components, prestorage leukoreduction has been most efficient, but the evidence of clinical side effects awaits further studies. We evaluated changes of electrolytes in prestorage leukocyte-reduced red blood cells. Materials and Methods I...

  19. Substitution of Aspartate for glycine 1018 in the Type III procollagen (COL3AI) gene causes type IV Ehlers-Danlos Syndrome: The mutated allele is present in most blood leukocytes of the asymptomatic and mosaic mother

    Kontusaari, S.; Tromp, G.; Kuivaniemi, H.; Prockop, D.J. (Thomas Jefferson Univ., Philadelphia, PA (United States)); Stolle, C. (Robert Wood Johnson Medical School, Piscataway, NJ (United States)); Pope, F.M.


    A proband with arterial ruptures and skin changes characteristic of the type IV variant of Ehlers-Danlos syndrome was found to have a single-base mutation in the type III procollagen gene, which converted the codon for glycine at amino position 1018 to a codon for aspartate. (Amino acid positions are numbered by the standard convention in which the first glycine of the triple-helical domain of an [alpha] chain is number 1. The numbers of positions in the [alpha]1(III) chains can be converted to positions in the human pro[alpha](III) chain by adding 167.). Nucleotide sequencing of overlapping PCR products in which the two alleles were distinguished demonstrated that the mutation of glycine 1018 was the only mutation that changed the primary structure of type III procollagen. The glycine substitution markedly decreased the amount of type III procollagen secreted into the medium by cultured skin fibroblasts from the proband. It is surprising that the same mutation was found in about 94% of the peripheral blood leukocytes from the proband's asymptomatic 72-year-old mother. Other tissues from the mother contained the mutated allele; it was present in 0%-100% of different samples of hair cells and in about 40% of cells from the oral epithelium. Therefore, the mother was a mosaic for the mutation. Since the mutated allele was present in cells derived from all three germ layers, the results indicated that the mutation arose by the late blastocyst stage of development. The results also indicate that assays of blood leukocytes do not always reveal mosaicism or predict phenotypic involvement of tissues, such as blood vessels, that are derived from the same embryonic cells as are leukocytes. 66 refs., 6 figs., 1 tab.

  20. Interactions Between Stably Rolling Leukocytes In Vivo

    King, M R; Kim, M B; Sarelius, I H; King, Michael R.; Ruscio, Aimee D.; Kim, Michael B.; Sarelius, Ingrid H.


    We have characterized the two-dimensional spatial dependence of the hydrodynamic interactions between two adhesively rolling leukocytes in a live venule in the mouse cremaster muscle. Two rolling leukocytes were observed to slow each other down when rolling together in close proximity, due to mutual sheltering from the external blood flow in the vessel lumen. These results are in agreement with a previous study of leukocyte rolling interactions using carbohydrate-coated beads in a parallel-plate flow chamber and a detailed computer model of adhesion in a multicellular environment.

  1. Effects of ochratoxin a on broiler leukocytes

    MA Moura


    Full Text Available This study evaluated alterations in the qualitative cellular profile of leukocytes caused by the administration of low doses of ochratoxin-A (OTA in poultry. Sixty chicks were separated in three experimental groups: control, PBS-treated and OTA-treated. Blood smears from all birds were analyzed three and six hours post-treatment. Differential leukocyte counting demonstrated that OTA reduced the percentage of lymphocytes and eosinophils and significantly increased the number of heterophils and monocytes.

  2. Leukocyte nucleus segmentation and nucleus lobe counting

    Zheng Zong-Han


    Full Text Available Abstract Background Leukocytes play an important role in the human immune system. The family of leukocytes is comprised of lymphocytes, monocytes, eosinophils, basophils, and neutrophils. Any infection or acute stress may increase or decrease the number of leukocytes. An increased percentage of neutrophils may be caused by an acute infection, while an increased percentage of lymphocytes can be caused by a chronic bacterial infection. It is important to realize an abnormal variation in the leukocytes. The five types of leukocytes can be distinguished by their cytoplasmic granules, staining properties of the granules, size of cell, the proportion of the nuclear to the cytoplasmic material, and the type of nucleolar lobes. The number of lobes increased when leukemia, chronic nephritis, liver disease, cancer, sepsis, and vitamin B12 or folate deficiency occurred. Clinical neutrophil hypersegmentation has been widely used as an indicator of B12 or folate deficiency.Biomedical technologists can currently recognize abnormal leukocytes using human eyes. However, the quality and efficiency of diagnosis may be compromised due to the limitations of the biomedical technologists' eyesight, strength, and medical knowledge. Therefore, the development of an automatic leukocyte recognition system is feasible and necessary. It is essential to extract the leukocyte region from a blood smear image in order to develop an automatic leukocyte recognition system. The number of lobes increased when leukemia, chronic nephritis, liver disease, cancer, sepsis, and vitamin B12 or folate deficiency occurred. Clinical neutrophil hypersegmentation has been widely used as an indicator of B12 or folate deficiency. Results The purpose of this paper is to contribute an automatic leukocyte nuclei image segmentation method for such recognition technology. The other goal of this paper is to develop the method of counting the number of lobes in a cell nucleus. The experimental results


    南克俊; 李春丽; 魏永长; 隋晨光; 刘亚民; 陈葳


    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.

  4. Maternal BMI Associations with Maternal and Cord Blood Vitamin D Levels in a North American Subset of Hyperglycemia and Adverse Pregnancy Outcome (HAPO Study Participants.

    Jami L Josefson

    Full Text Available Obesity in pregnancy may be associated with reduced placental transfer of 25-hydroxyvitamin D (25-OHD. The objective of this study was to examine associations between maternal BMI and maternal and cord blood levels of 25-OHD in full term neonates born to a single racial cohort residing at similar latitude. Secondary objectives were to examine associations between maternal glucose tolerance with maternal levels of 25-OHD and the relationship between cord blood 25-OHD levels and neonatal size.This study was conducted among participants of the Hyperglycemia and Adverse Pregnancy Outcomes (HAPO Study meeting the following criteria: residing at latitudes 41-43°, maternal white race, and gestational age 39-41 weeks. Healthy pregnant women underwent measures of height, weight, and a 75-g fasting oral glucose tolerance test (OGTT at approximately 28 weeks gestation. Maternal and cord blood sera were analyzed for total 25-OHD by HPLC tandem mass spectrometry. Statistical analyses included ANOVA and linear regression models.Maternal and cord blood (N = 360 mean levels (sd of 25-OHD were 37.2 (11.2 and 23.4 (9.2 ng/ml, respectively, and these levels were significantly different among the 3 field centers (ANOVA p< 0.001. Maternal serum 25-OHD was lower by 0.40 ng/ml for BMI higher by 1 kg/m2 (p<0.001 in an adjusted model. Maternal fasting plasma glucose, insulin sensitivity, and presence of GDM were not associated with maternal serum 25-OHD level when adjusted for maternal BMI. Cord blood 25-OHD was lower by 0.26 ng/ml for maternal BMI higher by 1 kg/m2 (p<0.004. With adjustment for maternal age, field center, birth season and maternal serum 25-OHD, the association of cord blood 25-OHD with maternal BMI was attenuated. Neither birth weight nor neonatal adiposity was significantly associated with cord blood 25-OHD levels.These results suggest that maternal levels of 25-OHD are associated with maternal BMI. The results also suggest that interpretation of

  5. Remodeling of B-Cell Subsets in Blood during Pegylated IFNα-2a Therapy in Patients with Chronic Hepatitis B Infection

    Jacob, Marie-Christine; Dufeu-Duchesne, Tania; Bertucci, Inga; Pouget, Noelle; Brevot-Lutton, Ophelie; Zoulim, Fabien; Bourliere, Marc; Plumas, Joel; Leroy, Vincent


    The ultimate goal of pegylated interferon-alfa-2a (Peg-IFN-α) therapy in chronic hepatitis B (CHB) infection is HBsAg seroconversion. Even though B cells are major mediators of a positive clinical outcome, their modulation during Peg-IFN-α therapy has not yet been described. We investigated here the effects of Peg-IFN-α on eight circulating B-cell subsets thanks to an original multi-gating approach based on CD19, CD27, IgD, CD10, and CD38 markers in patients with CHB treated with nucleos(t)ide analog alone or in combination with Peg-IFN-α. These dynamic changes were analyzed during the 48-weeks of Peg-IFN-α therapy and up to 2 years after the cessation of treatment. The CD19+CD27-IgD+CD10+CD38high transitional B cells and the CD19+CD27+IgD-CD10-CD38high plasmablasts continuously increased, whereas the CD19+CD27-IgD+CD10-CD38low naive, CD19+CD27+IgD+ natural memory, and CD19+CD27+IgD-CD10-CD38low post-germinal center B cells decreased during the course of Peg-IFNα treatment. Such modulations correlated with a sustained increase in sCD30 levels and the decrease in plasma HBsAg. However, no seroconversion occurred and all parameters returned to baseline after the stop of the treatment. Peg-IFN-α therapy mediates a remodeling of B-cell compartmentalization, without clinical relevance. Our study provides new insights into the immunomodulatory effects of Peg-IFN-α on circulating B-cells, and questioned the benefit of the add-on Peg-IFN-α treatment in CHB. PMID:27281019

  6. Remodeling of B-Cell Subsets in Blood during Pegylated IFNα-2a Therapy in Patients with Chronic Hepatitis B Infection.

    Aspord, Caroline; Bruder Costa, Juliana; Jacob, Marie-Christine; Dufeu-Duchesne, Tania; Bertucci, Inga; Pouget, Noelle; Brevot-Lutton, Ophelie; Zoulim, Fabien; Bourliere, Marc; Plumas, Joel; Leroy, Vincent


    The ultimate goal of pegylated interferon-alfa-2a (Peg-IFN-α) therapy in chronic hepatitis B (CHB) infection is HBsAg seroconversion. Even though B cells are major mediators of a positive clinical outcome, their modulation during Peg-IFN-α therapy has not yet been described. We investigated here the effects of Peg-IFN-α on eight circulating B-cell subsets thanks to an original multi-gating approach based on CD19, CD27, IgD, CD10, and CD38 markers in patients with CHB treated with nucleos(t)ide analog alone or in combination with Peg-IFN-α. These dynamic changes were analyzed during the 48-weeks of Peg-IFN-α therapy and up to 2 years after the cessation of treatment. The CD19+CD27-IgD+CD10+CD38high transitional B cells and the CD19+CD27+IgD-CD10-CD38high plasmablasts continuously increased, whereas the CD19+CD27-IgD+CD10-CD38low naive, CD19+CD27+IgD+ natural memory, and CD19+CD27+IgD-CD10-CD38low post-germinal center B cells decreased during the course of Peg-IFNα treatment. Such modulations correlated with a sustained increase in sCD30 levels and the decrease in plasma HBsAg. However, no seroconversion occurred and all parameters returned to baseline after the stop of the treatment. Peg-IFN-α therapy mediates a remodeling of B-cell compartmentalization, without clinical relevance. Our study provides new insights into the immunomodulatory effects of Peg-IFN-α on circulating B-cells, and questioned the benefit of the add-on Peg-IFN-α treatment in CHB. PMID:27281019

  7. Interleukin 2 (IL 2) up-regulates its own receptor on a subset of human unprimed peripheral blood lymphocytes and triggers their proliferation

    Several reports indicate that human peripheral blood lymphoctyes (PBL) seeded in culture with purified or recombinant interleukin 2 (IL 2) immediately after separation from the blood display a substantial level of proliferation at day 5 or 6, even in the absence of any activating signal. The spontaneously IL 2 proliferating cells are large lymphocytes, and they co-purify on a Percoll gradient in the large granular lymphocytes (third (LGL) fraction) together with the natural killer (NK) activity. When LGL were separated into NKH1 (an NK-specific surface marker)-positive and NKH1-negative cells by fluorescence-activated cell sorting (FACS), proliferating cells were mainly found in the NKH1-negative fraction. On the contrary, when cells from Percoll fraction 3 were separated into OKT3-negative and positive cells, the majority of the proliferating cells was found in the OKT3-positive cells. These results indicate that spontaneously IL 2 proliferating (SIP) cells most probably belong to the T cell lineage, but are distinct from NK cells. Additional analysis of Il 2 receptor induced in culture with IL 2 was performed by [125I]anti-TAC binding and by [3H]Il 2 binding. Scatchard analysis of [3H]IL 2 binding, in the range of concentrations leading to the detection of high-affinity binding sites, showed an affinity constant similar to that of conventional phytohemagglutinin blasts. The results indicate that SIP cells are preactivated cells circulating in the blood. They are large cells and represent a very small proportion of circulating lymphocytes (0.3%). They express a subliminar amount of IL 2 receptor. Cultivated in the presence of IL 2, IL 2 receptor expression is enhanced to a detectable level, and the SIP cells begin to proliferate. These SIP cells could be activated T cells in the course of a current immune response or memory T cells present in every normal individual

  8. Factoring groups into subsets

    Szabo, Sandor


    Decomposing an abelian group into a direct sum of its subsets leads to results that can be applied to a variety of areas, such as number theory, geometry of tilings, coding theory, cryptography, graph theory, and Fourier analysis. Focusing mainly on cyclic groups, Factoring Groups into Subsets explores the factorization theory of abelian groups. The book first shows how to construct new factorizations from old ones. The authors then discuss nonperiodic and periodic factorizations, quasiperiodicity, and the factoring of periodic subsets. They also examine how tiling plays an important role in n

  9. Real-time imaging reveals the dynamics of leukocyte behaviour during experimental cerebral malaria pathogenesis.

    Saparna Pai


    Full Text Available During experimental cerebral malaria (ECM mice develop a lethal neuropathological syndrome associated with microcirculatory dysfunction and intravascular leukocyte sequestration. The precise spatio-temporal context in which the intravascular immune response unfolds is incompletely understood. We developed a 2-photon intravital microscopy (2P-IVM-based brain-imaging model to monitor the real-time behaviour of leukocytes directly within the brain vasculature during ECM. Ly6C(hi monocytes, but not neutrophils, started to accumulate in the blood vessels of Plasmodium berghei ANKA (PbA-infected MacGreen mice, in which myeloid cells express GFP, one to two days prior to the onset of the neurological signs (NS. A decrease in the rolling speed of monocytes, a measure of endothelial cell activation, was associated with progressive worsening of clinical symptoms. Adoptive transfer experiments with defined immune cell subsets in recombinase activating gene (RAG-1-deficient mice showed that these changes were mediated by Plasmodium-specific CD8(+ T lymphocytes. A critical number of CD8(+ T effectors was required to induce disease and monocyte adherence to the vasculature. Depletion of monocytes at the onset of disease symptoms resulted in decreased lymphocyte accumulation, suggesting reciprocal effects of monocytes and T cells on their recruitment within the brain. Together, our studies define the real-time kinetics of leukocyte behaviour in the central nervous system during ECM, and reveal a significant role for Plasmodium-specific CD8(+ T lymphocytes in regulating vascular pathology in this disease.

  10. File list: InP.Bld.50.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Full Text Available InP.Bld.50.AllAg.Polymorphonuclear_leukocytes hg19 Input control Blood Polymorphonuclear... leukocytes ...

  11. File list: ALL.Bld.50.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

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  12. File list: ALL.Bld.05.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

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  13. File list: NoD.Bld.20.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

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  14. File list: InP.Bld.20.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Full Text Available InP.Bld.20.AllAg.Polymorphonuclear_leukocytes hg19 Input control Blood Polymorphonuclear... leukocytes ...

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  18. File list: ALL.Bld.10.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

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  1. File list: ALL.Bld.20.AllAg.Polymorphonuclear_leukocytes [Chip-atlas[Archive

    Full Text Available ALL.Bld.20.AllAg.Polymorphonuclear_leukocytes hg19 All antigens Blood Polymorphonuclear... leukocytes SRX1016682,SRX1016679,SRX1016680,SRX1016681 ...

  2. Microarray analysis of human leucocyte subsets: the advantages of positive selection and rapid purification

    Chaudhry Afzal N


    Full Text Available Abstract Background For expression profiling to have a practical impact in the management of immune-related disease it is essential that it can be applied to peripheral blood cells. Early studies have used total peripheral blood mononuclear cells, and as a consequence the majority of the disease-related signatures identified have simply reflected differences in the relative abundance of individual cell types between patients and controls. To identify cell-specific changes in transcription it would be necessary to profile purified leucocyte subsets. Results We have used sequential rounds of positive selection to isolate CD4 and CD8 T cells, CD19 B cells, CD14 monocytes and CD16 neutrophils for microarray analysis from a single blood sample. We compared gene expression in cells isolated in parallel using either positive or negative selection and demonstrate that there are no significant consistent changes due to positive selection, and that the far inferior results obtained by negative selection are largely due to reduced purity. Finally, we demonstrate that storing cells prior to separation leads to profound changes in expression, predominantly in cells of the myeloid lineage. Conclusion Leukocyte subsets should be prepared for microarray analysis by rapid positive selection.

  3. Unsupervised Feature Subset Selection

    Søndberg-Madsen, Nicolaj; Thomsen, C.; Pena, Jose

    This paper studies filter and hybrid filter-wrapper feature subset selection for unsupervised learning (data clustering). We constrain the search for the best feature subset by scoring the dependence of every feature on the rest of the features, conjecturing that these scores discriminate some...... irrelevant features. We report experimental results on artificial and real data for unsupervised learning of naive Bayes models. Both the filter and hybrid approaches perform satisfactorily....

  4. On best subset regression

    Xiong, Shifeng


    In this paper we discuss the variable selection method from \\ell0-norm constrained regression, which is equivalent to the problem of finding the best subset of a fixed size. Our study focuses on two aspects, consistency and computation. We prove that the sparse estimator from such a method can retain all of the important variables asymptotically for even exponentially growing dimensionality under regularity conditions. This indicates that the best subset regression method can efficiently shri...

  5. Leukocyte Adhesion Deficiency (LAD)

    ... Content Marketing Share this: Main Content Area Leukocyte Adhesion Deficiency (LAD) LAD is an immune deficiency in ... are slow to heal also may have LAD. Treatment and Research Doctors prescribe antibiotics to prevent and ...

  6. Leukocytic acetylcholine in chronic rejection of renal allografts

    Wilczynska, Joanna


    Leukocytes, which accumulate in graft blood vessels during fatal acute rejection of experimental renal allografts, synthesise and release acetylcholine (ACh). In this study, I tested the hypothesis that ACh produced by leukocytes accumulating in graft blood vessels contributes to the pathogenesis of chronic renal allograft vasculopathy (CAV). Kidneys were transplanted in the allogeneic Fischer 344 to Lewis rat strain combination. Isogeneic transplantations were performed in Lew...

  7. Allogeneic leukocytes in cardiac surgery: Good or bad?

    Anneke Brand


    Full Text Available Worldwide, cardiac surgery is a common procedure requiring a large quantity of allogeneic blood products, which are associated with postoperative complications. Leukocytes present in blood products may play a role in these complications, which are referred to as transfusion-related immunomodulation (TRIM. Several randomized controlled trials (RCTs in different settings investigated the effects of allogeneic leukocytes in red blood cells (RBCs. Cardiac surgery studies reported a reduction in postoperative infections and mortality in patients that received leukocyte-reduced RBCs compared with leukocyte-containing RBCs; this was mainly due to more deaths due to infections and multiple organ dysfunction syndrome (MODS in the group that received leukocyte-containing RBCs. Patients with postoperative complications had higher concentrations of inflammatory mediators. These findings suggest that leukocyte-containing transfusion during cardiac surgery induces a second insult to the systemic inflammatory response. In the present review we discuss the possible role of blood transfusions in cardiac surgery. Especially, we focus on the possible role of allogeneic leukocytes associated with postoperative complications after cardiac surgery.

  8. Electrophoretic detection of protein p53 in human leukocytes

    The authors have found an acid-soluble protein with mol. wt. of about 53 kD in peripheral blood leukocytes of persons with Down's syndrome. It was present in different quantities in all 20 patients tested, but was virtually not discovered in 12 healthy blood donors. This paper determines the possible identity of this protein with protein p53 from mouse ascites carcinoma by comparing their electrophoretic mobilities, because the accuracy of electrophoretic determination of the molecular weight of proteins is not sufficient to identify them. The paper also describes experiments to detect a protein with electrophoretic mobility identical with that of a protein in the leukocytes of patients with Down's syndrome in leukocytes of patients with leukemia. To discover if protein p53 is involved in cell proliferation, the protein composition of leukocytes from healthy blood donors, cultured in the presence and absence of phytohemagglutinin (PHA), was compared. Increased incorporation of H 3-thymidine by leukocytes of patients with Down's syndrome is explained by the presence of a population of immature leukocytes actively synthesizing DNA in the peripheral blood of these patients, and this can also explain the presence of protein p53 in the leukocytes of these patients

  9. A Progenitor Cell Expressing Transcription Factor RORγt Generates All Human Innate Lymphoid Cell Subsets.

    Scoville, Steven D; Mundy-Bosse, Bethany L; Zhang, Michael H; Chen, Li; Zhang, Xiaoli; Keller, Karen A; Hughes, Tiffany; Chen, Luxi; Cheng, Stephanie; Bergin, Stephen M; Mao, Hsiaoyin C; McClory, Susan; Yu, Jianhua; Carson, William E; Caligiuri, Michael A; Freud, Aharon G


    The current model of murine innate lymphoid cell (ILC) development holds that mouse ILCs are derived downstream of the common lymphoid progenitor through lineage-restricted progenitors. However, corresponding lineage-restricted progenitors in humans have yet to be discovered. Here we identified a progenitor population in human secondary lymphoid tissues (SLTs) that expressed the transcription factor RORγt and was unique in its ability to generate all known ILC subsets, including natural killer (NK) cells, but not other leukocyte populations. In contrast to murine fate-mapping data, which indicate that only ILC3s express Rorγt, these human progenitor cells as well as human peripheral blood NK cells and all mature ILC populations expressed RORγt. Thus, all human ILCs can be generated through an RORγt(+) developmental pathway from a common progenitor in SLTs. These findings help establish the developmental signals and pathways involved in human ILC development. PMID:27178467

  10. Identification and characterization of human dendritic cell subsets in the steady state: a review of our current knowledge.

    Patel, Vineet Indrajit; Metcalf, Jordan Patrick


    Dendritic cells (DC) are generally categorized as a group of rare antigen presenting cells that are to the crucial development of immune responses to pathogens and also of tolerance to self-antigens. Therefore, having the ability to identify DC in specific tissues and to test their functional abilities in the steady state are scientific gaps needing attention. Research on primary human DC is lacking due to their rarity and the difficulty of obtaining tissue samples. However, recent findings have shown that several different DC subsets exist, and that these subsets vary both by markers expressed and functions depending on their specific microenvironment. After discriminating from other cell types, DC can be split into myeloid and plasmacytoid fractions. While plasmacytoid DC express definite markers, CD123 and BDCA-2, myeloid DC encompass several different subsets with overlapping markers expressed. Such markers include the blood DC antigens BDCA-1 and BDCA-3, along with Langerin, CD1a and CD14. Marker specificity is further reduced when accounting for microenvironmental differences, as observed in the blood, primary lymphoid tissues, skin and lungs. The mixed leukocyte reaction (MLR) has been used to measure the strength of antigen presentation by specific DC subsets. Surface markers and MLR require standardization to enable consistent identification of and comparisons between DC subsets. To alleviate these issues, researchers have begun comparing DC subsets at the transcriptional level. This has allowed degrees of relatedness to be determined between DC in different microenvironments, and should be a continued area of focus in years to come. PMID:26956785

  11. Subset selection in regression

    Miller, Alan


    Originally published in 1990, the first edition of Subset Selection in Regression filled a significant gap in the literature, and its critical and popular success has continued for more than a decade. Thoroughly revised to reflect progress in theory, methods, and computing power, the second edition promises to continue that tradition. The author has thoroughly updated each chapter, incorporated new material on recent developments, and included more examples and references. New in the Second Edition:A separate chapter on Bayesian methodsComplete revision of the chapter on estimationA major example from the field of near infrared spectroscopyMore emphasis on cross-validationGreater focus on bootstrappingStochastic algorithms for finding good subsets from large numbers of predictors when an exhaustive search is not feasible Software available on the Internet for implementing many of the algorithms presentedMore examplesSubset Selection in Regression, Second Edition remains dedicated to the techniques for fitting...

  12. Clinical evaluation of leukocyte differential count in peripheral blood by five-color flow cytometry%五色流式细胞术分类计数外周血白细胞的临床评价

    邢莹; 王建中; 普程伟; 尚柯


    目的 评价五色流式细胞术分类计数外周血白细胞的临床应用价值.方法 选取2010年9至12月北京大学第一医院就诊的265例患者静脉全血标本,以显微镜检查为参考方法,同时用LH750全自动血细胞分析仪和FC500流式细胞仪分类计数白细胞,统计分析显微镜、血细胞分析仪和流式细胞术分类计数外周血白细胞的相关性.选取2010年8至11月北京大学第一医院、北京市道培医院和北京军区总医院的40例患者的静脉全血标本,以显微镜检查为参考方法,同时用LH750全自动血细胞分析仪和NAVIOS流式细胞仪分类计数白细胞,统计分析显微镜、血细胞分析仪和流式细胞术分类计数外周血白细胞的相关性和流式细胞术的临床诊断效率.结果 FC500流式细胞仪采用仪器自动分析软件进行分类,与显微镜分类计数5种白细胞均具有一定相关性(均P<0.01),其中以淋巴细胞、中性粒细胞和嗜酸粒细胞的相关性最佳(r=0.955、0.928、0.876).NAVIOS 流式细胞仪采用手工输入设门策略进行分类,与显微镜方法比较除嗜碱粒细胞外均具有良好的相关性(r >0.700,均P<0.01),其中对检出中性粒细胞、淋巴细胞和原始细胞的相关性较高(r=0.950、0.915、0.852).以显微镜法分类计数外周血不成熟粒细胞1%为临界值,流式细胞术检出不成熟粒细胞的敏感度为87%,特异度为76%.以显微镜法检查外周血原始细胞0.5%为临界值,流式细胞术检出的敏感度为100%,特异度为92%.结论 应用五色流式细胞术在不同流式细胞仪上分类计数外周血白细胞与显微镜法均具有良好的相关性,对检出原始细胞和不成熟粒细胞具有较高的敏感度.%Objective To explore the clinical application values of five-color flow cytometry for leukocyte differential count in peripheral blood.Methods Leukocyte differentiation in 265 peripheral blood samples collected at

  13. In utero infection with porcine reproductive and respiratory syndrome virus modulates leukocyte subpopulations in peripheral blood and bronchoalveolar fluid of suviving piglets

    Nielsen, J.; Bøtner, Anette; Tingstedt, J. E.;


    of CD2+, CD4+CD8+ and SLA-classII+ cells, respectively, in peripheral blood, together with the levels of CD2+ and CD3+ cells in BALF were increased in the infected piglets infected in utero compared to the uninfected controls. The kinetic analyses carried out in the present study reflect that in...

  14. Human Neonatal Peripheral Blood Leukocytes Demonstrate Pathogen-Specific Coordinate Expression of TLR2, TLR4/MD2 and MyD88 During Bacterial Infection In Vivo

    Zhang, Jin-Ping; Yang, Yi(Department of Electrophysics, National Chiao-Tung University, Hsinchu, Taiwan, ROC); Levy, Ofer; Chen, Chao


    Toll-like receptors (TLRs) play important roles in infection. We have previously reported TLR2 is up-regulated in neonatal Gram-positive (G+) bacteremia whereas TLR4 is up-regulated in neonatal Gram-negative (G−) bacteremia. For functional signaling, TLR4 requires MD-2 and both TLR2 and TLR4 signal need MyD88. However, it is unknown whether newborns can enhance expression of MD-2 and MyD88 with bacterial infection in coordination with TLR expression. We characterized neonatal peripheral blood...

  15. Immune invasion of the central nervous system parenchyma and experimental allergic encephalomyelitis, but not leukocyte extravasation from blood, are prevented in macrophage-depleted mice

    Tran, E H; Hoekstra, K; van Rooijen, N;


    /J mice was abrogated by Cl2MDP-mnL treatment. CD4+ T cell and MHC II+ B220+ B cell extravasation from blood vessels and Th1 cytokine production were not inhibited. However, invasion of the central nervous system intraparenchymal tissues by lymphocytes, F4/80+, Mac-1+, and MOMA-1+ macrophages was almost....../microglial activation, was inhibited. This intervention reveals a role for macrophages in regulating the invasion of autoreactive T cells and secondary glial recruitment that ordinarily lead to demyelinating pathology in EAE and multiple sclerosis....

  16. Influence of Hydration Status on Changes in Plasma Cortisol, Leukocytes, and Antigen-Stimulated Cytokine Production by Whole Blood Culture following Prolonged Exercise.

    Svendsen, Ida S; Killer, Sophie C; Gleeson, Michael


    Elevated antigen-stimulated anti-inflammatory cytokine production appears to be a risk factor for upper respiratory tract illness in athletes. The purpose of this study was to determine the effects of prolonged exercise and hydration on antigen-stimulated cytokine production. Twelve healthy males cycled for 120 min at 60% [Formula: see text] on two occasions, either euhydrated or moderately hypohydrated (induced by fluid restriction for 24 h). Blood samples were collected before and after exercise and following 2 h recovery for determination of cell counts, plasma cortisol, and in vitro antigen-stimulated cytokine production by whole blood culture. Fluid restriction resulted in mean body mass loss of 1.3% and 3.9% before and after exercise, respectively. Exercise elicited a significant leukocytosis and elevated plasma cortisol, with no differences between trials. IL-6 production was significantly reduced 2 h postexercise (P hydration status was observed for the measured variables. Prolonged exercise appears to result in augmented anti-inflammatory cytokine release in response to antigen challenge, possibly coupled with acute suppression of proinflammatory cytokine production, corresponding with studies using mitogen or endotoxin as stimulant. Moderate hypohydration does not appear to influence these changes. PMID:24967270

  17. Effect of Transcatheter Arterial Chemoembolization Combined with Argon-Helium Cryosurgery System on the Changes of NK Cells and T Cell Subsets in Peripheral Blood of Hepatocellular Carcinoma Patients.

    Huang, Manping; Wang, Xiaoyi; Bin, Huang


    Hepatocellular carcinoma (HCC) is one of the most aggressive tumors in humans. T lymphocytes and natural killer (NK) cells are the body's first line of defense to prevent tumor cell growth. Previous studies have demonstrated that transcatheter arterial chemoembolization (TACE) combined with argon-helium cryosurgery system (AHCS) can effectively treat liver cancer. However, the mechanism of the treatment is unclear yet. In the current study, we investigated the effects of TACE combined with AHCS on the changes of T cell subsets and NK cells in peripheral blood of HCC. Our data show that alpha-fetoprotein (AFP) levels in peripheral blood were significantly up-regulated in HCC patients before treatment when compared with healthy people and reduced after TACE combined with AHCS treatment (P < 0.01). In addition, we found that CD4+ cells and NK cells decreased (P < 0.05) and CD8+ cells increased (P < 0.05) in HCC patients when compared with healthy people. After treatment, the CD4+ cells, CD4+/CD8+ ratio, and NK cells were dramatically increased in HCC patients (P < 0.05). In contrast, CD8+ cells were significantly decreased (P < 0.05). TACE combined with AHCS treatment significantly prolonged 1-year survival rate of HCC patients and did not show significant side effects. Taken together, our data indicate that TACE combined with AHCS treatment improves patients' immune system. It is a feasible and effective therapeutic method for HCC patients. PMID:27259326

  18. Rescue from acute neuroinflammation by pharmacological chemokine-mediated deviation of leukocytes

    Berghmans Nele


    Full Text Available Abstract Background Neutrophil influx is an important sign of hyperacute neuroinflammation, whereas the entry of activated lymphocytes into the brain parenchyma is a hallmark of chronic inflammatory processes, as observed in multiple sclerosis (MS and its animal models of experimental autoimmune encephalomyelitis (EAE. Clinically approved or experimental therapies for neuroinflammation act by blocking leukocyte penetration of the blood brain barrier. However, in view of unsatisfactory results and severe side effects, complementary therapies are needed. We have examined the effect of chlorite-oxidized oxyamylose (COAM, a potent antiviral polycarboxylic acid on EAE. Methods EAE was induced in SJL/J mice by immunization with spinal cord homogenate (SCH or in IFN-γ-deficient BALB/c (KO mice with myelin oligodendrocyte glycoprotein peptide (MOG35-55. Mice were treated intraperitoneally (i.p. with COAM or saline at different time points after immunization. Clinical disease and histopathology were compared between both groups. IFN expression was analyzed in COAM-treated MEF cell cultures and in sera and peritoneal fluids of COAM-treated animals by quantitative PCR, ELISA and a bioassay on L929 cells. Populations of immune cell subsets in the periphery and the central nervous system (CNS were quantified at different stages of disease development by flow cytometry and differential cell count analysis. Expression levels of selected chemokine genes in the CNS were determined by quantitative PCR. Results We discovered that COAM (2 mg i.p. per mouse on days 0 and 7 protects significantly against hyperacute SCH-induced EAE in SJL/J mice and MOG35-55-induced EAE in IFN-γ KO mice. COAM deviated leukocyte trafficking from the CNS into the periphery. In the CNS, COAM reduced four-fold the expression levels of the neutrophil CXC chemokines KC/CXCL1 and MIP-2/CXCL2. Whereas the effects of COAM on circulating blood and splenic leukocytes were limited, significant

  19. 血液中白细胞CD18和CD54与急性脑梗死脑组织损伤的相关性分析%Correlated analysis of the leukocyte CD18 and CD54 of blood in patients with acute cerebral infarction

    王苏平; 张志清; 栾桂红; 赵耿毅; 姜宏彦


    @@ Background: Recently the relationship between cell adhesion molecule (CAM ) and cerebral arterial thrombosis are more and more interested. With the data of the animal experiment becoming more and more plentive, there has been some significant findings. But there is not much research on patients. Now in this article we will detect the expression of CD18 and CD54 in blood leukocyte of 88 patients with ACI by using flow cytometer and explore its clinical significance.

  20. Repopulation of lymph nodes of dogs after 1200 R whole-body x-irradiation and intravenous administration of mononuclear blood leukocytes

    Fresh and cryopreserved autologous or allogeneic mononuclear blood cells (MBCs) intravenously injected in 1200 R total-body x-irradiated dogs repopulated lymph nodes within 10 days after transfusion. Several parameters of the lymphopoietic regeneration were correlated with the number of cells transfused and with the number of colony-forming units contained in the cell suspension when they were cultured in agar (CFU/sub c/). Values within the normal or close to normal range were reached in the mesenteric nodes of most of the animals transfused with 10 x 109 MBC or more. These values were obtained when 5 x 105 CFU/sub c/ or more were transfused. Axillary nodes showed lower values than mesenteric nodes. They were mostly under the normal range but well over those of the irradiated controls. Frozen and thawed MBCs seem to be as effective as fresh cells for lymphopoietic restoration. The mesenteric nodes of dogs transfused with allogeneic MBCs showed higher cellularity and larger cortical-paracortical areas than those of dogs transfused with approximately the same number of autologous cells. The repopulation of lymph nodes parallels that of the marrow

  1. Evaluation of the Combined Effects of Gamma Radiation and High Dietary Iron on Peripheral Leukocyte Distribution and Function

    Crucian, Brian E.; Morgan, Jennifer L. L.; Quiriarte, Heather A.; Sams, Clarence F.; Smith, Scott M.; Zwart, Sara R.


    NASA is concerned with the health risks to astronauts, particularly those risks related to radiation exposure. Both radiation and increased iron stores can independently increase oxidative damage, resulting in protein, lipid and DNA oxidation. Oxidative stress increases the risk of many health problems including cancer, cataracts, and heart disease. This study, a subset of a larger interdisciplinary investigation of the combined effect of iron overload on sensitivity to radiation injury, monitored immune parameters in the peripheral blood of rats subjected to gamma radiation, high dietary iron or both. Specific immune measures consisted of (A) peripheral leukocyte distribution; (B) plasma cytokine levels; (C) cytokine production profiles following whole blood stimulation of either T cells or monocytes.

  2. 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

    贾宇臣; 其其格; 郭菊红; 赵海珍; 乌云; 奥敦托娅


    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

  3. Prediction based on mean subset

    Øjelund, Henrik; Brown, P. J.; Madsen, Henrik;


    the coefficient vectors from each subset should be weighted. It is not computationally feasible to calculate the mean subset coefficient vector for larger problems, and thus we suggest an algorithm to find an approximation to the mean subset coefficient vector. In a comprehensive Monte Carlo simulation study...

  4. Computational modeling of leukocyte adhesion cascade (LAC)

    Sarkar, Kausik


    In response to an inflammation in the body, leukocytes (white blood cell) interact with the endothelium (interior wall of blood vessel) through a series of steps--capture, rolling, adhesion and transmigration--critical for proper functioning of the immune system. We are numerically simulating this process using a Front-tracking finite-difference method. The viscoelastcity of the cell membrane, cytoplasm and nucleus are incorporated and allowed to change with time in response to the cell surface molecular chemistry. The molecular level forces due to specific ligand-receptor interactions are accounted for by stochastic spring-peeling model. Even though leukocyte rolling has been investigated through various models, the transitioning through subsequent steps, specifically firm adhesion and transmigration through endothelial layer, has not been modeled. The change of viscoelastic properties due to the leukocyte activation is observed to play a critical role in mediating the transition from rolling to transmigration. We will provide details of our approach and discuss preliminary results.

  5. Mechanisms of leukocyte transendothelial and interstitial migration during inflammation in vivo

    Khandoga, Alexander


    The emigration of leukocytes from the circulation is a critical step during immune surveillance and inflammatory reactions that is governed by a coordinated interplay involving a spectrum of adhesion and signal molecules. While a great deal has been learned about the early steps of leukocyte recruitment, i.e. rolling and adhesion, little is known about the subsequent steps, transendothelial and interstitial migration when leukocytes migrate across the endothelial layer lining the blood vessel...

  6. Quality control in a manual and an automated leukocyte differential count.

    Takubo, T; Tatsumi, N


    Quality control (QC) has been introduced in laboratories, and QC surveys in leukocyte differential count to enhance quality have been performed by College of American Pathologists, Japanese Association of Medical Technologists, Osaka Medical Association and manufacturers. The results of QC survey in a manual leukocyte differential count indicated problems on the differentiation of segmented neutrophils and band neutrophils and the detection of pathological blood cells on blood smear. While the results of QC survey in an automated leukocyte differential count performed by same manufacturer with an automated blood cell counter were satisfactory, however, there was a difference in leukocyte differential cell counts among laboratories with other manufacturer's instruments because the synthetic blood material used in QC is an exclusive item for an instrument. It is necessary to further reeducate the medical technologists in order to improve morphological performance, and to standardize the synthetic blood material for compatibility with various automated blood cell counters. PMID:10926263

  7. 危重症手足口病患儿淋巴细胞亚群的检测%Analysis on peripheral blood lymphocyte subsets in children with critical hand-foot-mouse disease

    莫扬; 李智山


    目的 研究危重症手足口病患儿外周血淋巴细胞亚群的变化,以探讨其免疫功能改变.方法 以临床确诊并入住ICU的40例危重症手足口病患儿为研究对象,并与30例正常儿童进行比对研究,应用流式细胞仪检测外周抗凝全血的淋巴细胞亚群:T淋巴细胞(CD3+CD19-)、辅助T细胞Th(CD3+CD4+)、抑制T细胞Ts(CD3+CD8+)、B细胞(CD3-CD19+)和NK细胞(CD3-CD56+)的相对计数,并计算Th/Ts值.结果 危重症手足口病患儿外周血T淋巴细胞、Th细胞、Ts细胞的百分率下降,均明显低于对照组(P<0.05);B细胞的百分率则显著增高(P<0.05);而两组间NK细胞百分率及Th/Ts细胞比值的变化不存在明显差异(P>0.05).结论 危重症手足口病患儿出现细胞免疫及体液免疫功能紊乱.%Objective To study the variation of perpheral blood lymphocyte subsets and inmune function in critical hand-fcotr-mouse disease (HFMD ) children .M ethods Forly cases with critical HFMD and thirty healthy children as control weie enrolled in the study .The percentage of lym phocyte subsets,inclidhg CD3+CD19-cells(T),CD3+CD4+cells(Th) ,CD3+CD8+cells(Ts), CD3-eD19+cells(B),CD3-eD56+cells(NK ),in anticoagulative whole bbod of patients Son 1he too groupsweie detemined by fkw cyfcm etiy .Results Ccmpaied with nomal controls ,1he Critical HFMD children showed an obvious decrease in percentage of CD3+CD19-cells,T helper cells ,T suppressor cells,but obvious increase ofCD3-CD 19+ cells .No significant difieiencew as found in the ratio ofTh/Ts and CD 3-CD 56+cells bete een the too groups .Conclusion There aie functional disoiders in cellular and hum oral inmunity in criticalHFMD children .

  8. Factors affecting leukocyte count in healthy adults.

    Carel, R S; Eviatar, J


    The relationships between white blood cell (WBC) count, smoking, and other health variables were determined among 35,000 apparently healthy men and women. The effect of smoking on the WBC count was greater than that of all other variables. The leukocyte level and the variance in WBC count values increased with increased smoking intensity. The relationship between smoking intensity and leukocyte level is expressed quantitatively by the following regression equation: WBC (10(3)/mm3) = 7.1 + 0.05(SM), where SM has seven values according to the smoking level. Multiple regression analysis with additional variables other than smoking did not much improve the predictive value of the equation. The effect of smoking on WBC count could be only partially explained by an inflammatory process, e.g., chronic bronchitis. Relationships of statistical significance (but mostly with r values of less than 0.10) were found between WBC count and the following variables: hemoglobin, heart rate, weight (or Quetelet index), cholesterol, uric acid, creatinine, sex, ethnic origin, systolic blood pressure, height, blood sugar, and diastolic blood pressure. The normal WBC count range for smokers differs from that of nonsmokers and is shifted to the right according to the smoking level. This may have both a diagnostic and prognostic significance in different clinical settings. PMID:4070192

  9. Expression of β2-integrin on leukocytes in liver cirrhosis

    Anatol Panasiuk; Janusz Zak; Elzbieta Maciorkowska; Bozena Panasiuk; Danuta Prokopowicz


    AIM: To analyze β2-integrin expression on blood leukocytes in liver cirrhosis.METHODS: In 40 patients with liver cirrhosis and 20healthy individuals, the evaluation of expression of CD11a (LFA-1α), CD11b (Mac-1α), CD11c (αX) and CD49d (VLA-4α) on peripheral blood leukocytes was performed using flow cytometry. The analysis was carried out in groups of patients divided into B and C according to Child-Pugh's classification.RESULTS: An increased CD11a, CD11b, CD11c and CD49d integrin expression was observed on peripheral blood leukocytes in liver cirrhosis. The integrin levels were elevated as the advancement of liver failure progressed. The highest expression of integrins occurred predominantly on monocytes. A slight expression of VLA-4 was found on lymphocytes and granulocytes and it increased together with liver failure. A positive correlation was noted between median intensity of fluorescence (MIF) expression on polymorphonuclear cells of CD11a and CD11c and CD49d (r = 0.42, P < 0.01; r = 053, P < 0.01, respectively) in liver cirrhosis stage C. However,no correlation was observed between integrin expression on leukocytes. The concentrations of sICAM-1, sVCAM-1,and TNFα, were significantly elevated in liver cirrhosis.CONCLUSION: β2-integrin expression on leukocytes increases in liver cirrhosis decompensated as the stage of liver failure increases, which is a result of permanent activation of leukocytes circulating through the inflamed liver environment. β2-integrin expression on circulating leukocytes can intensify liver cirrhosis.

  10. Subset sums in abelian groups

    Balandraud, Eric; Girard, Benjamin; Griffiths, Simon; Hamidoune, Yahya Ould


    Denoting by Sigma(S) the set of subset sums of a subset S of a finite abelian group G, we prove that |Sigma(S)| >= |S|(|S|+2)/4-1 whenever S is symmetric, |G| is odd and Sigma(S) is aperiodic. Up to an additive constant of 2 this result is best possible, and we obtain the stronger (exact best possible) bound in almost all cases. We prove similar results in the case |G| is even. Our proof requires us to extend a theorem of Olson on the number of subset sums of anti-symmetric subsets S from the...

  11. Superoxide production by phagocytic leukocytes.

    Drath, D B; Karnovsky, M L


    Mononuclear phagocytic leukocytes, as well as polymorphonuclear leukocytes, produce and release superoxide at rest, and this is stimulated by phagocytosis. Of the mouse monocytic cells studied, alveolar macrophages released the largest amounts of superoxide during phagocytosis, followed by normal peritoneal macrophages. Casein-elicited and "activated" macrophages released smaller quantities. In the guinea pig, polymorphonuclear leukocytes and casein-elicited macrophages were shown to release superoxide during phagocytosis whereas alveolar macrophages did not. Superoxide release accounted for only a small fraction of the respiratory burst of phagocytosis in all but the normal mouse peritoneal macrophage, the guinea pig polymorphonuclear leukocyte, and probably the mouse alveolar macrophage. There are obviously considerable species differences in O2-release by various leukocytes that might reflect both the production and/or destruction (e.g. by dismutase) of that substance. PMID:804030

  12. Leukocyte biophysics. An invited review.

    Schmid-Schönbein, G W


    The biophysical properties of leukocytes in the passive and active state are discussed. In the passive unstressed state, leukocytes are spherical with numerous membrane folds. Passive leukocytes exhibit viscoelastic properties, and the stress is carried largely by the cell cytoplasm and the nucleus. The membrane is highly deformable in shearing and bending, but resists area expansion. Membrane tension can usually be neglected but plays a role in cases of large deformation when the membrane becomes unfolded. The constant membrane area constraint is a determinant of phagocytic capacity, spreading of cells, and passage through narrow pores. In the active state, leukocytes undergo large internal cytoplasmic deformation, pseudopod projection, and granule redistribution. Several different measurements for assessment of biophysical properties and the internal cytoplasmic deformation in form of strain and strain rate tensors are presented. The current theoretical models for active cytoplasmic motion in leukocytes are discussed in terms of specific macromolecular reactions. PMID:1705479

  13. In-111-labeled leukocyte brain SPECT imaging in acute ischemic stroke in man

    Fujinuma, Kunihiko; Sakai, Fumihiko; Iizuka, Takahiro; Kitai, Norio [Kitasato Univ., Sagamihara, Kanagawa (Japan). School of Medicine


    This study was performed to investigate the role of leukocyte accumulation in human cerebral infarction and its association with neurological functional outcome. A total of 42 patients diagnosed as acute ischemic stroke (22 embolism, 17 thrombosis, 3 TIA) were examined. Leukocyte accumulation was studied using indium-111-labeled leukocyte brain SPECT. Volume of brain infarction was evaluated by CT and/or MRI. The data were compared with the cerebral blood flow (CBF) imaging. Immediately after CBF study by SPECT using either Tc-99m-HMPAO or Tc-99m-ECD, In-111-labeled autologous leukocytes were injected intravenously. Brain scan for leukocytes was performed after 48 hours. The European Stroke Scale was used for neurological assessment. Thirteen patients with cerebral embolism and three patients with cerebral thrombosis showed intensive accumulation of leukocytes in the region of low flow Leukocyte`s accumulation was not seen in patients with TIA. The accumulation of leukocytes was more noticeable in the central zone of the ischemia. Patients who showed negative leukocyte accumulation revealed clinically mild functional outcome and the size of infarction on CT and/or MRI was small. The regional accumulation of leukocytes was seen in all the patients with hemorrhagic infarction, but the degree of hemorrhage on CT did not have significant influence on the amount of leukocyte accumulation. Abnormal accumulation of leukocytes was associated with reduced CBF during the acute embolic stroke. The present clinical study revealed that leukocyte accumulation correlated with the poor neurological functional outcome in patients with acute embolic stroke. (K.H.)

  14. Metallothionein mediates leukocyte chemotaxis

    Lynes Michael A


    Full Text Available Abstract Background Metallothionein (MT is a cysteine-rich, metal-binding protein that can be induced by a variety of agents. Modulation of MT levels has also been shown to alter specific immune functions. We have noticed that the MT genes map close to the chemokines Ccl17 and Cx3cl1. Cysteine motifs that characterize these chemokines are also found in the MT sequence suggesting that MT might also act as a chemotactic factor. Results In the experiments reported here, we show that immune cells migrate chemotactically in the presence of a gradient of MT. This response can be specifically blocked by two different monoclonal anti-MT antibodies. Exposure of cells to MT also leads to a rapid increase in F-actin content. Incubation of Jurkat T cells with cholera toxin or pertussis toxin completely abrogates the chemotactic response to MT. Thus MT may act via G-protein coupled receptors and through the cyclic AMP signaling pathway to initiate chemotaxis. Conclusion These results suggest that, under inflammatory conditions, metallothionein in the extracellular environment may support the beneficial movement of leukocytes to the site of inflammation. MT may therefore represent a "danger signal"; modifying the character of the immune response when cells sense cellular stress. Elevated metallothionein produced in the context of exposure to environmental toxicants, or as a result of chronic inflammatory disease, may alter the normal chemotactic responses that regulate leukocyte trafficking. Thus, MT synthesis may represent an important factor in immunomodulation that is associated with autoimmune disease and toxicant exposure.

  15. 库伦驴血液白细胞抗菌肽提取及其对牛乳腺炎主要致病菌的抗菌活性研究%Antibacterial activity of antibacterial peptides from Kulun Donkey blood leukocytes against the major pathogens responsible for bovine mastitis

    布日额; 吴金花; 卢雅生


    Objective To verify the in vitro activity of antibacterial peptides from Kulun Donkey blood leukocytes bovine mastitis caused by Streptococcus agalactiae, Staphylococcus aureus, and Escherichiacoli. Methods Antibacterial peptides were extracted from Kulun Donkey blood leukocytes using acetic acid extraction. Drug susceptibility test strips were prepared and antibacterial activity was determined using the agar plate method. Results Antibacterial peptides from Kulun Donkey blood leukocytes had activity against bovine mastitis caused by S. Agalactiae, S. Aureus, and E. Coli. The bacterial inhibition ring diameter was 14 mm, 15 mm, and 17mm, respectively, with 43. 60 mg/ml antibacterial peptides 24 h after incubation. Conclusion Antibacterial peptides from Kulun Donkey blood leukocytes inhibited the bacterial activity of S. Agalactiae. S. Aureus, and E. Coli. These peptides could be used to treat bovine mastitis caused by these pathogens.%目的 验证库伦驴血液白细胞抗菌肽对牛乳腺炎无乳链球菌、金黄色葡萄球菌及大肠埃希菌的体外抗菌活性.方法 利用乙酸萃取法提取库伦驴血液白细胞抗菌肽,制备药敏试纸片,用平皿药敏纸片法检测其抗菌活性. 结果 库伦驴血液白细胞抗菌肽对无乳链球菌、金黄色葡萄球菌及大肠埃希菌具有抑菌效果,43.60 mg/ml抗菌肽作用24 h抑菌环直径分别为14、15和17 mm. 结论 库伦驴白细胞抗菌肽对无乳链球菌、金黄色葡萄球菌、大肠埃希菌有抑菌活性,可用于上述致病菌感染所致奶牛乳腺炎的治疗.

  16. Leukocyte differential count in peripheral blood "with" single tube/ten colors"flow cytometry%单管10色流式细胞术分类计数外周血白细胞

    刘雪凯; 潘娜; 王建中; 邢莹; 屈晨雪; 普程伟; 伍平; 王卉; 刘抗援; 马兰


    目的 应用"单管10色"流式细胞术分类计数外周血白细胞,并探讨其临床应用价值.方法 通过多种单克隆抗体的选择和多重逻辑设门的研究,确定了10色流式细胞术与12种单克隆抗体组合的单管免洗外周血白细胞分类方法(简称"单管10色"方法).选取142份外周血标本,以血涂片显微镜法分类技术为参考方法,同时用Beckman Cotdter LH750全自动血细胞分析仪和"单管10色"方法分类计数白细胞,计算"单管10色"方法的临床诊断效率,并统计分析两种方法与显微镜法分类计数白细胞的相关性.以BD FACS Calibur流式细胞仪免疫分型结果为标准,计算"单管10色"流式细胞术检出原始细胞的诊断效率.结果 "单管10色"流式细胞术与显微镜法分类计数外周血白细胞除嗜碱粒细胞外均具有良好的相关性(r均>0.700,P均0.700,P<0.01) except for basophils.The correlations with neutrophilic granulocytes,lymphocytes,immature granulocytes and blasts were superior(r=0.972,0.951,0.801,0.912,respectively,P<0.01).When 1% was selected as the cut-off point for immature granulocytes determined by standard microscopic cytology,the sensitivity and the specificity of"single tube/10 colors"flow cytometry were 92%(57/62) and 79% (63/80),respectively.When 0.5% was selected as the cut-off point for blasts detected by standard microscopic cytology,the sensitivity and the specificity were 99% (67/68) and 92% (68/74).Using the immunophenotyping results from BD FACS Calibur as a standard,the sensitivity for detecting blasts by"single tube/10 colOrs"flow cytometry was 100% (40/40),the specificity was 91% (10/11),the positive predictive value was 98% (40/41),the negative predictive value was 100% (10/10) and the accuracy was 98% (50/51).Conclusions The"single tube/10 colors"flow cytometry has a excellent correlation with the standard microscopic cytology when applied on leukocyte differential count in peripheral blood.It may potentially

  17. Association between diabetes complications and leukocyte counts in Iranian patients

    Moradi S


    Full Text Available Sedigheh Moradi1, Scott Reza Jafarian Kerman2, Farzaneh Rohani1, Fereshteh Salari21Endocrine and Metabolism Research Center (Firouzgar, Hemmat Campus, Tehran University of Medical Sciences, Tehran, Iran, 2Scientific Students Research Committee, Tehran University of Medical Sciences, Tehran, IranBackground: The long term complications of diabetes can be fatal. They are also renowned for being an economic burden. Previous studies have demonstrated a relationship between inflammatory markers and complications of diabetes. The objective of this study was to evaluate the relationship between leukocyte counts and these complications.Methods: The study included 184 patients diagnosed with type 2 diabetes. The study was carried out in Iran during 2007 and 2008. Data collected on the subjects were as follows: age, gender, weight, height, blood pressure, smoking history, lipid profile including low density lipoprotein (LDL, high density lipoprotein (HDL, total cholesterol, triglycerides, and leukocyte count, albuminuria, and retinopathy. Furthermore, information on cardiac history for 100 patients was collected. The subjects were split into two groups according to their leukocyte levels: low (≤7000/mm³ and high (>7000/mm³; and then analyzed by Student's t-test or Mann–Whitney U-test as appropriate.Results: The average leukocyte count in these patients was 7594 ± 1965/mm³. Leukocyte count was significantly different in patients with and without retinopathy and albuminuria (P < 0.0001. According to this analysis, a leukocyte count of 6750/mm³ with a sensitivity of 80.2% and a specificity of 56.4%, and a count of 7550/mm³ with a sensitivity of 63.2% and a specificity of 74.6% indicated at least one diabetes complication.Conclusion: An elevated leukocyte count even within the normal range was associated with chronic complications in type 2 diabetes.Keywords: leukocytes, diabetes complications, inflammation

  18. Kinetics of reversible-sequestration of leukocytes by the isolated perfused rat lung

    Goliaei, B.


    The kinetics and morphology of sequestration and margination of rat leukocytes were studied using an isolated perfused and ventilated rat lung preparation. Whole rat blood, bone marrow suspension, or leukocyte suspensions, were used to perfuse the isolated rat lung. The lung was also perfused with latex particle suspensions and the passage of particles through the lung capillaries was studied. When a leukocyte suspension was perfused through the lung in the single-pass mode, the rate of sequestration decreased as more cells were perfused. In contrast, latex particles of a size comparable to that of leukocytes were totally stopped by the lung. When the leukocyte suspension was recirculated through the lung, cells were rapidly removed from circulation until a steady state was reached, after which no net removal of cells by the lung occurred. These results indicate that leukocytes are reversibly sequestered from circulation. The sequestered cells marginated and attached to the luminal surface of the endothelium of post-capillary venules and veins. A mathematical model was developed based on the assumption that the attachment and detachment of leukocytes to blood vessel walls follows first-order kinetics. The model correctly predicts the following characteristics of the system: (a) the kinetics of the sequestration of leukocytes by the lung; (b) the existence of a steady state when a suspension of leukocytes is recirculated through the lung; and (c) the independence of the fraction of cells remaining in circulation from the starting concentration for all values of starting concentration. (ERB)

  19. Leukocyte Activation in Obese Patients

    Minervino, Daniele; Gumiero, Daniela; Nicolazzi, Maria Anna; Carnicelli, Annamaria; Fuorlo, Mariella; Guidone, Caterina; Di Gennaro, Leonardo; Fattorossi, Andrea; Mingrone, Geltrude; Landolfi, Raffaele


    Abstract The rising prevalence of obesity is a major global health problem. In severe obesity, bariatric surgery (BS) allows to obtain a significant weight loss and comorbidities improvement, among them one of the factors is the thrombotic risk. In this observational study, we measured indices of leukocyte activation in severely obese patients as markers of increased thrombotic risk in relation with serum markers of inflammation before and after BS. Frequency of polymorphonuclear neutrophil-platelet (PLT) and monocyte (MONO)-PLT aggregates as well as of tissue factor (TF) expressing MONOs was measured in the peripheral blood of 58 consecutive obese patients and 30 healthy controls. In 31 of the 58 obese patients, data obtained at the enrollment were compared with those obtained at 3, 6, and 12 months after BS. Compared with healthy controls, obese patients showed a higher frequency of polymorphonuclear leukocyte (PMNL)-PLT aggregates (7.47 ± 2.45 [6.82–8.11]% vs 5.85 ± 1.89 [5.14–6.55]%, P = 0.001), MONO-PLT aggregates (12.31 ± 7.33 [10.38–14.24]% vs 8.14 ± 2.22 [7.31–8.97]%, P < 0.001), and TF expressing MONOs (4.01 ± 2.11 [3.45–4.56]% vs 2.64 ± 1.65 [2.02–3.25]%, P = 0.002). PMNL-PLT and MONO-PLT aggregate frequency was positively correlated with TF expressing MONOs (R2 = 0.260, P = 0.049 and R2 = 0.318, P = 0.015, respectively). BS was performed in 31 patients and induced a significant reduction of the body mass index, and waist and hip circumferences. These effects were associated with a significant decrease of PMNL-PLT aggregates at 12 months (7.58 ± 2.27 [6.75–8.42]% vs 4.47 ± 1.11 [3.93–5.01]%, P < 0.001), and a reduction of TF expressing MONOs at 6 (3.82 ± 2.04 [3.07–4.57]% vs 1.60 ± 1.69 [0.30–2.90]%, P = 0.008) and 12 months (3.82 ± 2.04 [3.07–4.57]% vs 1.71 ± 0.54 [1.45–1.97]%, P = 0.001) after BS. These data suggest that leukocyte

  20. Reference ranges of hematology and lymphocyte subsets in healthy Korean native cattle (Hanwoo) and Holstein dairy cattle.

    Kim, Yun-Mi; Lee, Jin-A; Jung, Bock-Gie; Kim, Tae-Hoon; Lee, Bong-Joo; Suh, Guk-Hyun


    There are no accurate reference ranges for hematology parameters and lymphocyte subsets in Korean native beef cattle (Hanwoo). This study was performed to establish reliable reference ranges of hematology and lymphocyte subsets using a large number of Hanwoo cattle (n = 350) and to compare differences between Hanwoo and Holstein dairy cattle (n = 334). Additionally, age-related changes in lymphocyte subsets were studied. Bovine leukocyte subpopulation analysis was performed using mono or dual color flow cytometry. The leukocyte subpopulations investigated in healthy cattle included: CD2(+) cells, sIgM(+) cells, MHC class II(+) cells, CD3(+) CD4(+) cells, CD3(+) CD8(+) cells, and WC1(+) cells. Although Hanwoo and Holstein cattle are the same species, results showed several differences in hematology and lymphocyte subsets between Hanwoo and Holstein cattle. This study is the first report to establish reference ranges of hematology and lymphocyte subsets in adult Hanwoo cattle. PMID:26419947

  1. Efficient Feature Subset Selection and Subset Size Optimization

    Somol, Petr; Novovičová, Jana; Pudil, Pavel

    Vukovar, Croatia: In-Teh, 2010 - (Herout, A.), s. 75-98 ISBN 978-953-7619-90-9 R&D Projects: GA MŠk 1M0572; GA ČR GA102/08/0593; GA ČR GA102/07/1594 Grant ostatní: GA MŠk(CZ) 2C06019 Institutional research plan: CEZ:AV0Z10750506 Keywords : dimensionality reduction * pattern recognition * machine learning * feature selection * optimization * subset search * classification Subject RIV: BD - Theory of Information feature subset selection and subset size optimization.pdf

  2. Scintigraphic differential diagnosis in painful hip endoprothesis with labelled leukocytes

    In patients with painful prosthetic joints of the hip scintigraphy with white blood cells is a useful method to differentiate between loosening and infection. Examinations of 34 patients after labelling of autologous leukocytes with 111In-oxinate or with 99mTc-DPO gave good results, involving decisive advantages (availability, acquisition time, costs) of 99mTc-DPO. (author)

  3. Leukocyte telomere length predicts overall survival in hepatocellular carcinoma treated with transarterial chemoembolization.

    Liu, Han-Qiang; An, Jia-Ze; Liu, Juan; Yang, Ye-Fa; Zhang, Hong-Xin; Zhao, Bin-Yu; Li, Ji-Bin; Yang, Hu-Shan; Chen, Zhi-Nan; Xing, Jin-Liang


    Previous studies have reported that telomere length in peripheral blood leukocytes can predict the clinical outcome of several cancers. However, whether leukocyte telomere length is associated with the prognosis of hepatocellular carcinoma (HCC) remains to be determined. In this study, relative telomere length (RTL) in peripheral blood leukocytes was measured using a real-time PCR-based method for 269 HCC patients treated with transarterial chemoembolization (TACE) from two independent hospitals. The association between RTL and the overall survival (OS) of HCC was analyzed. The immunological function of the HCC patients with different leukocyte RTLs was evaluated. Multivariate analyses indicated that long leukocyte RTL was significantly associated with poor OS of HCC patients, with a hazard ratio of 2.04 (95% confidence interval, 1.46-2.86; P HCC patients treated with TACE. PMID:22318909

  4. In-111-labeled leukocyte brain SPECT imaging in acute ischemic stroke in man

    This study was performed to investigate the role of leukocyte accumulation in human cerebral infarction and its association with neurological functional outcome. A total of 42 patients diagnosed as acute ischemic stroke (22 embolism, 17 thrombosis, 3 TIA) were examined. Leukocyte accumulation was studied using indium-111-labeled leukocyte brain SPECT. Volume of brain infarction was evaluated by CT and/or MRI. The data were compared with the cerebral blood flow (CBF) imaging. Immediately after CBF study by SPECT using either Tc-99m-HMPAO or Tc-99m-ECD, In-111-labeled autologous leukocytes were injected intravenously. Brain scan for leukocytes was performed after 48 hours. The European Stroke Scale was used for neurological assessment. Thirteen patients with cerebral embolism and three patients with cerebral thrombosis showed intensive accumulation of leukocytes in the region of low flow Leukocyte's accumulation was not seen in patients with TIA. The accumulation of leukocytes was more noticeable in the central zone of the ischemia. Patients who showed negative leukocyte accumulation revealed clinically mild functional outcome and the size of infarction on CT and/or MRI was small. The regional accumulation of leukocytes was seen in all the patients with hemorrhagic infarction, but the degree of hemorrhage on CT did not have significant influence on the amount of leukocyte accumulation. Abnormal accumulation of leukocytes was associated with reduced CBF during the acute embolic stroke. The present clinical study revealed that leukocyte accumulation correlated with the poor neurological functional outcome in patients with acute embolic stroke. (K.H.)

  5. Labeled polymorphonuclear leukocytes: a comparison of methodology

    Polymorphonuclear neutrophilic granulocytes were separated from anti-coagulated whole blood using three techniques. The methods employed included volex sedimentation (VS), volex sedimentation with hypotonic lysis (VSHL), and Ficoll-Hypaque gradient separation (FH). The cells were labeled with 111In-oxine and 111In-tropolone. Studies were done with both blood from normal human volunteers and with canine blood. From the cell counts and differential, the harvested granulocytes, platelets, and red blood cells per milliliter of whole blood were calculated. Using the granulocyte chemotactic response to E. coli in agarose plates, the ratio of chemotactic migration to random migration (c./r.m.) was determined. Survival time for 111In labeled granulocytes were also determined in a canine model. The studies demonstrated that all procedures yielded 100% viability by the Trypan blue exclusion test. Chemotactic migration and leukocyte survival times were similar amongst all techniques. With the VSHL technique, there were significantly fewer red blood cells and platelets in the final preparation approaching the results of FH separation. The results suggest that, for a relatively pure granulocyte preparation, VSHL is an acceptable alternative to FH. (author)

  6. Four-part differential leukocyte count using μflow cytometer

    Shi, Wendian; Kasdan, Harvey L.; Fridge, Alan; Tai, Yu-Chong


    This paper reports the four-part differential leukocyte count (DLC) of human blood using a MEMS microflow (μflow) cytometer. It is achieved with a two-color laser-induced fluorescence (LIF) detection scheme. Four types of leukocytes including neutrophils, eosinophils, lymphocytes and monocytes are identified in blood samples, which are stained by fluorescein isothiocyanate (FITC) and propidium iodide (PI). The DLC results show good correlation with the count from a ...

  7. Clinical application of Tc-99m HMPAO labeled leukocyte imaging in inflammatory disease

    A radionuclide imaging with Tc-99m HMPAO labeled leukocyte was performed in order to determine its clinical usefulness in inflammatory disease. The mixed leukocyte isolated from 40 ml of whole blood containing 5 ml of acid citrate dextrose A and 7 ml of 6% hydroxyethyl starch was incubated with 370 MBq (10 mCi) of Tc-99m HMPAO at 37degC for 30 minutes. Because the labeling efficiency of Tc-99m HMPAO labeled leukocyte was 60.2±6.3%, the procedure of washing Tc-99m leukocyte with 5 ml of physiological saline was necessary before intravenous injection, in order to remove the unlabeled Tc-99m HMPAO. The recoveries of Tc-99m leukocyte in the blood after intravenous injection were 41.1±6.7% at 5 minutes, 33.4±2.1% at 30 minutes, and 27.2±3.4% at 2 hours after injection. Moreover, the labeled leukocyte was not stained with trypan blue. Therefore, the biological activity of the Tc-99m leukocyte was maintained as that of In-111 oxine labeled leukocyte. In the 39 patients with clinical suspicion of inflammatory disease including 15 patients with acute and chronic infectious disease where both Tc-99m leukocyte and Ga-67 citrate imagings were performed, the sensitivity, specificity and accuracy for infectious disease were 47%, 100%, and 79% with Tc-99m leukocyte, and 67%, 79%, 74% with Ga-67 citrate. These results suggest that Tc-99m HMPAO labeled leukocyte imaging is promising for evaluating inflammatory disease because of much higher specificity, the ready availability of Tc-99m HMPAO, the good image quality, and the lower radiation dose to the patient. (author)

  8. Quantitation of acute experimental ocular inflammation with 111indium-leukocytes

    The cellular component of an acute ocular inflammation in rabbits was measured with autologous leukocytes exogenously labeled with 111Indium tropolonate. Inflammation was induced by intravitreal bacterial lipopolysaccharide (LPS). After 16 hr blood was removed, leukocytes separated, labeled with 111Indium tropolonate and reinjected. Three cell fractions were examined: a leukocyte rich fraction which had been prepared with Dextran; and polymorphonuclear and mononuclear leukocyte fractions which had been prepared using a discontinuous Percoll gradient. Two hours after labeled leukocytes were injected, measurements of 111Indium were made in blood, plasma, the whole eye and in ocular compartments. From these data the numbers of each leukocyte population present were estimated and compared directly to histopathologic changes. Both polymorphonuclear and mononuclear leukocytes entered ocular tissues during the 2 hr period beginning 20 hr after LPS injection. Altered ocular vascular permeability was successfully measured with 125Iodine-albumin in some of these same rabbits. Both the number and type of inflammatory cell entering ocular tissues during a set period of time of the inflammatory response could thus be measured. This technique provides an opportunity to define the relationship of leukocyte infiltration and altered ocular vascular permeability in ocular tissues during the inflammatory response

  9. Imaging leukocytes in vivo with third harmonic generation microscopy

    Tsai, Cheng-Kun; Chen, Chien-Kuo; Chen, Yu-Shing; Wu, Pei-Chun; Hsieh, Tsung-Yuan; Liu, Han-Wen; Yeh, Chiou-Yueh; Lin, Win-Li; Chia, Jean-San; Liu, Tzu-Ming


    Without a labeling, we demonstrated that lipid granules in leukocytes have distinctive third harmonic generation (THG) contrast. Excited by a 1230nm femtosecond laser, THG signals were generated at a significantly higher level in neutrophils than other mononuclear cells, whereas signals in agranular lymphocytes were one order smaller. These characteristic THG features can also be observed in vivo to trace the newly recruited leukocytes following lipopolysaccharide (LPS) challenge. Furthermore, using video-rate THG microscopy, we also captured images of blood cells in human capillaries. Quite different from red-blood-cells, every now and then, round and granule rich blood cells with strong THG contrast appeared in circulation. The corresponding volume densities in blood, evaluated from their frequencies of appearance and the velocity of circulation, fall within the physiological range of human white blood cell counts. These results suggested that labeling-free THG imaging may provide timely tracing of leukocyte movement and hematology inspection without disturbing the normal cellular or physiological status.

  10. The effects of medium-strength electric impulses on human blood.

    Filipic, B; Kovács, K; Somogyvári, F; Ihan, A; Ocsovszky, I; Koren, S; Tóth, S


    Leukocyte subsets, total leukocyte isolates or full blood samples were subjected to medium-strength square-wave electric impulses (100 V/cm field force, 5 ms duration). On the surface of the leukocytes, the expressions of several markers (CD3, CD4, CD8, CD11a, CD11b and ICAM-1) were determined in order to study the influence of pulsed ionic currents on different aspects of the cellular immune response. Large individual differences were observed among randomly chosen healthy donors, both in the initial expression rate and in the response patterns of different antigens. As a general conclusion, it can be stated that electric impulses with the above parameters activate the state of immune response alertness of human leukocytes. Changes in the activities of several enzymes in the serum in response to electric impulses were also tested in order to examine the feasibility of ex vivo electric treatment of human blood for the establishment of an antiviral and immune activated condition. Slightly elevated lactate dehydrogenase (LDH) levels point to a possibility of enhanced haemolysis, while the lack of an elevation in the membrane-bound peroxidase activity indicates the absence of haemolysis. Significant rises were detected in the serum superoxide dismutase (SOD) activities. Since most ex vivo blood manipulations are characterised by the appearance of superoxide radicals in the serum, a SOD activity enhancement is considered beneficial in these cases. A mild, but significant reduction in the blood clotting time indicates that electric treatment of human blood should be performed with special attention to thrombosis-prone conditions, and adequate precautions and countermeasures should be introduced. Although wider examinations are required before this method can be fully recommended, ex vivo blood treatment with medium-strength electric impulses seems to be a promising adjuvant course for the establishment of acute immune potentiation and an antiviral state in patients

  11. Microfluidic Leukocyte Isolation for Gene Expression Analysis in Critically Ill Hospitalized Patients

    Russom, Aman; Sethu, Palaniappan; Irimia, Daniel; Mindrinos, Michael N.; Calvano, Steve E.; Garcia, Iris; Finnerty, Celeste; Tannahill, Cynthia; Abouhamze, Amer; Wilhelmy, Julie; López, M. Cecilia; Baker, Henry V.; Herndon, David N.; Lowry, Stephen F.; Maier, Ronald V.; Davis, Ronald W.; Moldawer, Lyle L.; Tompkins, Ronald G.; Toner, Mehmet


    BACKGROUND Microarray technology is becoming a powerful tool for diagnostic, therapeutic, and prognostic applications. There is at present no consensus regarding the optimal technique to isolate nucleic acids from blood leukocyte populations for subsequent expression analyses. Current collection and processing techniques pose significant challenges in the clinical setting. Here, we report the clinical validation of a novel microfluidic leukocyte nucleic acid isolation technique for gene expression analysis from critically ill, hospitalized patients that can be readily used on small volumes of blood. METHODS We processed whole blood from hospitalized patients after burn injury and severe blunt trauma according to the microfluidic and standard macroscale leukocyte isolation protocol. Side-by-side comparison of RNA quantity, quality, and genome-wide expression patterns was used to clinically validate the microfluidic technique. RESULTS When the microfluidic protocol was used for processing, sufficient amounts of total RNA were obtained for genome-wide expression analysis from 0.5 mL whole blood. We found that the leukocyte expression patterns from samples processed using the 2 protocols were concordant, and there was less variability introduced as a result of harvesting method than there existed between individuals. CONCLUSIONS The novel microfluidic approach achieves leukocyte isolation in <25 min, and the quality of nucleic acids and genome expression analysis is equivalent to or surpasses that obtained from macroscale approaches. Microfluidics can significantly improve the isolation of blood leukocytes for genomic analyses in the clinical setting. PMID:18375483

  12. Kinetics of leukocyte sequestration in the lungs of acutely septic primates: A study using 111In-labeled autologous leukocytes

    To further clarify the role of leukocytes in the pathogenesis of ARDS, we studied the localization and kinetics of leukocyte migration using 111In-labeled autologous white cell scans (111In wbc scans) in four primates made acutely septic with infusions of Escherichia coli. Whole body images were obtained with a gamma camera and were acquired on computer every 15 min beginning immediately after the E. coli infusion. Simultaneous measurements of C5a and peripheral blood leukocyte count were also obtained. Within 5 min of initiating sepsis, three major events occurred: complement activation as measured by the production of C5a, a profound fall in peripheral leukocyte count, and a significant increase in the sequestration of leukocytes in the lungs. The pulmonary sequestration reached a peak at 15 min with a mean of 152% of baseline activity. This sequestration consisted of a population that was predominantly neutrophils. Damage to the pulmonary capillary endothelium was demonstrated by an increase in extravascular lung water. The results support a role for neutrophils and complement as mediators in the pathogenesis of ARDS

  13. Functional characterization of human Cd33+ And Cd11b+ myeloid-derived suppressor cell subsets induced from peripheral blood mononuclear cells co-cultured with a diverse set of human tumor cell lines

    Arger Nicholas


    Full Text Available Abstract Background Tumor immune tolerance can derive from the recruitment of suppressor cell populations, including myeloid-derived suppressor cells (MDSC. In cancer patients, MDSC accumulation correlates with increased tumor burden, but the mechanisms of MDSC induction remain poorly understood. Methods This study examined the ability of human tumor cell lines to induce MDSC from healthy donor PBMC using in vitro co-culture methods. These human MDSC were then characterized for morphology, phenotype, gene expression, and function. Results Of over 100 tumor cell lines examined, 45 generated canonical CD33+HLA-DRlowLineage- MDSC, with high frequency of induction by cervical, ovarian, colorectal, renal cell, and head and neck carcinoma cell lines. CD33+ MDSC could be induced by cancer cell lines from all tumor types with the notable exception of those derived from breast cancer (0/9, regardless of hormone and HER2 status. Upon further examination, these and others with infrequent CD33+ MDSC generation were found to induce a second subset characterized as CD11b+CD33lowHLA-DRlowLineage-. Gene and protein expression, antibody neutralization, and cytokine-induction studies determined that the induction of CD33+ MDSC depended upon over-expression of IL-1β, IL-6, TNFα, VEGF, and GM-CSF, while CD11b+ MDSC induction correlated with over-expression of FLT3L and TGFβ. Morphologically, both CD33+ and CD11b+ MDSC subsets appeared as immature myeloid cells and had significantly up-regulated expression of iNOS, NADPH oxidase, and arginase-1 genes. Furthermore, increased expression of transcription factors HIF1α, STAT3, and C/EBPβ distinguished MDSC from normal counterparts. Conclusions These studies demonstrate the universal nature of MDSC induction by human solid tumors and characterize two distinct MDSC subsets: CD33+HLA-DRlowHIF1α+/STAT3+ and CD11b+HLA-DRlowC/EBPβ+, which should enable the development of novel diagnostic and therapeutic reagents for

  14. A community study of T lymphocyte subsets and malaria parasitaemia

    Lisse, I M; Aaby, P; Whittle, H;


    In a community survey of 312 children aged 3-6 years in urban Guinea-Bissau, we examined Plasmodium falciparum parasitaemia and T cell subsets. 183 children (59%) had parasites in their blood, 13 had fever > or = 37.5 degrees C, and 9 (3%) had fever and a parasite density > 5000/microL (clinical ...

  15. T-lymphocyte subsets, thymic size and breastfeeding in infancy

    Jeppesen, Dorthe Lisbeth; Hasselbalch, Helle; Lisse, Ida M;


    We followed the changes in concentration of T-lymphocyte subsets (CD4+ and CD8+ cells) in peripheral blood and thymus size during infancy. Previous studies have found increased thymus size in breastfed infants. The present study analyzed the association between breastfeeding and the number of CD4...

  16. Cannabinoid-receptor expression in human leukocytes.

    Bouaboula, M; Rinaldi, M; Carayon, P; Carillon, C; Delpech, B; Shire, D; Le Fur, G; Casellas, P


    Marijuana and many of its constituent cannabinoids influence the central nervous system (CNS), probably through the cannabinoid receptor, which has recently been cloned in rat and human. While numerous reports have also described effects of cannabinoids on the immune system, the observation of both mRNA and cannabinoid receptor has hitherto been exclusively confined to the brain, a reported detection in the testis being the sole example of its presence at the periphery. Here we report the expression of the cannabinoid receptor on human immune tissues using a highly sensitive polymerase-chain-reaction-based method for mRNA quantification. We show that, although present in a much lower abundance than in brain, cannabinoid receptor transcripts are found in human spleen, tonsils and peripheral blood leukocytes. The distribution pattern displays important variations of the mRNA level for the cannabinoid receptor among the main human blood cell subpopulations. The rank order of mRNA levels in these cells is B cells > natural killer cells > or = polymorphonuclear neutrophils > or = T8 cells > monocytes > T4 cells. Cannabinoid-receptor mRNA, which is also found in monocytic, as well as T and B leukemia cell lines but not in Jurkat cells, presents a great diversity of expression on these cells as well, B-cell lines expressing a much higher level than T-cell lines. The cannabinoid receptor PCR products from leukocytes and brain are identical both in size and sequence suggesting a strong similarity between central and peripheral cannabinoid receptors. The expression of this receptor was demonstrated on membranes of the myelomonocytic U937 cells using the synthetic cannabinoid [3H]CP-55940 as ligand. The Kd determined from Scatchard analysis was 0.1 nM and the Bmax for membranes was 525 fmol/mg protein. The demonstration of cannabinoid-receptor expression at both mRNA and protein levels on human leukocytes provides a molecular basis for cannabinoid action on these cells. PMID

  17. Methamphetamine Administration Targets Multiple Immune Subsets and Induces Phenotypic Alterations Suggestive of Immunosuppression

    Robert Harms; Brenda Morsey; Craig W Boyer; Fox, Howard S.; Nora Sarvetnick


    Methamphetamine (Meth) is a widely abused stimulant and its users are at increased risk for multiple infectious diseases. To determine the impact of meth on the immune system, we utilized a murine model that simulates the process of meth consumption in a typical addict. Our phenotypic analysis of leukocytes from this dose escalation model revealed that meth affected key immune subsets. Meth administration led to a decrease in abundance of natural killer (NK) cells and the remaining NK cells p...

  18. Oligonol Supplementation Affects Leukocyte and Immune Cell Counts after Heat Loading in Humans

    Jeong Beom Lee


    Full Text Available Oligonol is a low-molecular-weight form of polyphenol and has antioxidant and anti-inflammatory activity, making it a potential promoter of immunity. This study investigates the effects of oligonol supplementation on leukocyte and immune cell counts after heat loading in 19 healthy male volunteers. The participants took a daily dose of 200 mg oligonol or a placebo for 1 week. After a 2-week washout period, the subjects were switched to the other study arm. After each supplement, half-body immersion into hot water was made, and blood was collected. Then, complete and differential blood counts were performed. Flow cytometry was used to enumerate and phenotype lymphocyte subsets. Serum concentrations of interleukin (IL-1β and IL-6 in blood samples were analyzed. Lymphocyte subpopulation variables included counts of total T cells, B cells, and natural killer (NK cells. Oligonol intake attenuated elevations in IL-1β (an 11.1-fold change vs. a 13.9-fold change immediately after heating; a 12.0-fold change vs. a 12.6-fold change 1h after heating and IL-6 (an 8.6-fold change vs. a 9.9-fold change immediately after heating; a 9.1-fold change vs. a 10.5-fold change 1h after heating immediately and 1 h after heating in comparison to those in the placebo group. Oligonol supplementation led to significantly higher numbers of leukocytes (a 30.0% change vs. a 21.5% change immediately after heating; a 13.5% change vs. a 3.5% change 1h after heating and lymphocytes (a 47.3% change vs. a 39.3% change immediately after heating; a 19.08% change vs. a 2.1% change 1h after heating relative to those in the placebo group. Oligonol intake led to larger increases in T cells, B cells, and NK cells at rest (p < 0.05, p < 0.05, and p < 0.001, respectively and immediately after heating (p < 0.001 in comparison to those in the placebo group. In addition, levels of T cells (p < 0.001 and B cells (p < 0.001 were significantly higher 1 h after heating in comparison to those in

  19. Noncytotoxic T cell clones obtained from a human mixed leukocyte culture.

    Chu, M H; Wee, S L; Bach, F H


    Peripheral blood mononuclear cells from a DQW-1 homozygous individual were cocultured with irradiated lymphoblastoid cell line from a DQW-1 homozygous unrelated donor bearing BW35-DW1 haplotype. From T cell cloning of primary and twice-stimulated mixed leukocyte cultures (MLC), 7 and 11 T cell clones were obtained respectively. None of the 18 clones showed specific cytotoxic activity against the alloantigen of the stimulator cell as well as natural killer (NK)-like activity against K562 cells. However, most T cell clones from both primary and re-stimulated MLC demonstrated moderate cytotoxic activity in lectin-dependent cell-mediated cytolysis (LDCC) assay. Screening assay for cell-mediated lympholysis (CML) performed on growing microcultures obtained from restimulated MLC cloning confirmed the non-cytotoxic status of these T cell clones by showing that 41 out of 44 growing microcultures were not cytotoxic against the stimulator cell; the other 3 clones lyzed the target cell mildly. The cells from all 5 T cell clones detected for indirect fluorescence expressed CD3 and CD4 surface markers. Taken together, the results suggested that proliferation-regulating T cell subsets or factor(s) may be generated during the course of MLCs under the present responder-stimulator combination, and may suppress the development of alloreactive cytotoxic T cells and NK-like cells. PMID:2144231

  20. Lymphocytic subsets and low-dose exposure

    The present investigations proved the differential radiosensitivity of lymphocytic subpopulations: From in vivo and in vitro irradiations it may be followed that the most sensitive subset are CD8 positive suppressor T cells. CD4/CD8 ratios are increased both in peripheral blood and after mitogen stimulation of lymphocytes of exposed persons. The decrease in B cells is pronounced only at higher radiation doses. Though the rate of DNA synthesis after mitogen stimulation was reduced in some exposed persons, that was no general phenomenon. Especially after tritium exposure, the observed lymphopenia correlated with an increased stimulation by PHA and an increased rate of DNA synthesis in some probands. Thus the present investigations indicate that - despite an inhibition of some immune parameters by radioexposure - the body is able to maintain its immunological homoeostasis. (authors)

  1. Leukocyte Responsiveness to Exercise in Individuals Positive for Human Cytomegalovirus.

    Wilson, J N; Navalta, J W


    Human cytomegalovirus (HCMV) infects 50% of adults in the United States. HCMV can become a cause for concern in individuals who have a compromised immune system, which may occur after high-intensity exercise. The purpose of this preliminary study was to characterize the lymphocyte, monocyte, and neutrophil responses to exercise in HCMV+individuals. Participants were either positive (HCMV +) or negative (HCMV-) for HCMV. Participants visited the laboratory on 3 separate occasions: HCMV screening, 100% VO2max test, and 80% VO2max run. Mixed-model factorial ANOVA procedures with repeated measures on sampling condition were performed on absolute and relative circulating lymphocytes, monocytes, and neutrophils. Significant main effects for time for both absolute and relative values were seen for all leukocyte subsets regardless of virus status. Significant differences for absolute and relative values were seen between sampling conditions for all leukocyte subsets. We report for the first time that HCMV status does not affect circulating neutrophil responses to high-intensity exercise, though exercise-induced neutrocytosis is seen during the post-exercise and 60 min post-exercise sampling conditions, regardless of HCMV status. There is no HCMV effect on circulating monocyte responses to exercise, though exercise-induced monocytosis was seen during the post-exercise sampling condition regardless of HCMV status. PMID:26837931

  2. Diagnosis of osteomyelitis of the foot in diabetic patients: Value of 111In-leukocyte scintigraphy

    The noninvasive diagnosis of osteomyelitis of the foot in diabetic patients with currently available radiologic and radionuclide imaging techniques is often difficult. Recently, 111In-labeled leukocyte scintigraphy has been proposed as an attractive alternative. Accordingly, the authors retrospectively reviewed 51 111In-labeled leukocyte scans, 49 technetium-99m bone scans, and 49 plain radiographs obtained in 51 adults with diabetes in whom osteomyelitis of the foot was suspected. The sensitivity and specificity of these techniques were evaluated in all patients, as well as in a subgroup of 11 patients with neuroarthropathy. Results with 111In-labeled leukocyte scans were also examined in subsets of patients with soft-tissue ulcers (n = 35) and those receiving antibiotics during investigation (n = 20). Confirmation or exclusion of osteomyelitis was made surgically in 28 patients and clinically in 23. Fourteen patients had osteomyelitis. Bone scans were most sensitive (93%) but least specific (43%); plain radiographs were most specific (83%) but least sensitive (43%). 111In-labeled leukocyte scans were both sensitive (79%) and specific (78%), and remained useful in patients with neuroarthropathy, soft-tissue ulcers, and antibiotic treatment. Poor spatial resolution contributed to the false-negative and false-positive 111In-labeled leukocyte scans, suggesting that this technique should not be interpreted independent of other tests. 111In-labeled leukocyte scans are a valuable diagnostic tool for the diagnosis of pedal osteomyelitis in diabetic patients

  3. Influence of rimonabant treatment on peripheral blood mononuclear cells; flow cytometry analysis and gene expression profiling

    Stefan Almestrand


    Full Text Available The cannabinoid receptor type 1 (CB1 antagonist rimonabant has been used as treatment for obesity. In addition, anti-proliferative effects on mitogen-activated leukocytes have been demonstrated in vitro. We have previously shown that rimonabant (SR141716A induces cell death in ex vivo isolated malignant lymphomas with high expression of CB1 receptors. Since CB1 targeting may be part of a future lymphoma therapy, it was of interest to investigate possible effects on peripheral blood mononuclear cells (PBMC in patients treated with rimonabant. We therefore evaluated leukocyte subsets by 6 color flow cytometry in eight patients before and at treatment with rimonabant for 4 weeks. Whole-transcript gene expression profiling in PBMC before and at 4 weeks of rimonabant treatment was done using Affymetrix Human Gene 1.0 ST Arrays. Our data show no significant changes of monocytes, B cells, total T cells or T cell subsets in PBMC during treatment with rimonabant. There was a small but significant increase in CD3–, CD16+ and/or CD56+ cells after rimonabant therapy. Gene expression analysis detected significant changes in expression of genes associated with innate immunity, cell death and metabolism. The present study shows that normal monocytes and leukocyte subsets in blood remain rather constant during rimonabant treatment. This is in contrast to the induction of cell death previously observed in CB1 expressing lymphoma cells in response to treatment with rimonabant in vitro. These differential effects observed on normal and malignant lymphoid cells warrant investigation of CB1 targeting as a potential lymphoma treatment.

  4. Improved survival of newborns receiving leukocyte transfusions for sepsis

    To determine the role of polymorphonuclear (PMN) leukocyte transfusions in neonates with sepsis, 23 consecutive newborns were prospectively randomly selected during an 18-month period in a treatment plan to receive polymorphonuclear leukocyte transfusions with supportive care or supportive care alone. Thirteen neonates received transfusions every 12 hours for a total of five transfusions. Each transfusion consisting of 15 mL/kg of polymorphonuclear leukocytes was subjected to 1,500 rads of radiation. The polymorphonuclear leukocytes were obtained by continuous-flow centrifugation leukapheresis and contained 0.5 to 1.0 X 10(9) granulocytes per 15 mL with less than 10% lymphocytes. Positive findings on blood cultures were obtained in 14/23 patients and seven were randomly selected for each treatment group. Absolute granulocyte counts were less than 1,500/microL in 13 patients but tibial bone marrow examinations revealed that the neutrophil supply pool was depleted in only three patients. The survival was significantly greater in the treatment group compared with the group that did not receive transfusions

  5. Clinical application of automated digital image analysis for morphology review of peripheral blood leukocyte%全自动数字图像分析在外周血白细胞形态学复检中的临床应用

    邢莹; 闫晓华; 普程伟; 尚柯; 董宁; 汪润; 王建中


    检查,对血细胞分析仪复检规则进行优化.结论 全自动血细胞形态学数字图像分析对外周血异常白细胞的分类计数和形态学异常检测具有较高的敏感度和特异度,可用于触发血细胞分析仪复检规则时异常白细胞的形态学复检.%Objective To explore the clinical application of automated digital image analysis in leukocyte morphology examination when review criteria of hematology analyzer are triggered.Methods The reference range of leukocyte differentiation by automated digital image analysis was established by analyzing 304 healthy blood samples from Peking University First Hospital.Six hundred and ninty-seven blood samples from Peking University First Hospital were randomly collected from November 2013 to April 2014,complete blood cells were counted on hematology analyzer,blood smears were made and stained at the same time.Blood smears were detected by automated digital image analyzer and the results were checked (reclassification) by a staff with abundant morphology experience.The same smear was examined manually by microscope.The results by manual microscopic differentiation were used as "golden standard",and diagnostic efficiency of abnormal specimens by automated digital image analysis was calculated,including sensitivity,specificity and accuracy.The difference of abnormal leukocytes detected by two different methods was analyzed in 30 samples of hematological and infectious diseases.Results Specificity of identifying abnormalities of white blood cells by automated digital image analysis was more than 90% except monocyte.Sensitivity of neutrophil toxic abnormities (including D(o)hle body,toxic granulate and vacuolization) was 100%;sensitivity of blast cells,immature granulates and atypical lymphocytes were 91.7%,60% to 81.5% and 61.5%,respectively.Sensitivity of leukocyte differential count was 91.8% for neutrophils,88.5% for lymphocytes,69.1% for monocytes,78.9% for eosinophils


    Liu Yongguo; Li Xueming; Wu Zhongfu


    The motivation of data mining is how to extract effective information from huge data in very large database. However, some redundant and irrelevant attributes, which result in low performance and high computing complexity, are included in the very large database in general.So, Feature Subset Selection (FSS) becomes one important issue in the field of data mining. In this letter, an FSS model based on the filter approach is built, which uses the simulated annealing genetic algorithm. Experimental results show that convergence and stability of this algorithm are adequately achieved.

  7. Exponentially generic subsets of groups

    Gilman, Robert; Osin, Denis


    In this paper we study the generic, i.e., typical, behavior of finitely generated subgroups of hyperbolic groups and also the generic behavior of the word problem for amenable groups. We show that a random set of elements of a nonelementary word hyperbolic group is very likely to be a set of free generators for a nicely embedded free subgroup. We also exhibit some finitely presented amenable groups for which the restriction of the word problem is unsolvable on every sufficiently large subset of words.

  8. CD31/PECAM-1 is a ligand for alpha v beta 3 integrin involved in adhesion of leukocytes to endothelium


    To protect the body efficiently from infectious organisms, leukocytes circulate as nonadherent cells in the blood and lymph, and migrate as adherent cells into tissues. Circulating leukocytes in the blood have first to adhere to and then to cross the endothelial lining. CD31/PECAM- 1 is an adhesion molecule expressed by vascular endothelial cells, platelets, monocytes, neutrophils, and naive T lymphocytes. It is a transmembrane glycoprotein of the immunoglobulin gene superfamily (IgSF), with ...


    Yabluchanskiy, A.


    Hemorrhagic stroke in response to intracerebral hemorrhage is considered an aseptic form of inflammation. With this in mind the dynamics of leukocytes changes in blood and in the hemorrhagic stroke zone as main factors of inflammation are examined. Sprague-Dawley rats (N = 24) underwent surgical procedures and were divided into two groups — control group and a hemorrhagic stroke group — an injection of 2 µL of sterile saline and 2 µL of 0,2U of Collagenase-IV (S) respectively. White blood cel...

  10. Anti-tumor effect of low dose total (or half) body irradiation and changes of the functional subset of peripheral blood lymphocytes in non-Hodgkin's lymphoma patients after TBI (HBI)

    Two-color analyses of peripheral blood lymphocytes using flow cytometry in 24 patients with non-Hodgkin's lymphoma, who received low dose total body irradiation (TBI) or half body irradiation (HBI), were performed to look at the influence of low dose irradiation on lymphocytes and its anti-tumor effects. The results of the present study were; significant increase in the proportion of the helper T, helper-inducer T and active helper/inducer T cells. Since low dose TBI or HBI preceded the other treatments such as primary site irradiation and multiple agent chemotherapy in 10 cases (group I), the antitumor effect of TBI or HBI were able to be investigated. Nine out of 10 patients showed at least partial responses, especially in cases 2 and 10 of group I where almost all tumors disappeared after only TBI or HBI. (author)