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Peripheral Blood (peripheral + blood)
Kinds of Peripheral Blood Terms modified by Peripheral Blood Selected AbstractsRANK Expression as a Cell Surface Marker of Human Osteoclast Precursors in Peripheral Blood, Bone Marrow, and Giant Cell Tumors of BoneJOURNAL OF BONE AND MINERAL RESEARCH, Issue 9 2006Gerald J Atkins Abstract RANK expression in vivo on hematopoietic subsets including pre-osteoclasts, identified by monoclonal antibodies, has not been described. We describe the lineages that express RANK in bone marrow, peripheral blood, and GCTs. We show that CD14+RANKhigh cells constitute a circulating pre-osteoclast pool. Introduction: The expression of RANK by subsets of hematopoietic cells has not been adequately studied in humans. While attributed to the monocytoid lineage, the phenotype of the pre-osteoclast (pre-OC) with respect to RANK expression in vivo remains unclear. We tested monoclonal antibodies (MAbs) raised against the extracellular domain of recombinant human RANK for reactivity with normal peripheral blood (PB) and bone marrow (BM) mononuclear cells (PBMNCs and BMMNCs, respectively). We also tested reactivity with giant cell tumor cells (GCT), a confirmed source of pre-OC and mature OCs. Materials and Methods: Human PBMNCs, BMMNCs, and GCT cells were analyzed for reactivity with anti-RANK MAbs by flow cytometry in combination with hematopoietic lineage restricted markers. GCTs were also analyzed by immunofluorescence. CD14+ monocytoid cells were sorted by fluorescence-activated cell sorting (FACS) based on their relative RANK expression and cultured under OC-forming conditions. Results: RANK+ cells were detected similarly by three independent anti-RANK MAbs. One MAb (80736) immunoprecipitated RANK,RANKL complexes from surface-biotinylated GCT lysates. Using dual-color flow cytometry, RANK was detected on CD14+ (monocytoid), CD19+ (B-lymphoid), CD56+ (NK cell), and glycophorin A+ erythroid progenitors. Minor populations of both CD3+ T lymphocytes and BM CD34+ hematopoietic progenitors also expressed cell surface RANK. In GCTs, RANK expression was identified on mononuclear CD45+CD14+,V,3+c-Fms+ cells, likely to be committed pre-OC, and on multinucleated CD45+,V,3+TRACP+ OCs. Importantly, sorted CD14+RANKhigh PBMNCs treated with recombinant RANKL and macrophage-colony stimulating factor (M-CSF) gave rise to approximately twice the number of osteoclasts than RANKmid or RANKlow cells. Conclusions: These results suggest that committed monocytoid RANK+ pre-OCs are represented in the marrow and circulate in the periphery, forming a pool of cells capable of responding rapidly to RANKL. The ability to reliably detect committed pre-OC in peripheral blood could have important clinical applications in the management of diseases characterized by abnormal osteoclastic activity. [source] Granulocyte colony-stimulating factor produces a decrease in IFN, and increase in IL-4 when administrated to healthy donorsJOURNAL OF CLINICAL APHERESIS, Issue 4 2010Octavio Rodríguez-Cortés Abstract Hematopoietic stem cells transplantation (HSCT) is the leading curative therapy for a variety of hematological and hereditary diseases; however, graft versus host disease (GVHD), an immunologic phenomenon that is favored by Th1 cytokines and cytotoxic cells from donors, is present frequently and is one of the most important causes of transplant related mortality. Peripheral blood HSCT is the preferred source of stem cells in almost 100% of the cases of autologous HSCT and in 70% of allogeneic transplants. The best mobilizing agent to get the stem cells out from the bone marrow is the Granulocyte-Colony Stimulating Factor (G-CSF). In this work, our main objective was to study a possible correlation between the graft cell dose and the patient's clinical outcome. We evaluated the immunologic changes produced by G-CSF in the lymphocyte and cytokine profiles in allogeneic HSC donors. HSC from twelve donors were mobilized with G-CSF at 16 ,g/kg/day, for 5 days. Basal Peripheral Blood (BPB), Mobilized Peripheral Blood (MPB), and aphaeresis mononuclear cells (G-MNC) samples were taken from all donors. Using flow cytometry, we quantified CD19+, CD3+, CD3+CD4+, CD3+CD8+, NK, NKT, DC1, and DC2 cells. Cytokines were determined by ELISA in culture supernatants. CD19+ (p = 0.001), DC1 (p < 0.002) and DC2 (p < 0.001) cells were increased in MPB with respect to BPB. An increase in Th2 cytokines such as (IL-4) and a decrease in Th1 cytokines (IFN,, IL-2) were also found in MPB samples. In conclusion, Th1 and Th2 cytokines are relevant in predicting the clinical outcome after allogeneic peripheral blood HSCT. J. Clin. Apheresis 25:181,187, 2010. © 2010 Wiley-Liss, Inc. [source] Relationships Between Concentrations of Cocaine and Its Hydrolysates in Peripheral Blood, Heart Blood, Vitreous Humor and UrineJOURNAL OF FORENSIC SCIENCES, Issue 2 2006Wayne C. Duer Ph.D. ABSTRACT: Cocaine is known to degrade in vivo and in vitro by several hydrolytic mechanisms. A previous study found that the initial amount of cocaine added to plasma could be accounted for by summing the molar concentrations of cocaine's hydrolysis products and the cocaine remaining after hydrolysis. The present study was undertaken to investigate whether or not relationships might exist between such molar concentration sums for different postmortem bodily fluids. Determinations of cocaine, benzoylecgonine, ecgonine methyl ester, and ecgonine were performed using liquid chromatography/mass spectrometry (LC/MS/MS) with heart blood, femoral blood, vitreous humor (VH), and urine (UR). The results demonstrate a strong correlation between blood and VH concentrations (correlation coefficients of 0.88,0.94), weak correlation between the UR and blood concentrations (correlation coefficients of 0.61,0.64), and weak correlation between UR and VH concentrations (correlation coefficient of 0.59). The results demonstrate that ecgonine is a significant hydrolysate with concentrations on the same order of magnitude as benzoylecgonine. The results are consistent with rapid distribution of the parent drug and its hydrolysates in the blood and VH. The strong correlation between the blood and VH demonstrates that VH is an important medium for toxicology testing when attempting to make a determination of cocaine intoxication. [source] The Application of Magnetic Cell Sorter (MACS) to Detect Fetal Cells in Maternal Peripheral BloodJOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH (ELECTRONIC), Issue 3 2001Dr. Akimune Fukushima Abstract Objectives: The aim of the present study was to investigate the effectiveness of sorting fetal nucleated red blood cells (FNRBC) from maternal peripheral blood, particularly during early gestation periods, by a combination of specific gravity centrifugation and magnetic cell sorter (MACS). Methods: Without prior knowledge of the gender of the fetus, we determined gender by analyzing a Y-chromosome specific sequence by nested-PCR, using 10 ml of the peripheral blood of healthy primigravida women at different stages of gestation (first trimester: n = 17, second trimester: n = 13, and third trimester: n = 19). The results of this prenatal sex determination were compared to the sex of newborns. Results: The specificity, sensitivity, positive predictive value (PPV) and negative predictive value (NPV) of the present method during the first trimester were 100, 81.8, 100, and 75%, respectively; during the second trimester, 80, 50, 80, and 50%, respectively; and during the third trimester, 25, 63.6, 53.8, and 33.3%, respectively. Conclusion: The results show that this prenatal sex determination method has a highly accurate diagnostic rate during the first trimester, suggesting that it could be developed as a practical, non-invasive prenatal diagnostic technique for use during early gestation periods. [source] Elevated NK Cell Cytotoxicity, CD158a Expression in NK Cells and Activated T Lymphocytes in Peripheral Blood of Women with IVF FailuresAMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 1 2010Viktor P. Chernyshov Citation Chernyshov VP, Sudoma IO, Dons'koi BV, Kostyuchyk AA, Masliy YV. Elevated NK cell cytotoxicity, CD158a expression in NK cells and activated T lymphocytes in peripheral blood of women with IVF failures. Am J Reprod Immunol 2010; 64: 58,67 Problem, The aim of this study was to evaluate the role of elevated natural killer cytotoxicity (NKc) in women with multiple implantation failures (IF) in vitro fertilization,embryo transfer (IVF,ET) cycles. Methods of study, Seventy-nine antiphospholipid antibodies-negative women with IF including 33 women with elevated NKc were selected for investigation. K-562 cell line was used to evaluate NKc. Lymphocyte subsets, intracellular cytokines [interferon (IFN)-,, interleukin (IL)-4, tumour necrosis factor, IL-10], expression of activating markers [CD69, human leukocyte antigen (HLA)-DR], CD8, KIR (CD158a), CD95, and chemokine receptors (CXCR3, CCR4) were estimated by flow cytometry. Results, In women with IF, levels of NKc were higher than in IVF successful women. IF was associated with higher expression of CD8, CD158a, and HLA-DR in NK cells, activating markers in T lymphocytes, and lower levels of CCR4+ and IL-4+ T lymphocyte subsets. Predictive value of single elevated NKc for IVF success was 0.85, but addition of two other abnormal parameters resulted in its decrease to <0.39. Conclusions, Elevated NKc is negative factor, though not critical for implantation in IVF cycles. Immune mechanism of IVF failure includes not only elevated NKc but also some other factors, such as elevated expression of CD8 and CD158a on NK cells, T lymphocyte activation, and diminished T helper 2 parameters. [source] Dendritic Cell Subset Ratio in Peripheral Blood Correlates with Successful Withdrawal of Immunosuppression in Liver Transplant PatientsAMERICAN JOURNAL OF TRANSPLANTATION, Issue 6 2003George V. Mazariegos Human dendritic cell (DC) subsets appear to play distinct roles in the induction and regulation of immune responses. While monocytoid DC (DC1) induce T-helper (Th) 1-type responses, plasmacytoid DC (DC2) have been reported to selectively induce Th2 responses. In blood, their precursors (p) can be identified as HLA-DR+ lineage, cells that are further characterized as CD11c+ CD123,/lo (IL-3R,,/lo) (pDC1) or as CD11c, CD123hi (pDC2) by rare event, flow cytometric analysis. We compared the incidences of pDC1 and pDC2 in peripheral blood mononuclear cell populations isolated from normal healthy controls and from 3 groups of clinically stable liver transplant patients. Group A had been successfully withdrawn from immunosuppression, whereas group B were undergoing prospective drug weaning and on minimal anti-rejection therapy. In group C, drug withdrawal had either failed or never been attempted and patients were on maintenance immunosuppression. Assessment of DC subsets and the pDC2 : pDC1 ratio showed good intra-and interassay reproducibility. Compared with patients in group C, those in groups A and B demonstrated a significantly higher relative incidence of pDC2 and a lower incidence of pDC1 , similar to those values observed in normal healthy controls. Moreover, the pDC2 : pDC1 ratio was significantly higher in patients undergoing (successful) weaning and in those off immunosuppression compared with patients on maintenance immunosuppression. [source] Intestinal double-positive CD4+CD8+ T,cells are highly activated memory cells with an increased capacity to produce cytokinesEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 3 2006Bapi Pahar Dr. Abstract Peripheral blood and intestinal CD4+CD8+ double-positive (DP) T,cells have been described in several species including humans, but their function and immunophenotypic characteristics are still not clearly understood. Here we demonstrate that DP T,cells are abundant in the intestinal lamina propria of normal rhesus macaques (Macaca mulatta). Moreover, DP T,cells have a memory phenotype and are capable of producing different and/or higher levels of cytokines and chemokines in response to mitogen stimulation compared to CD4+ single-positive T,cells. Intestinal DP T,cells are also highly activated and have higher expression of CCR5, which makes them preferred targets for simian immunodeficiency virus/HIV infection. Increased levels of CD69, CD25 and HLA-DR, and lower CD62L expression were found on intestinal DP T,cells populations compared to CD4+ single-positive T,cells. Collectively, these findings demonstrate that intestinal and peripheral blood DP T,cells are effector cells and may be important in regulating immune responses, which distinguishes them from the immature DP cells found in the thymus. Finally, these intestinal DP T,cells may be important target cells for HIV infection and replication due to their activation, memory phenotype and high expression of CCR5. [source] Analysis of chimerism during the early period after allogeneic peripheral stem cell transplantationINTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY, Issue 6 2001B. Gleissner As there are few reports on early evaluation of chimerism, we assessed fluorescence short tandem repeats (STR) by polymerase chain reaction (PCR) assays to analyse donor and recipient characteristics at early time points after peripheral stem cell transplantation (PBSCT). Peripheral blood of 13 patients was analysed in 1- to 2-day intervals starting from the day of PBSCT. Donor and recipient allelic patterns were determined by a commercially available multiplex STR assay that simultaneously evaluates four or five gene loci. Mixed chimerism appeared in all patients during days 1,9 after transplantation and preceded haematologic engraftment for 3,12 days. Even patients without myeloablative conditioning therapy (n=4) revealed donor allelic patterns within 1,5 days. Nine patients changed during the following days to a complete donor allelic pattern and had an uncomplicated post-transplant disease course. Four patients did not consistently retain complete donor chimerism; two of them relapsed within the next 3 months, one died from septicemia within 7 days, and the fourth, transplanted for aplastic anaemia, is still in complete remission. Overall, STR analysis using a simple and comparatively cheap multiplex system permits the detection of chimerism very early after transplantation and may provide relevant information that correlates with the clinical follow-up. [source] The Effects of Ethanol Consumption on Vasculogenesis Potential in Nonhuman PrimatesALCOHOLISM, Issue 1 2008J. Koudy Williams Background:, Vasculogenesis is essential to the preservation and repair of damaged or diseased vessels. Alcohol is the most commonly abused drug among young adults, but its effects on vessel growth and repair are unknown. The basis of vascular repair is endothelial progenitor cell (EPC) recruitment to assist in the formation of new vascular network (vasculogenesis). Therefore, the objective of this study was to measure the effects of ethanol consumption on the production, mobilization and vasculogenesis potential EPCs in nonhuman primates. Methods:, Four to five year-old (young adult) male rhesus monkeys consumed monkey chow and water (Control, n = 7), or chow and water + ethanol (Alcohol, 2.45 g/d, n = 7) for 12 months. Peripheral blood (PB) and bone marrow (BM) samples were collected for fluorescence-activated cell-sorting analysis of cell surface antigens (CD45, CD31, CD44, CD133, VEGF-R2 , or KDR); and for capillary formation on Matrigel-coated plates. Results:, There were greater numbers of nonhematopoeitic stromal cells (CD45,) and putative mesenchymal progenitor cells (CD45,/CD44+) in the PB and BM of Alcohol versus Control monkeys (p < 0.05). Additionally, there were greater numbers of EPCs (CD45,/CD133+/KDR+) in the BM and PB of Alcohol versus Control monkeys (p < 0.05). However, the EPCs of Alcohol monkeys were less likely to form capillaries on matrigel-coated plates than Control monkeys (p < 0.05). Conclusions:, Ethanol consumption in monkeys markedly increased the production and mobilization of EPCs, but decreased their ability to form capillaries. The pathophysiologic consequences of such effects are unclear, but may represent an ethanol-induced chronic stress on the BM, resulting in EPC. [source] Surgery-related shedding of breast cancer cells as determined by RT-PCR assayJOURNAL OF SURGICAL ONCOLOGY, Issue 4 2003Xi-Chun Hu MD Abstract Background and Objectives Surgery could result in the shedding of cancer cells into the circulation. These cells were investigated with reverse transcriptase-polymerase chain reaction (RT-PCR) assay for cytokeratin 19 (CK19) and ,-subunit of human chorionic gonadotropin (,-hCG). Patients and Methods Peripheral blood was sampled from 49 patients with breast cancer before operation (d,1), 1 day after operation (d1), and 7 days after operation (d7). Total RNA was extracted from peripheral blood mononuclear cells, followed by RT-PCR assay. The products for ,-hCG were digested with Sty I endonuclease. The patients were followed up for a median of 33 months for signs of recurrence and metastasis. Results The results for CK19 at d,1, d1, and d7 were 8.2, 20.4, and 10.2%, respectively. For ,-hCG, the corresponding results were 12.2, 26.5, and 16.3%, respectively. There was a higher positive rate in d1 samples than in d,1 samples for CK19 and ,-hCG (P,<,0.05 and P,=,0.092, respectively). Conversions of signals from being negative to positive were found in all stages. These did not demonstrate a statistical correlation with prognostic factors associated with a poor prognosis. Only two of the five recurrence occurred in the 15 patients with the signal conversions, while the other three occurred in the patients showing no signals in all samples. Conclusions Cancerous breast cells that enter into the blood circulation as a result of an operation are unlikely to be involved in the formation of metastatic deposits. J. Surg. Oncol. 2003;82:228,232. © 2003 Wiley-Liss, Inc. [source] Rapid method for the analysis of peripheral chimerism in suspected graft-versus-host disease after liver transplantationLIVER TRANSPLANTATION, Issue 2 2000Amy B. Hahn The effects of microchimerism and possible tolerance have been well studied in orthotopic liver transplantation. In some patients, greater levels of donor cells persist in the periphery. These cells were characterized and their effects on clinical outcome were studied. Peripheral blood was obtained from patients at various times posttransplantation. HLA class II typing was performed by the polymerase chain reaction,sequence-specific primer method on unfractionated blood and lymphocyte subpopulations. Relative levels of amplification of donor and recipient alleles were compared. All patients studied had a low degree of chimerism that was most apparent in the CD8+T/natural killer (NK) cell population. One patient with persistently high levels of donor alleles in his CD8+T/NK cell population was diagnosed with severe graft-versus-host disease (GVHD) and died of opportunistic infections. Another patient with biopsy-proven GVHD was chimeric in several cell populations. On resolution of her symptoms, donor alleles were reduced to levels undetectable by this assay. These results suggest that persistently elevated levels of donor CD8+T/NK cells in the periphery may indicate GVHD in liver transplant recipients. This technique aids in rapid diagnosis, which facilitates appropriate treatment and thus may improve clinical outcome. [source] Peripheral blood and bone marrow morphology in adult T-cell leukaemia/lymphoma,AMERICAN JOURNAL OF HEMATOLOGY, Issue 7 2010Kikkeri N. Naresh No abstract is available for this article. [source] Transcriptosome and serum cytokine profiling of an atypical case of myelodysplastic syndrome with progression to acute myelogenous leukemiaAMERICAN JOURNAL OF HEMATOLOGY, Issue 10 2006Daruka Mahadevan Abstract A Native American-Indian female presenting with anemia and thrombocytosis was diagnosed with myelodysplastic syndrome (MDS, refractory anemia). Over the course of 5 years she developed cytopenias and periods of leukocytosis with normal bone marrow (BM) blast counts, features of an unclassifiable MDS/MPS syndrome. The patient ultimately progressed to acute myelogenous leukemia (AML, FAB M2) and had a normal karyotype throughout her course. The episodes of leukocytosis were associated with infectious complications. Transformation to AML was characterized by a BM blast percentage of 49%. Peripheral blood and BM samples were obtained for serum protein analysis and gene expression profiling (GEP) to elucidate her disease process. An ELISA assay of the serum analyzed ,80 cytokines, which demonstrated that hepatocyte growth factor/scatter factor and insulin-like growth factor binding protein 1 were markedly elevated compared to normal. GEP demonstrated a unique "tumor molecular profile," which included overexpression of oncogenes (HOXA9, N-MYC, KOC1), proliferative genes (PAWR, DLG5, AKR1C3), invasion/metastatic genes (FN1, N-CAM-1, ITGB5), pro-angiogenesis genes (c-Kit), and down regulation of tumor suppressor genes (SUI1, BARD1) and anti-apoptotic genes (PGLYRP, SERPINB2, MPO). Hence, a biomics approach has provided insight into elucidating disease mechanisms, molecular prognostic factors, and discovery of novel targets for therapeutic intervention. Am. J. Hematol., 2006. © 2006 Wiley-Liss, Inc. [source] Effect of a combination of extract from several plants on Cell-mediated and humoral immunity of patients with advanced ovarian cancerPHYTOTHERAPY RESEARCH, Issue 5 2006N. Kormosh Abstract The influence of a plant preparation AdMax (Nulab Inc., Clearwater, FL, USA) on immunity in ovarian cancer patients was studied. The preparation is a combination of dried ethanol/water extracts from roots of Leuzea carthamoides, Rhodiola rosea, Eleutherococcus senticosus and fruits of Schizandra chinensis. Twenty eight patients with stage III,IV epithelial ovarian cancer were treated once with 75 mg/m2 cisplatin and 600 mg/m2 cyclophosphamide. Peripheral blood was collected 4 weeks after the chemotherapy. Subclasses of T, B and NK lymphocytes were tested for in the blood samples: CD3, CD4, CD5, CD7, CD8, CD11B, CD16, CD20, CD25, CD38, CD45RA, CD50, CD71 and CD95. Immunoglobulin G, A and M concentrations were also determined. Changes were observed in the following T cell subclasses: CD3, CD4, CD5 and CD8. In patients who took AdMax (270 mg a day) for 4 weeks following the chemotherapy, the mean numbers of the four T cell subclasses were increased in comparison with the mean numbers of the T cell subclasses in patients who did not take AdMax. In patients who took AdMax, the mean amounts of IgG and IgM were also increased. The obtained results suggest that the combination of extracts from adaptogenic plants may boost the suppressed immunity in ovarian cancer patients who are subject to chemotherapy. Copyright © 2006 John Wiley & Sons, Ltd. [source] Applying a test system for discriminating fetal from maternal cellsPRENATAL DIAGNOSIS, Issue 8 2003nar Bayrak-Toydemir Abstract Objective The objectives of this study were to enhance and apply a simple system capable of testing the capacity of putative, gender-independent fetal cell markers, individually and in combination, to discriminate between fetal and maternal cells. Methods Chorionic villi tissue obtained from 25 male pregnancies at 10 to 12 weeks' gestation served as the experimental group. Following removal of villi pieces for clinical use, unattached cells were collected by centrifugation of the CVS fluid, fixed in the tube, and used as a source of mixed fetal and maternal cells. Blood obtained from a fetus at 13 weeks' gestation served as a positive control. Peripheral blood from two adult males served as negative controls. Antibodies to three possible fetal markers were tested using immunohistochemical techniques: anti-Flk-1, anti-epsilon globin, and anti-CD71. Each antibody was used alone and in combination in conjunction with fluorescent in situ hybridization (FISH) of X and Y chromosomes to confirm that positively stained cells were in fact fetal in origin. Results On CVS samples, the average predictive value for anti-Flk-1 was 35.8%, 76.2% for anti-CD71, and 90.5% for anti-epsilon. The combination of anti-epsilon and anti-CD71 antibodies identifying a fetal cell was 87.2% and the combined use of single and double antibodies gave a value of 82.7%. The combination of anti-epsilon globin and anti-CD71 increased the sensitivity of identifying pure fetal blood cells from 63%, for anti-epsilon alone, and 67%, for anti-CD71 alone, to 86%. Conclusion Although anti-Flk-1 has been reported to be a successful marker of fetal cells, the results in this test system did not support this finding. This work supports the use of CVS washings containing both fetal and maternal cells as a viable test system for assessing antigenic markers. The combination of anti-CD71 and anti-epsilon as fetal identifiers may increase the chances of identifying a fetal cell without compromising the predictive value. Copyright © 2003 John Wiley & Sons, Ltd. [source] 1141154113 Expression of natural cytotoxicity receptors in peripheral blood NK cell subsets of women with recurrent spontaneous abortions (RSA) or implantation failuresAMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 6 2006A Fukui Problem:, Natural Cytotoxicity Receptors (NCRs) are unique markers of NK cells and regulate NK cell cytotoxicity and cytokine production. a2V-ATPase is expressed in the cell membrane and can regulate the pH of the extracellular environment, which might facilitate NK cell killing or cytokine secretion. In this preliminary study we evaluated the expression of NCRs and a2V-ATPase in peripheral blood NK cells of women with RSA or implantation (IVF-ET) failures. Method of Study:, Peripheral blood was obtained from women with RSA (n = 10), or IVF-ET failures (n = 9). CD56dim and CD56bright NK cells were analyzed for the expression of NCRs (NKp46, NKp44 and NKp30) and a2V-ATPase using flow cytometry. Results:, For women with RSA, there were significant differences in the expression of NKp46 between CD56dim (36.9 ± 30.2) and CD56bright (76.0 ± 27.5) (P < 0.01), of NKp30 between CD56dim (30.9 ± 25.7) and CD56bright (55.8 ± 29.5) (P < 0.01), and of a2V-ATPase between CD56dim (1.0 ± 0.9) and CD56bright (23.2 ± 15.1) (P < 0.01) NK cells. For women with IVF-ET failures, there were significant differences in the expression of NKp46 between CD56dim (39.5 ± 21.5) and CD56bright (78.8 ± 26.0) (P < 0.01), of NKp30 between CD56dim (27.2 ± 17.9) and CD56bright (45.2 ± 29.8) (P < 0.05), and of a2V-ATPase between CD56dim (1.6 ± 1.4) and CD56bright (21.2 ± 16.5) (P < 0.01) NK cells. Conclusions:, The differential expression of NCRs and a2V-ATPase in NK cell subsets of women with RSA and IVF-ET failures may have an effect in cytotoxicity and cytokine production. Additional studies are currently in effect to evaluate these activities. We suggest that the analysis of NCRs and a2V-ATPase expression in peripheral blood NK cell subsets may contribute to a better understanding in the biology of NK cells in women with RSA or IVF-ET failures. [source] Maternal and Neonatal Lymphocyte Subpopulations at Delivery and 3 Days Postpartum: Increased Coexpression of CD45 IsoformsAMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 1 2004Emilia Juretic Problem:, Huge physiologic changes during parturition involve immune cells. Alterations in maternal and neonatal lymphocytes postpartum might ascertain the subpopulations that are most affected and may possibly be of importance in the process. Method of study:, Peripheral blood was taken from 20 healthy women at vaginal delivery and 3 days later, concomitantly with cord and peripheral blood from their newborns. Lymphocyte immunophenotyping was done by three-color flow-cytometry. Results:, Maternal T helper cells were decreased and natural killer (NK) cells were significantly increased during labor. Percentage of CD4+ and percentage and absolute count of CD8+ cells coexpressing CD45RA and CD45RO antigens were higher than 3 days later. In cord blood NK cells were considerably increased and more CD4+ cells expressed CD45RO antigen. Conclusion:, Coexpression of CD45RA and CD45RO molecules indicates activation of maternal CD4+ and CD8+ lymphocytes. NK cells increase suggests their possible association with parturition processes. Lymphocyte subsets in cord blood correspond to maternal subsets to some extent. [source] Role of CD8+ CD25+ Foxp3+ regulatory T cells in multiple sclerosisANNALS OF NEUROLOGY, Issue 5 2010Jorge Correale MD Objective The objective of this study was to investigate the role of CD8+ CD25+ FoxP3+ cells during the course of multiple sclerosis (MS). Methods Peripheral blood and cerebrospinal fluid (CSF) CD8+ T-cell clones (TCCs) recognizing autoreactive CD4+ T cells were isolated from 20 MS patients during exacerbations, 15 patients in remission, 15 healthy subjects, and 10 patients with other inflammatory neurological diseases. Characteristics of noncytotoxic CD8+ CD25+ regulatory T cells were studied. Cell phenotype was evaluated using flow cytometry. Cytokine production and phospho-signal transducer and activator of transcription 3 (STAT3) concentration were determined using enzyme-linked immunosorbent assay. To assess 2,3-dioxygenase (IDO) activity on dendritic cells (DCs), kynurenine concentration was measured by high-performance liquid chromatography. Results Inhibition of CD4+ self-reactive T-cell proliferation, and of interferon-, and interleukin (IL)-17 secretion, was observed after adding CD8+ CD25+ FoxP3+ cells to cultures. Suppression was abrogated by silencing FoxP3 using small interfering RNA. Cells were CD122+, CTLA-4+, GITR+, CCR7+, and CD62L+, producing IL-10 and transforming growth factor-,. CD8+ CD25+ FoxP3+ cells downregulated costimulatory molecule expression on dendritic cells through a STAT3-mediated pathway, resulting in less efficient antigen presentation, and induced IDO expression on DCs through STAT3 and cytotoxic T-lymphocyte antigen 4-dependent mechanisms. CD8+ regulatory TCC cloning frequency studied in blood and CSF was suppressed to a greater degree during exacerbations than during remission or in controls. Likewise, in CSF of MS patients during acute exacerbations, lower levels of CD8+ CD25+ FoxP3+ T cells were detected using flow cytometry. Interpretation CD8+ CD25+ FoxP3+ cells are novel regulatory cells exerting significant influence over self-reactive CD4+ T-cell regulation during the course of MS. Induction of these cells may provide new therapeutic alternatives for MS by eliminating or inhibiting self-reactive T cells. ANN NEUROL 2010;67:625,638 [source] Increased numbers of circulating polyfunctional Th17 memory cells in patients with seronegative spondylarthritidesARTHRITIS & RHEUMATISM, Issue 8 2008Camilla Jandus Objective A distinct subset of proinflammatory CD4+ T cells that produce interleukin-17 was recently identified. These cells are implicated in different autoimmune disease models, such as experimental autoimmune encephalomyelitis and collagen-induced arthritis, but their involvement in human autoimmune disease has not yet been clearly established. The purpose of this study was to assess the frequency and functional properties of Th17 cells in healthy donors and in patients with different autoimmune diseases. Methods Peripheral blood was obtained from 10 psoriatic arthritis (PsA), 10 ankylosing spondylitis (AS), 10 rheumatoid arthritis (RA), and 5 vitiligo patients, as well as from 25 healthy donors. Synovial tissue samples from a separate group of patients were also evaluated (obtained as paraffin-embedded sections). Peripheral blood cells were analyzed by multiparameter flow cytometry and immunohistochemistry. Cytokine production was examined by enzyme-linked immunosorbent assay and intracellular cytokine staining using specific monoclonal antibodies. Synovial tissue was examined for infiltrating T cells by immunohistochemical analysis. Results We found increased numbers of circulating Th17 cells in the peripheral blood of patients with seronegative spondylarthritides (PsA and AS), but not in patients with RA or vitiligo. In addition, Th17 cells from the spondylarthritis patients showed advanced differentiation and were polyfunctional in terms of T cell receptor,driven cytokine production. Conclusion These observations suggest a role of Th17 cells in the pathogenesis of certain human autoimmune disorders, in particular the seronegative spondylarthritides. [source] Elevated CD16 expression by monocytes from patients with tumor necrosis factor receptor,associated periodic syndromeARTHRITIS & RHEUMATISM, Issue 12 2007Ian Todd Objective Tumor necrosis factor receptor,associated periodic syndrome (TRAPS) is an inherited autosomal-dominant autoinflammatory condition caused by mutations in the ectodomain of the 55-kd tumor necrosis factor (TNF) receptor superfamily 1A. Proinflammatory blood monocytes with the phenotype CD14+,CD16+,HLA,DR++ are a major source of TNF, and the number of such monocytes is increased during infection and inflammation. The aim of this study was to investigate whether the expression of circulating CD16+ monocytes is affected in patients with TRAPS. Methods Peripheral blood obtained from patients with TRAPS and healthy control subjects was stained with monoclonal antibodies to detect CD14++,CD16, monocytes and CD14+,CD16+ monocytes, using flow cytometry. Lipopolysaccharide-induced TNF production was measured by intracellular cytokine staining. Activation-induced shedding of CD16 was investigated by treating blood samples with phorbol myristate acetate. Results The level of CD16 expression by CD14+,CD16+ monocytes, but not their absolute number, was significantly elevated in patients with TRAPS, even though the patients were not experiencing clinically overt episodes of autoinflammation at the time of sampling. These findings are similar to those for the C-reactive protein levels and erythrocyte sedimentation rates in the same patients. The enhanced level of CD16 expression by monocytes from patients with TRAPS was not attributable to a defect in activation-induced shedding of CD16. The CD14+,CD16+ monocytes were the predominant source of TNF in both patients and healthy control subjects. Conclusion The level of CD16 expression by monocytes was elevated in patients with TRAPS, as a feature of the underlying constitutive inflammation status. [source] Evaluation of genotoxic effects in human leukocytes after in vitro exposure to 1950 MHz UMTS radiofrequency fieldBIOELECTROMAGNETICS, Issue 3 2008O. Zeni Abstract In the present study the third generation wireless technology of the Universal Mobile Telecommunication System (UMTS) signal was investigated for the induction of genotoxic effects in human leukocytes. Peripheral blood from six healthy donors was used and, for each donor, intermittent exposures (6 min RF on, 2 h RF off) at the frequency of 1950 MHz were conducted at a specific absorption rate of 2.2 W/kg. The exposures were performed in a transverse electro magnetic (TEM) cell hosted in an incubator under strictly controlled conditions of temperature and dosimetry. Following long duration intermittent RF exposures (from 24 to 68 h) in different stages of the cell cycle, micronucleus formation was evaluated by applying the cytokinesis block micronucleus assay, which also provides information on cell division kinetics. Primary DNA damage (strand breaks/alkali labile sites) was also investigated following 24 h of intermittent RF exposures, by applying the alkaline single cell gel electrophoresis (SCG)/comet assay. Positive controls were included by treating cell cultures with Mitomycin-C and methylmethanesulfonate for micronucleus and comet assays, respectively. The results obtained indicate that intermittent exposures of human lymphocytes in different stages of cell cycle do not induce either an increase in micronucleated cells, or change in cell cycle kinetics; moreover, 24 h intermittent exposures also fail to affect DNA structure of human leukocytes soon after the exposures, likely indicating that repairable DNA damage was not induced. Bioelectromagnetics 29:177,184, 2008. © 2007 Wiley-Liss, Inc. [source] Regulatory T cells in atopic dermatitis: epidermal dendritic cell clusters may contribute to their local expansionBRITISH JOURNAL OF DERMATOLOGY, Issue 5 2009A. Szegedi Summary Background, Regulatory T cells (Tregs) have an essential role in tolerance and immune regulation. However, few and controversial data have been published to date on the role and number of these cells in atopic dermatitis (AD). Objectives, To investigate the number of CD4+CD25+FOXP3+ Tregs and interleukin 10-producing T regulatory type 1 (Tr1) cells in patients with AD. Methods, Peripheral blood and skin biopsy samples from atopy patch test (APT)-positive patients with acute- and chronic-phase AD were investigated. Immunohistochemistry was applied to identify CD4+CD25+FOXP3+ Tregs in the skin, while flow cytometry was used to detect CD4+CD25highFOXP3+ Tregs and Tr1 cells in the peripheral blood. Results, In the peripheral blood samples of patients with AD significantly elevated numbers of Tr1 cells were found. Although neither the absolute number nor the percentage of CD4+CD25highFOXP3+ Tregs showed significant alteration in the peripheral blood of patients, increased numbers of FOXP3+ Tregs were detected in skin biopsy specimens. All of the APT-positive skin samples showed epidermal dendritic cell aggregates, morphologically consistent with so-called Langerhans cell microgranulomas, which also contained intermingled FOXP3+ Tregs. Conclusions, Tr1 cell numbers were elevated in the peripheral blood and increased numbers of CD4+CD25highFOXP3+ Tregs were detected in the skin of patients with AD. The epidermal dendritic cell clusters in APT-positive lesional skin showed a close connection to the FOXP3+ Tregs. [source] The presence of dominant T-cell clones in peripheral blood of patients with collagen vascular disorders: a prospective study of 97 casesBRITISH JOURNAL OF DERMATOLOGY, Issue 3 2006O. Dereure Summary Background, T-lymphocyte dysfunction has been seldom investigated in collagen vascular disorders. The search for dominant T-cell clones has been scarcely reported, although the presence of such clones might be expected in disorders showing immune responses directed against a variety of autoantigens. Objectives, We conducted a systematic search for dominant T-cell clones in peripheral blood in patients with collagen vascular disorders. Patients and methods, Ninety-seven patients with collagen vascular disorders were studied (7 cutaneous and 38 systemic lupus erythematosus; 8 multiple morphea; 12 regional scleroderma; 32 systemic sclerosis of the CREST type). A dominant T-cell clone was searched for in peripheral blood by polymerase chain reaction targeting the T-cell receptor , chain followed by a size analysis of amplified fragments. Peripheral blood from patients with nonlymphocyte-dependent disorders and matched by age and sex was assessed in the same conditions. Results in both groups were compared using nonparametric statistical tests. Results, Overall, a circulating dominant T-cell clone was found in 52% of patients compared with 16·9% in controls. More precisely, such a dominant clone was present in 43% and 37% of cutaneous and systemic lupus erythematosus, respectively, in 75% of multiple morphea, 75% of regional scleroderma and 60% of CREST syndrome patients. The percentages in all subsets of patients were significantly higher than in the control group. Conclusions, The presence of a dominant T-cell clone in peripheral blood is significantly more frequent in collagen vascular disorders than in controls, especially in patients with scleroderma, whatever the clinical subset, which suggests T-cell involvement in the immune response dysfunction in these diseases classically characterized by disturbances of B lymphocytes. The relevance of such a dominant clone regarding diagnosis, pathomechanisms, long-term outcome and visceral prognosis of these diseases as well as therapeutic decisions remains to be evaluated. [source] Psychosocial factors and interleukin-6 among women with advanced ovarian cancerCANCER, Issue 2 2005Erin S. Costanzo M.A. Abstract BACKGROUND Relations among psychological stress, depression, social support, and interleukin-6 (IL-6, a proinflammatory cytokine) have been documented in humans and animals. Because elevated IL-6 is associated with a poorer prognosis among ovarian cancer patients and has been implicated in the metastasis of ovarian cancer, the current study examined relations between psychosocial factors and IL-6 among women with advanced-stage ovarian cancer. METHODS Sixty-one ovarian cancer patients completed assessments of social support, distressed mood, and quality of life before surgery. Peripheral blood was drawn preoperatively, and the plasma was assayed for IL-6. Ascites samples were also assayed for IL-6 for a subset of patients. RESULTS Both IL-6 levels and distressed mood were elevated among patients. After statistically adjusting effects of age and disease stage, social attachment was associated with lower levels of IL-6 in peripheral blood (P = 0.03), whereas poorer health-related quality of life was associated with higher IL-6 (P values ranged from 0.01 to 0.03 on different measures). This pattern of relations was also found in the ascites. Moreover, IL-6 levels in peripheral blood plasma correlated significantly with IL-6 in the ascites (P < 0.001), suggesting that peripheral IL-6 reflects IL-6 levels at the site of the tumor. CONCLUSIONS Results suggest that social support may play a protective role with respect to IL-6 elevations, and IL-6 may be an independent marker of health-related quality of life among ovarian cancer patients. Processes involving IL-6 represent possible pathways by which behavioral factors may contribute to disease outcomes among women with ovarian cancer. Cancer 2005. © 2005 American Cancer Society. [source] Labour increases the surface expression of two toll-like receptors in the cord blood monocytes of healthy term newbornsACTA PAEDIATRICA, Issue 6 2009Chung-Min Shen Abstract Aim: Mode of delivery may influence the innate immune system in newborns. We investigated the effect of maternal labour on the expression of two toll-like receptors, TLR2 and TLR4, in monocytes obtained from healthy full-term newborns. Methods: Monocytes were obtained from cord blood of 48 newborns that have been vaginally delivered (VD) and 14 newborns delivered by elective caesarean section (CS) without labour. Peripheral blood was also obtained from 17 healthy adults. Surface expression of TLR2 and TLR4 in the monocytes was measured by antihuman TLR2 or TLR4 monoclonal antibody and immunofluorescence flow cytometry. TLR2 and TLR4 mRNA levels were evaluated by real-time PCR. Results: CS newborns had a significantly lower level of TLR2 and TLR4 surface expression on monocytes than VD newborns. No significant difference was found between the surface expression of VD newborns and healthy adults. TLR2 and TLR4 mRNA levels in monocytes did not vary among the three study groups. Conclusion: Labour may up-regulate TLR2 and TLR4 on the cord blood monocytes of newborns at the protein level. Since TLRs are an important part of the innate immune system, our findings suggest that labour may be immunologically beneficial to normal newborns. [source] Evidence of in vivo basophil activation in chronic idiopathic urticariaCLINICAL & EXPERIMENTAL ALLERGY, Issue 6 2006K. Vasagar Summary Background Approximately 40% of chronic idiopathic urticaria (CIU) subjects have autoantibodies to either Fc,RI, or IgE. The effect of such autoantibodies on circulating basophil activation status is unknown. Objective The expression of cell surface activation markers on basophils from CIU, non-allergic, and allergic subjects were compared. Further, the relationship between marker expression and serum factors reported in CIU, such as histamine-releasing activity (HRA) and immunoreactivity to Fc,RI, were examined. Methods Peripheral blood was obtained from CIU, allergic, and non-allergic donors and fractionated by density gradients. Enriched basophils (1,12%) were analysed by flow cytometry for expression of activation markers including CD63, CD69, and CD203c. Dilutions of serum (5,50%) were analysed for HRA on basophils from a normal donor. Serum was tested for immunoreactivity by western blotting to a standard cell lysate prepared from an RBL-SX38 cell line transfected with human Fc,RI,. Results CIU subjects (n=9) and allergic subjects (n=8) exhibited enhanced expression of CD63 and CD69, as compared with non-allergic subjects (n=7); however, no difference was seen among groups for CD203c expression. Five CIU and two non-allergic subjects had evidence of significant serum HRA (>20%), whereas two CIU, two allergic, and three non-allergic subjects had evidence of serum immunoreactivity to Fc,RI,. Serum HRA and serum immunoreactivity to Fc,RI, were not associated with enhanced surface marker expression. Conclusion Basophil activation marker expression is increased in CIU subjects and is not associated with serum factors. In addition, serum HRA and Fc,RI, immunoreactivity are not unique to CIU, or related to enhanced circulating basophil marker expression. [source] Combined effect of IL-17 and blockade of nitric oxide biosynthesis on haematopoiesis in miceACTA PHYSIOLOGICA, Issue 1 2010A. Krsti Abstract Aim:, The study was undertaken to extend our investigation concerning both the in vivo activity of interleukin (IL)-17 and the specific role of nitric oxide (NO) in IL-17-induced effects in the process of haematopoiesis. Methods:, CBA mice were simultaneously treated with IL-17 and/or nitric oxide synthase (NOS) inhibitor, l -NAME, for 5 days and changes within various haematopoietic cell lineages in bone marrow, spleen and peripheral blood were analysed. Results:, Findings showed that administration of both IL-17 and l -NAME stimulated increase in net haematopoiesis in normal mice. IL-17-enhanced myelopoiesis was characterized by stimulation of both femoral and splenic haematopoietic progenitor cells and morphologically recognizable granulocytes. Additionally, IL-17 induced alterations in the frequency of erythroid progenitor cells in both bone marrow and spleen, accompanied with their mobilization to the peripheral blood. As a consequence of these changes in the erythroid cell compartments, significant reticulocytosis was observed, which evidenced that in IL-17-treated mice effective erythropoiesis occurred. Exposure of mice to NOS inhibitor also increased the number of both granulocyte-macrophage and erythroid progenitors in bone marrow and spleens, and these alterations were followed by the mobilization of erythroid progenitors and elevated content of reticulocytes in peripheral blood. The specific role of NO in IL-17-induced haematopoiesis was demonstrated only in the IL-17-reducing effect on bone marrow late stage erythroid progenitors, CFU-E. Conclusion:, The results demonstrated the involvement of both IL-17 and NO in the regulation of haematopoietic cell activity in various haematopoietic compartments. They further suggest that IL-17 effects are differentially mediated depending on the haematopoietic microenvironments. [source] Human peripheral blood B-cell compartments: A crossroad in B-cell traffic,CYTOMETRY, Issue S1 2010M. Perez-Andres Abstract A relatively high number of different subsets of B-cells are generated through the differentiation of early B-cell precursors into mature B-lymphocytes in the bone marrow (BM) and antigen-triggered maturation of germinal center B-cells into memory B-lymphocytes and plasmablasts in lymphoid tissues. These B-cell subpopulations, which are produced in the BM and lymphoid tissues, recirculate through peripheral blood (PB), into different tissues including mucosa and the BM, where long-living plasma cells produce antibodies. These circulating PB B-cells can be classified according to their maturation stage into i) immature/transitional, ii) naïve, and iii) memory B-lymphocytes, and iv) plasmablasts/plasma cells. Additionally, unique subsets of memory B-lymphocytes and plasmablasts/plasma cells can be identified based on their differential expression of unique Ig-heavy chain isotypes (e.g.: IgM, IgD, IgG, IgA). In the present paper, we review recent data reported in the literature about the distribution, immunophenotypic and functional characteristics of these cell subpopulations, as well as their distribution in PB according to age and seasonal changes. Additional information is also provided in this regard based on the study of a population-based cohort of 600 healthy adults aged from 20 to 80 years, recruited in the Salamanca area in western Spain. Detailed knowledge of the distribution and traffic of B-cell subsets through PB mirrors the immune status of an individual subject and it may also contribute to a better understanding of B-cell disorders related to B-cell biology and homeostasis, such as monoclonal B-cell lymphocytosis (MBL). © 2010 International Clinical Cytometry Society [source] CD146+ T lymphocytes are increased in both the peripheral circulation and in the synovial effusions of patients with various musculoskeletal diseases and display pro-inflammatory gene profiles,,CYTOMETRY, Issue 2 2010Pradeep Kumar Dagur Abstract Twenty-eight synovial effusions (SE) were obtained from 24 patients, paired samples of peripheral blood (PB) from 10 of these patients, and PB from 36 healthy individuals for analysis of CD146 on T-lymphocytes by flow cytometry. CD146+ or CD146, T-lymphocytes were sorted from three SE to study gene expression profiles and selected genes revalidated using QPCR assays. We found more CD3+CD146+ and CD4+CD146+ T-lymphocytes in PB from patients compared with PB of healthy individuals (4.71% ± 2.48% vs. 2.53% ± 1.08%, P = 0.028) and (6.29% ± 2.74% vs. 2.41% ± 0.96%, P = 0.0017), respectively, whereas CD8+CD146+ T-lymphocytes were not significantly different (2.55% ± 1.65% vs. 3.18% ± 2.59%, P = 0.5008). SE displayed CD146 staining on 16.32% ± 6.06% of CD3+ cells. This expression was skewed toward CD4+ T-lymphocytes, with CD146 present on 24.06% ± 8.20% of the CD4+ T-lymphocytes compared with 6.19% ± 5.22% of the CD8+ T-lymphocytes. CD146 on CD3+, CD4+ and CD8+ T-lymphocytes in SE was significantly higher compared with PB in patients (P < 0.0001, P < 0.0001 and P = 0.0036, respectively). Gene expression profiles of sorted CD146+CD4+CD3+ vs. CD146,CD4+CD3+ T-lymphocytes (n = 2) and CD2+CD146+ vs. CD2+CD 146, (n = 1) from SE, displayed increased CD146, LAIR2, CXCL13, CD109, IL6ST, IL6R, TNFRsf18, and TNFRsf4 genes, whereas decreased CCR7, CCL5, and cytotoxicity-associated genes including granzymes b, h, and k, perforin were found with the CD146, T-lymphocytes. By QPCR higher mRNA expression of CXCL13, CD146 and CD109 was also noted in the CD146+ subset, compared with the CD146, subset, in PB of healthy individuals and in PB and SE from patients. Our study establishes increased CD146+ T-lymphocytes in diseases with joint effusions, and demonstrates pro-inflammatory gene profiles in these cells. Published 2009 Wiley-Liss, Inc. [source] Harmonization of light scatter and fluorescence flow cytometry profiles obtained after staining peripheral blood leucocytes for cell surface-only versus intracellular antigens with the Fix & PermÔ reagent,,§CYTOMETRY, Issue 1 2010Elaine Sobral da Costa Abstract Staining for intracellular markers with the Fix & PermÔ reagent is associated with variations in the scatter properties of leucocytes, limiting automated analysis of flow cytometry (FCM) data. Here, we investigated those variables significantly contributing to changes in the light scatter, autofluorescence, and bcl2 staining characteristics of peripheral blood (PB) leucocytes, after fixation with Fix & PermTM. Our major aim was to evaluate a new mathematical approach for automated harmonization of FCM data from datafiles corresponding to aliquots of a sample treated with cell-surface-only versus Fix & Perm intracellular staining techniques. Overall, neither the anticoagulant used nor sample storage for <24 h showed significant impact on the light scatter and fluorescence properties of PB leucocytes; similarly, the duration of the fixation period (once >15 min were used) had a minimum impact on the FCM properties of PB leucocytes. Conversely, changes in cell/protein concentrations and the fixative/sample (vol/vol) ratio had a clear impact on the light scatter features of some populations of leucocytes. Accordingly, lower cell/protein concentrations were associated with lower scatter values, particularly for the neutrophils. Such changes could be partially corrected through the use of higher fixative to sample volume ratios. Despite the variable changes detected between aliquots of the same sample treated with cell surface-only versus intracellular staining procedures, the new mathematical approach here proposed and evaluated for automated harmonization of common parameters in both datafiles, could correct the FCM profiles of leucocytes derived from cells undergoing conventional fixation/permeabilization procedures, and made them indistinguishable from those corresponding to aliquots of the same sample treated with cell-surface-only staining techniques. © 2009 Clinical Cytometry Society [source] | ||||||||||||||||||||||||||||||||||||||||||||||