CD4-Positive T-Lymphocytes ; Child ; Cryptococcosis ; immunology ; Humans ; Lymphopenia ; etiology

Apoptosis ; Proteins/metabolism* ; Mitochondria/metabolism* ; CD4-Positive T-Lymphocytes

CD4-Positive T-Lymphocytes ; Cell Differentiation ; T-Lymphocyte Subsets ; T-Lymphocytes, Regulatory

BACKGROUND: Quantitative analysis of T-lymphocyte subsets is used to assess immune competency. Traditionally, T-lymphocyte subset analysis has been performed using flow cytometry, which requires complex instrumentation and relatively skilled manual operation. We evaluated the performance of an automated haematology analyser, the CELL-DYN Sapphire (CD Sapphire; Abbott Laboratories, USA) for T-lymphocyte subset analysis. METHODS: The precision and linearity obtained using the CD Sapphire was evaluated. T-lymphocyte subsets in blood samples from 120 patients were quantified using CD Sapphire and flow cytometry (Cytomics FC 500; Beckman-Coulter, France). The time required for complete T cell subset analysis using both methods was also evaluated. RESULTS: Results of CD Sapphire-based quantitation of CD3+, CD3+CD4+, and CD3+CD8+ cells showed intra-assay CV of less than 5% for precision and displayed linearity in the ranges of 84 to 5364, 41 to 2615, and 44 to 2800 cells/microL, respectively. There was good correlation among the CD3+, CD3+CD4+, and CD3+CD8+ cell counts as well as in the CD4/CD8 ratio (r=0.987, 0.982, 0.982, and 0.980, respectively) using CD Sapphire and flow cytometry. The mean turnaround time for the CD Sapphire (10.0+/-0.5 minutes) was significantly less than that for flow cytometry (111.8+/-8.4 minutes, P<0.001). CONCLUSIONS: T cell subset analysis using the CD Sapphire gives excellent performance and consistent results that correlate well with those obtained by flow cytometry. We conclude that this time-efficient method can replace conventional flow cytometric methods used for measuring T cell subsets.
Aluminum Oxide* ; CD4-Positive T-Lymphocytes ; CD8-Positive T-Lymphocytes ; Cell Count ; Flow Cytometry ; Humans ; T-Lymphocyte Subsets*

CD4+ CD25+ regulatory T cells (Treg) play a fundamental role in the establishment and maintenance of immune tolerance. In a some of experimental models, it was found that Tregs can quench autoimmune diseases, maintain allogeneic transplants, and prevent allergic diseases. A major obstacle to their clinical application is related to their definitive phenotype and very limited number of these cells in peripheral circulation, no more than 5%-10% of total CD4+ T cells. Recent progress of technologies for Treg sorting with multicolor flow cytometry and immuno-absorbing columns has overcome these obstacles, and opened the doors to the clinical application of Treg. This review highlight the characteristics of Treg, describe the current information of cell sorting and ex vivo expansion techniques, and outline the adoptive transfer experiments and clinical trials of immunotherapy that have been developed in recent years. It is foreseeable that Treg adoptive transfusion will be a promising immunosuppressive therapy.
Autoimmune Diseases ; CD4-Positive T-Lymphocytes ; Flow Cytometry ; Humans ; Immune Tolerance ; Immunotherapy ; T-Lymphocytes, Regulatory

OBJECTIVES: Chronic asthma has a number of characteristic feature; the increased airway responsiveness and bronchial inflammation. Although these mechanisms are not clear, activated T-cell has had an important role in migration and activation of inflammatory cells. In order to evaluate the role of T-lymphocyte and T-cell subsets in the development of asthmatic symptoms and the posibility for predicting the therapeutic response, we performed bronchoalveolar lavage from asymptomatic and symptomatic asthmatic subjects and inflammatory cell count, T-cell subset, activated T-lymphocyte were analysed and they were compared with healthy controls. METHOD: 76 bronchial asthmatics and 54 healthy controls were enrolled in this study. Asthmatic patients were classified into symptomatic and asymptomatic group according to symptom severity. Symptomatic group was divided into two groups according to therapeutic response ; early responder(ER) and late responder(LR). Lymphocytes(T-lymphocytes subsets and activation marker) in bronchoalveolar lavage(BAL) cells were analyzed using a flow-cytometry. RESULTS: The counts of eosinophil and neutrophil in BAL fluid were significantly higher in both asymptomatic and symptomatic asthmatic patient than those of healthy controls (p<0.05). The number of T3, T4, and T8 lymphocytes were significantly higher in symptomatic asthmatic patient than those of healthy controls, and the counts of T3-IL2R+, T4-IL2R+, and T8-IL3R+ lymphocytes were significantly higher in symptomatic subsets than in healthy controls(p<0.05). Although there were no significant differences in the number of lymphocyte, eosinophil and neutrophil between the ER and LR of symptomatic patients(p>0.05), the numbers of T3 and T4 lymphocyte subsets were significantly higher in LR than in healthy controls, and the number of T3-IL2R+, T4-IL2R+ lymphocytes were significantly higher in ER than in healthy controls(p<0.05). CONCLUSION: We could conclude that the infiltration and activation of T-lymphocytes might be associated with the development of asthmatic symptoms and responsiveness to therapy.
Asthma ; Bronchoalveolar Lavage Fluid* ; Bronchoalveolar Lavage* ; CD4-Positive T-Lymphocytes ; CD8-Positive T-Lymphocytes ; Cell Count ; Emigration and Immigration ; Eosinophils ; Humans ; Inflammation ; Lymphocytes ; Neutrophils ; T-Lymphocyte Subsets ; T-Lymphocytes*

The depressed cell-mediated immunity in pregnant women may play an important role inthe protection of the fetus from rejection by the mother. In this of view, many studieshave been performed to evaluated of lymphocyte subsets in recurrent spontaneous abortionwomen. However, because of the discordance in published data, auther have reevaluatedperipheral blood T-lymphocyte, B-lymphocyte and subsets defined by monoclonal antibodieswith immunobead method for 37 recurrent spontaneous abortion women and 30 normal healthywomen.The results were as follows. No significant difference was observed in the componentratio of the B cell. The component ratio of T lymphocyte in study group(70.5+/-10.1%) wassignificantly decreased in control group(79.7+/-3.1%)(p < 0.001). The component ratio of Nullcell in study group(19.0+/-10.1%) was higher than control group(9.4+/-2.6%)(p < 0.001). In thecomponent ratio of T4 cell, no statistically difference were observed in both groups. In thecomponent ratio of T8 cell, study group(28.4+/-9.5%) was significantly lower than controlgroup(38.4+/-3.2%)(p < 0.001). T4/ T8 ratio in the study group(1.58+/-0.70%)was significantly higher than control group(1.09+/-0.14%)(p < 0.001).In the component ratio of T/B cell, no statistically differences were observed.In conclusion, decreased T8 cell component ratio and increased T4/T8 component ratio may be play an important role in recurrent spontaneous abortion.
Abortion, Spontaneous* ; B-Lymphocytes* ; CD4-Positive T-Lymphocytes ; CD8-Positive T-Lymphocytes ; Female ; Fetus ; Humans ; Immunity, Cellular ; Lymphocyte Subsets ; Lymphocytes ; Mothers ; Pregnancy ; Pregnant Women ; T-Lymphocytes*

To evaluate the separation of T lymphocyte subsets by immunomagnetic beads and to find optimization of strategy for specific binding of antibody-coated beads to cells, two strategies to isolate enriched T lymphocyte subpopulation CD4+ T cells and CD8+ T cells from small volumes (< 5 ml) of peripheral blood by using immunomagnetic beads or complement cytotoxicity method were compared. The purity and activity of CD4+ T cells and CD8+ T cells were measured by using flow cytometry, trypan-blue dye exclusion test, etc. The results showed that the yields of CD4+ T lymphocytes and CD8+ T lymphocytes by using immunomagnetic beads were (94.2 +/- 1.4)% and (93.8 +/- 3.0)% respectively, higher than those of control group and the group of using completement cytotoxicity method (P < 0.05). At the same time, the yields of CD4+ T lymphocytes and CD8+ T lymphocytes by using complement cytotoxicity method were (76.0 +/- 2.8)% and (77.0 +/- 3.0)% respectively, higher than those of unenriched group (P < 0.05). The trypan-blue dye exclusion test confirmed that there were no influences on activity of CD4+ T cells and CD8+ T cells when immunomagnetic beads were used for separation of these cells from peripheral blood. It is concluded that the immunomagnetic bead method has a higher efficiency for separation of CD4+ T cells and CD8+ T cells from peripheral blood than complement cytotoxic method, especially for small sample. This method has no influence on activity and proliferation of T lymphocyte subpopulations, and would be expected to establish conditions for research of biological characteristics of CD4+ T cells and CD8+ T cells in future.
CD4-Positive T-Lymphocytes ; cytology ; CD8-Positive T-Lymphocytes ; cytology ; Flow Cytometry ; Humans ; Immunomagnetic Separation ; methods ; Leukocytes, Mononuclear ; cytology ; immunology

Humans ; CD4-Positive T-Lymphocytes/metabolism* ; CD8-Positive T-Lymphocytes/metabolism* ; HIV Infections/drug therapy* ; HIV-1

OBJECTIVETo investigate the clinical significance of memory T cells (CD45RO⁺ T) and the initial T cells (CD45RA⁺ T) distribution in peripheral blood of patients with peripheral T-cell lymphoma (PTCL).

METHODSA total of 27 patients diagnosed as PTCL in our hospital from February 2010 to February 2014 were collected in this study; whereas 30 healthy people were enrolled as control. The distribution of CD45RO⁺ T and CD45RA⁺ T cells were detected seperately in each group, and the results were analysed further.

RESULTSThe expression of T cell antigens in lymphnodes of PTCL patients were diverse: the CD4⁺ T cells were the main immune phenotype, while the B cell-related antigen was not expressed. The CD4⁺/CD8⁺, CD4⁺ CD45RO⁺ T in the peripheral blood of PTCL patients were significantly lower than that in normal group (P < 0.05); while the CD4, CD45RA⁺ T, CD8⁺ CD45RA⁺ T and CD8⁺ CD45RO⁺ T were significantly higher than that in normal group (P < 0.05). The patients in stage I/II had higher CD4⁺/CD8⁺, CD4⁺ CD45RO⁺ T than those in stage III/IV (P < 0.05), whereas the CD4⁺ CD45RA⁺ T, CD8⁺ CD45RA⁺ T and CD8⁺ CD45RO⁺ T were significantly lower than those in the stage III/IV patients (P < 0.05).

CONCLUSIONThe distributions of CD45RA⁺ T and CD45RO⁺ T in PTCL patients are quite different, and the corresponding treatment might be adopted according to the different distribution of these cells, so as to improve the diagnosis and prognosis of PTCL.