Common variable immunodeficiency disorders (CVID) are a group of heterogeneous conditions that have in common main failure of B cell function, although numerous T cell abnormalities have been described, including reduced proliferative response and reduced regulatory T cells. Putative follicular T cells, recent thymic emigrants and regulatory T cells were also assessed. Significant reduction in naive CD4 T cells, with reduced total CD4 and recent thymic emigrant figures, was observed in CVID patients, most pronounced in those with autoimmune cytopenias or polyclonal lymphoproliferation. These findings suggest a lack of replenishment by new thymically produced cells. CD8 naive T cells were reduced in CVID patients, most significantly in the autoimmune cytopenia subgroup. There was a reduction in early differentiated CD4 and CD8 T cells and increased CD8 TEM in the CVID patients, particularly autoimmune cytopenia and polyclonal lymphoproliferation subgroups, suggesting a more activated T cell phenotype, due perhaps to an antigen-driven process. XLA patients experienced significantly reduced putative follicular T cells, which may depend on W cells for survival, while no significant modifications were observed in the T cells of those with IgG subclass deficiency or selective IgA deficiency. W were performed, and if absent/low responses were noted the patient was vaccinated and these retested after 1 month. Lymphocyte subsets, both percentage and complete count, were also performed, including measurement of W cells, CD4 and CD8 T cells and natural monster (NK) cells [3,27]. At the time of analysis, all XLA and 55 of 58 CVID patients were on immunoglobulin replacement, but not on immunosuppressive therapy. Hederagenin supplier Those with autoimmune cytopenia or lymphoid interstitial pneumonia experienced not received corticosteroid therapy within 6 months, and only at prior doses <25 mg/kg. Hederagenin supplier No individual experienced an affected parent, sibling or child. CVID patients were categorized into the following clinical phenotypes, as explained in Chapel = 5), psoriasis (= 6), uveitis (= 2), vitiligo (= 2), pernicious anaemia (= 3), ulcerative colitis (= 4) and type 1 diabetes (= 2). Only one patient experienced a subsequent lymphoid malignancy and only three experienced an enteropathy, so these groups were not utilized in the analysis; these patients were included in the CVID total group. Physique 1 demonstrates the distribution of clinical phenotypes of the CVID patient group. Fig. 1 Venn NOTCH2 diagram illustrating the distribution of common variable immunodeficiency disorder (CVID) patients into clinical phenotypes [2,3]. Figures in brackets show figures in each division; patients may appear Hederagenin supplier in more than one group, as indicated. At the: … The number of patients stated in each group in Table 1 is usually the maximum number of patients analysed for a T cell subpopulation. However, for some of the T cell subpopulations smaller figures were analysed due to either technical troubles with a particular tube or limited sample availability. Circulation cytometry All circulation cytometric analysis was performed on ethylenediamine tetraacetic acid (EDTA) blood samples within 48 h of venepuncture. Lymphocyte subset analysis and complete counts of total lymphocytes, total T cells, CD4 and CD8 T, W and NK cells were performed using BD Multitest? CD3/CD16+CD56/CD45/CD19 and CD3/CD8/CD45/CD4 with BD Trucount? Tubes (Becton-Dickinson, San Jose, CA, USA) and acquired on a BD fluorescence activated cell sorter (FACS)Calibur (Becton Dickinson), as per the manufacturer’s instructions. For T cell subpopulations, 100 t of whole blood was incubated with directly conjugated fluorescent antibodies for 30 min in the dark at room heat, then reddish cells were lysed using FACSlyse (Becton-Dickinson), washed in phosphate-buffered saline Hederagenin supplier (PBS) and fixed in PBS with 1% formaldehyde. Samples were acquired using four-colour purchase on a FACSCalibur and data analysed using CellquestPro software (Becton-Dickinson). Fluorescence minus one gating techniques were employed to evaluated thresholds for positivity of individual antibodies and aid gating of T cell subpopulations. The following CD3+ T cell subpopulations were analysed on CD4 and CD8 cells: naive, central memory (CM), effector memory (EM) and terminally differentiated decided (TEM) by CCR7 and CD45RA manifestation; early, intermediate and late differentiation status was decided by CD28/CD27 manifestation. Other CD4 T cell populations included recent thymic emigrant (defined by CD45RA/CD31), putative follicular T cells (defined by CXCR5/CD45RO) and Tregs (defined by CD25+CD127-). Complete cell counts were calculated using the CD4 or CD8 T cell counts from the lymphocyte subset analysis. Subpopulations were added together to ensure that the total number of CD4 or CD8 matched up those from the lymphocyte subset analysis. Statistical analyses All statistical analyses were performed using GraphPad Prism version 4 (GraphPad Software, San Diego, CA, USA). All data were analysed using non-parametric one-way analysis of variance (anova) KruskalCWallis with Dunn’s multiple comparison test as a test or one-way anova with Tukey’s multiple comparison test as a test. T cell subpopulation correlations with age were analysed by Spearman’s correlation. The Venn diagram (Fig. 1) was made using.