Project description:Gene-environment interactions are implicated in immunopathology, but few specific mechanisms have been identified. We defined two terminal-differentiation pathways for human CD4+ T cells each marked by CD57. Rare blood CD57+ CD4+ T cells marked by TCF1 downregulation (Thcyt) adopt a cytotoxic and terminal-effector transcriptome. By contrast, human CD57+ GC-Tfh cells exhibit a transcriptome of the precursor exhausted T cells marked by TCF7, TOX and ID3 and constitutive expression of CTLA4, which collectively confer resistance to becoming cytotoxic. By clarifying human CD4+ T cell terminal differentiation, we have revealed how CTLA4 mitigates the risk of maladaptive cytotoxicity during infection.
Project description:We have identified a CD57+PD1- CD4 T cell phenotype at the time of transplantation that strongly correlates with subsequesnt development of belatacept-resistant rejection. In this study, we used microarray to determine which genes were upregulated in CD57+ compared to CD57- CD4 T cells. Peripheral blood obtained from 5 healthy controls was processed and sorted into CD4+CD57+PD1- and CD4+CD57-PD1- populations. Total RNA was extracted from the sorted populations and quality assessed. cDNA synthesis and amplification was performed, and fragmented and biotinylated samples were hybridized to the Affymetrix Human U133 plus 2.0 probe array. The arrays were scanned and probe intensity measurements were normalized across the samples using the robust multichip average (RMA) algorithm.
Project description:We have identified a CD57+PD1- CD4 T cell phenotype at the time of transplantation that strongly correlates with subsequesnt development of belatacept-resistant rejection. In this study, we used microarray to determine which genes were upregulated in CD57+ compared to CD57- CD4 T cells.
Project description:We used microarrays to compare the gene expression profile of DP and DN (KLRG1 and CD57) CD4+ T cells from luminal breast cancer patients and healthy donors.
Project description:Type 1 diabetes is an autoimmune disease in which insulin-secreting β cells of the pancreatic islets are selectively destroyed. CD8 T cells are regarded as critical players in mediating β cell destruction and as a result, considerable effort has been expended to define CD8 T cell behaviour in this disease. The overarching aim of the experiment is to characterize a recently identified autoreactive CD57-positive CD8 T cell subset which is associated with loss of function of islet beta cells in type 1 diabetes, to compare the phenotype of the CD57-positive effector memory CD8 T cells versus the CD57-negative compartment, and provide an insight into the function of these cells in the disease process. To that aim, HLA-A*24 positive patients with T1D -within 2 years of diagnosis- were asked to provide a blood, and following consent, and PBMC were isolated. CD57-positive and CD57-negative CD8 T cell populations were sorted by FACS, and finally, RNA was extracted. Amplified cDNA was obtained and used for the library preparation.
Project description:In human studies, mononuclear cells from peripheral blood (PBMC) often serve as a source of clinical study markers. Therefore, it is essential to understand to what extent phenotypes of PBMCs reflect their tissue-resident equivalents. In order to determine the heterogeneity of T-follicular helper (TFH) cells in peripheral blood versus tonsils, CD3+CD4+CD45RA–CXCR5+ cells of both origins were sorted and transcriptomes, TCR repertoires and cell-surface protein expression were analysed by single-cell RNA sequencing, flow cytometry and immunohistochemistry. Four subsets of TFH cells with classical T-follicular gene expression patterns were mostly found in tonsils. Interestingly, however, all circulating CD3+CD4+CXCR5+ T-cell subpopulations also appeared in tonsils. Still, when circulating, cells of the same cluster rather displayed markers of proliferation and migration, whereas their tonsillar counterparts exhibited known TFH characteristics resembling bona fide germinal center-typical TFH cell subtypes. Surprisingly, one distinct and oligoclonal CD4+CXCR5+ subpopulation displayed pronounced cytotoxic properties. Those ‘killer TFH (TFK) cells’ were found among PBMCs as well as tonsillar cells but were located outside of germinal centers. Accordingly, they were the only CD4+CXCR5+ T-cell subtype in tonsils featuring transcripts for CXCR3 and PSLG1. They appeared terminally differentiated and could be distinguished from all other TFH subsets by expression of NKG7 (TIA-1), granzymes, perforin, CCL5, CCR5, EOMES, CRTAM and CX3CR1. This hitherto undescribed subpopulation is indicative for chronic infection and inflammation and therefore is a valuable candidate to be included in any CD4+CXCR5+ TFH cell assessment.
Project description:Gene Array Analysis of CD8+ CD57+ T cells in HIV Patients. Gene expression patterns of CD8+CD57+ T cells from healthy donors and HIV patients were compared. Gene expression patterns of CD8+CD57- T cells and CD8+CD57+ T cells were compared.