Simplified assay for enrichment of primed human Th17 and Tc17 lymphocytes from peripheral blood

Translational Medicine Communications

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[1] 0000 0001 2297 5165, grid.94365.3d, Flow Cytometry Core Facility, National Heart Lung and Blood Institute, National Institutes of Health, 10 Center Dr., Bldg. 10, Rm 8C104, 20892, Bethesda, Maryland, USA;Lab Head Immune Profiling, Precision Immunology, Immunology & Inflammation Research Therapeutic Area, 270 Albany Street, Office# 2212-A, 02139, Cambridge, MA, USA;

关键词: MCAM;    CD146;    Th17;    Tc17;    Flow cytometry;   

DOI  :  10.1186/s41231-019-0041-8

来源: publisher

【 摘 要 】

BackgroundInterleukin-17A (IL-17A) is a potent pro-inflammatory cytokine that has been implicated in the pathogenesis of various autoimmune diseases. The production of IL-17A is commonly associated with subsets of CD4+ T cells (Th17) and CD8+ T cells (Tc17). Identifying these subsets based on intracellular expression of IL-17 or transcription factor RORC precludes isolation of viable Th17 and Tc17 cells and there by limits studies involving cell-cell interaction or cellular functions. Therefore, identifying surface markers that can help in identifying and enriching these cells is important.ResultsWe used MCAM as a surrogate marker to identify in vivo committed human Th17 and Tc17 subsets. By employing high-speed fluorescence activated cell sorting, we enriched IL-17A-producing subsets from human specimens without the need for in vitro polarization using exogenous cytokines. These subsets can be investigated, following sorting, using a variety of methods such as ELISA, ex vivo functional assays and next generation sequencing to gain insights into the role of human Th17 and Tc17 in health and disease.ConclusionWe here demonstrate that both CD4+ T cells (Th17) and CD8+ T cells (Tc17) cell populations can be identified based on the surface expression of melanoma cell adhesion molecule (MCAM or CD146).

【 授权许可】

CC BY