MBTP 29: Immunophenotyping of antigen-specific CD4+ T cells from human PBMCs using flow cytometry

Miltenyi Biotec-tested panel 29 (MBTP 29)

This application protocol describes the flow cytometric analysis of antigen-specific CD4+ T cells from human PBMCs.

Protocol

Gating strategy showing the analysis of antigen-specific CD4+ T cells from human PBMCs. Antigen-specific CD4+ T cells were stained using the previously described panel. Samples were initially gated on live cells using Viobility-negative gating (data not shown). Live cells were gated on CD3 expression to identify T cells (A). T cells were then further gated on expression of CD4 and the activation marker CD154. CD4+ CD154+ cells were considered ‘antigen-specific’. Antigen-specific CD185+ cells were further analyzed for the concomitant expression of CD183 (Follicular T helper cells 1 (TFH1)) and CD196 (Follicular T helper cells 17 (TFH17)) (C, D).

Materials

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