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. 2015 Sep 3;9(9):e0004067. doi: 10.1371/journal.pntd.0004067

Fig 1. Flow cytometry gating strategy for Th subsets isolated from (A) the blood and (B) the foreskin.

Fig 1

Blood cells were isolated by density centrifugation, and foreskin cells by mechanical and enzymatic digestion of tissue. Dead cells and doublets were excluded. Lymphocytes were identified based on characteristic size and granularity in blood samples and gates applied to foreskin samples. CD4 T cells were identified by expression of CD3 and CD4. Th subsets among CD4 T cells were identified by cytokine production in response to non-specific stimuli (SEB shown). Gates for cytokine production were based on unstimulated samples. Th17 cells were identified by production of IL-17A; Th1 cells by production of IFNγ and not IL-17A; Th22 cells by production of IL-22 in the absence of either IFNγ or IL-17A. Representative plots showing expression of CCR5, CD69 and HLA-DR on CD4 T cells are also shown.