Gating scheme for the analyses of the different T cell subsets in the peripheral blood mononuclear cells (PBMC) from a representative animal.

The lymphocytes were first gated based on forward scatter (FCS) versus side scatter (SSC), and then live lymphocytes were identified based on SSC and aqua-negative population. The T cells were then positively identified by CD3 expression followed by the detection of the CD4⁺ CD8⁻ (CD4⁺ T cells) and CD4⁻ CD8⁺ (CD8⁺ T cells) populations within the CD3⁺ T cells. On the basis of CD28 and CD95 expression, the CD4⁺ and CD8⁺ T cells were further differentiated into naïve (Tn CD28⁺ CD95⁻), central memory (Tcm CD28⁺ CD95⁺) and effector memory (Tem CD28⁻ CD95⁺) subsets.