Figure 1.

Gating Strategy for Tfh in Whole Blood

Gating strategy to identify cTfH, activation, and subsets. Whole blood was stained and analyzed by flow cytometry, and CD45⁺ lymphocytes were gated as T cells on CD3. CD4 T cells were gated as CD4⁺CD8⁻ and cTfh analyzed based on PD1⁺CXCR5⁺ cells. cTfh cell subsets were analyzed based on CXCR3 and CCR6 staining into Th1 (CXCR3⁺CCR6⁻), Th17 (CXC3⁻CCR6⁺), and Th2 (CXCR3⁻CCR6⁻). Activation was gated as ICOS⁺ and CD38⁺ “and” gates were made to assess activation markers within subsets. Data are representative of 157 samples (40 individuals at days 0, 8, and 14/15 and 37 individuals at EOS).