Figure 1.

Gating strategy to identify Th populations among T-effector cells. Shown is a representative sarcoidosis subject sample. We gated on singlet cells using FSC-H and FSC-A, then live cells (negative for the fixed viability dye), lymphocytes based on FSC and SSC, then CD3⁺ and CD4⁺ cells. We then gated on NonTregs that were CD25⁻and either CD127^Lo or CD127^Hi and then T-effectors (CD45RA⁻ and CD45RO⁺). Among these T-effectors, we enriched for Th subsets using CCR6, CCR4, and CXCR3. Th17.0 cells were CCR6⁺CCR4⁺CXCR3⁻, Th17.1 cells were CCR4⁻CCR6⁺CXCR3⁺ and Th1 cells were CCR6⁻CCR4⁻CXCR3⁺. The gating strategy used for fluorescence minus one (FMO) controls for CCR6, CCR4, and CXCR3 are shown in the lower left corner.