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. 2020 Oct 29;1(3):100154. doi: 10.1016/j.xpro.2020.100154

Figure 6.

Figure 6

Example of Gating Strategy for the Characterization of Th Subsets in Memory CD4+ T Cells

Representative example of analyses in a cryopreserved PBMC sample. Memory CD4+ T cells are first selected by excluding naive cells (CCR7+ CD45RA+ CD4+ T cells). Inside memory CD4+ T cells, SLeX+ were selected from Stain 1, using the antibody cocktail described on Table 3. CLA+ cells (black gate), CLAhi (teal gate), and CLAlow (orange gate) were selected from Stain 2, using the antibody cocktail described in Table 4. cTfh cells are identified using PD-1 and CXCR5. CD127low CD25+ cells were selected within non-cTfh cells, and Foxp3+ cells were then selected to identify Tregs. Non-cTfh/Tregs cells were selected based on CXCR5 and CCR6 expression. Th1 and Th2 cells were identified within the CXCR5- CCR6- gate, while Th17 and Th17+1 were characterized ins the CXCR5- CCR6+ gate using CXCR3. The frequency of CXCR3+ Tfh can also be measured with this panel.