Fig. 7: Cell surface proteins identify distinct human tonsillar T_FH and cT_FH phenotypes.

a–e, Flow cytometry analysis of CD3⁺CD4⁺CD45RA⁻CD45RO⁺CXCR5⁺ T_FH cells from five PBMC and six tonsil healthy adult donors. t-distributed stochastic neighbor embedding (t-SNE) and heatmap expression of T_FH cell markers of PD-1, CXCR3, CCR6 and CCR4 (a). T_FH cell populations identified by PD-1 and CD57 with expression of the T_FH cell markers CXCR5, ICOS, OX40 and CCR4 on each population (b). Histograms of PD-1, CD57, CD69 and CD82 expression on T_FH cells (c). Histograms of CD127, CD99, CD71 and CD151 expression on T_FH cells (d). T_FH populations separated into quadrants (Q1–Q4) based on expression of CD99 and CD151 markers (e). f, scCITEseq surface protein expression (log counts) of sorted CD3⁺CD4⁺CD45RA⁻CD45RO⁺CXCR5⁺CD27⁺ human tonsillar T_FH cells from three healthy adult donors (as in a–e) of cluster-identifying markers overlaid onto the scRNA-seq UMAP. g, T_FH cell surface and activation markers in CD3⁺CD4⁺CD45RA⁻CD45RO⁺CXCR5⁺ human T_FH cells from matched tonsil, adenoid tissue and PBMC from five healthy juvenile donors. h,i, SARS-CoV-2 Spike tetramer⁺ CD3⁺CD4⁺CD45RA⁻CXCR5⁺ cT_FH cells from SARS-CoV-2 infected donors (h) at time of infection (11 of 11 donors; n = 11) and 6 months convalescence (10 of 11 donors; n = 10) and nine COVID-19 mRNA vaccinated donors (n = 9) (i) at 7 days post-vaccine and 3 months post-vaccine. Frequency of Spike tetramer⁺PD-1⁺ T_FH cells, gMFI expression for CXCR3 and CCR6, and frequencies and dot plots of CD151/CD99 quadrant-separated T_FH cell populations. Data show experiments of 8–11 samples per group. Statistical test was a two-tailed paired Student’s t-test. P values are indicated.

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