Figure 5. TDL CXCR5^BrPD-1^Br Tfh are transcriptionally similar to GC Tfh.

(A) Gating strategy for sort purification with an overlay of flow cytometry plots for blood (n = 3), TDL (n = 3), and mesenteric LNs (n = 3): naive (orange gate) and nonnaive (black gate). Nonnaive cells were sorted based on the gates shown for each tissue source: CXCR5^BrPD-1^Br (red), CXCR5^DimPD-1^Dim (teal), CXCR5^– non-Tfh (green) and cTfh (blue). Plots for TDL and blood are shown in Figure 1A. mLN, mesenteric lymph node. (B) Principal component analysis of RNA-Seq data; colors represent populations from the matching gate color in A. (C) Correlation plot of the fold change from CXCR5^BrPD-1^Br Tfh to CXCR5^DimPD-1^Dim Tfh in TDL (TDL^Br and TDL^Dim, respectively) and LNs (GC Tfh and non-GC Tfh, respectively). (D) GSEA of transcriptional signatures from tonsil GC Tfh versus tonsil non-GC Tfh (GSE50391) compared with differentially expressed genes in GC versus non-GC Tfh from LNs and (E) TDL^Br Tfh versus TDL^Dim Tfh. Normalized enrichment scores (NES) and FDR marked on each GSEA. (F) Heatmap of leading edge genes from the LNs and TDL GSEAs in D and E were compared for overlap in both positively and negatively enriching genes. (G) Volcano plot of differentially expressed transcripts (adjusted P < 0.05 in orange) for LN Tfh (dim and bright) versus all TDL Tfh (dim and bright). (H) Volcano plot for differentially expressed transcripts (adjusted P < 0.05 in orange) for all TDL Tfh (dim and bright) versus blood cTfh.