Upregulation of CD9 identifies cytokine-producing T follicular helper (Tfh) cells during viral infection

Abstract

Abstract T follicular helper (Tfh) cells, a subset of CD4+ T cells, critically regulate the adaptive immune response by promoting germinal center (GC) B cell maturation into memory B cells and antibody secreting plasma cells. Tfh cells promote the GC response through the production of canonical cytokines IL-21 and IFNg, or IL-4 and CD40L signaling. Tfh cells progressively differentiate to regulate the germinal center response, but other than previous studies using cytokine reporter mice, the stages of this differentiation remain unclear. We identified that CD9, a tetraspanin protein, is temporally upregulated on a subset of Tfh cells following acute LCMV infection. To assess the function of the CD9 expressing Tfh cells, we compared migration to GC honing chemokines CXCL12 and CXCL13 and cytokine production between CD9hi and CD9lo Tfh cells at 8 days post LCMV infection. We found that CD9hi Tfh cells exhibited enhanced migration to CXCL12 and CXCL13 compared to CD9lo cells. Ex vivo stimulation of Tfh cells revealed that the CD9hi subsets were the primary producers of IL-21 and IFNg. Transcriptional analysis demonstrated that that CD9hi and CD9lo Tfh cells are distinct subsets characterized by enhanced expression of cell cycling genes in the CD9hi fraction. Thus, not only does CD9 identify the cytokine producing population of Tfh cells but marks a functionally and transcriptionally distinct subset promoting GC responses during viral infections.