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Stepwise differentiation of follicular helper T cells reveals distinct developmental and functional states [scRNA-seq]

ABSTRACT: Follicular helper T (Tfh) cells are essential for the formation of high affinity antibodies. However, the signals that promote Tfh development after initial differentiation are unknown. Here we use fate mapping and reporter strategies to show that, after initial differentiation, Tfh cells transition from a progenitor-like Tfh (Tfh-Prog) stage to an IL-21-expressing fully developed Tfh (Tfh-Full) stage which is regulated by follicular regulatory T (Tfr) cells. Using single cell RNA sequencing and epigenomic analysis, we uncover additional developmental stages including an early Tfh transitory (Tfh-Trans) stage prior to full development, as well as an Tfh-Ex stage that occurs after full development marked by loss of IL-21 expression. The transcriptional and epigenetic landscape is formed in the Tfh-Transitory stage and is maintained throughout the Tfh-Ex state, suggesting terminal development. We also find the transcription factor Foxp1 regulates the transitions between these stages in Tfh cells to balance the ratio of stem-like progenitor and effector Tfh cells. Through selective in vivo deletion of Tfh-Full and Tfh-Ex cells, we show that these cells control antigen-specific germinal center formation, maintenance and somatic hypermutation at distinct stages of responses to a SARS-CoV-2 vaccine. Together, these studies demonstrate the transitionary phases of Tfh development and how control of the progression through these stages regulates humoral immunity.

ORGANISM(S): Mus musculus

PROVIDER: GSE236876 | GEO | 2023/10/20

REPOSITORIES: GEO

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Stepwise differentiation of follicular helper T cells reveals distinct developmental and functional states [ATAC-Seq]

Project description:Follicular helper T (Tfh) cells are essential for the formation of high affinity antibodies. However, the signals that promote Tfh development after initial differentiation are unknown. Here we use fate mapping and reporter strategies to show that, after initial differentiation, Tfh cells transition from a progenitor-like Tfh (Tfh-Prog) stage to an IL-21-expressing fully developed Tfh (Tfh-Full) stage which is regulated by follicular regulatory T (Tfr) cells. Using single cell RNA sequencing and epigenomic analysis, we uncover additional developmental stages including an early Tfh transitory (Tfh-Trans) stage prior to full development, as well as an Tfh-Ex stage that occurs after full development marked by loss of IL-21 expression. The transcriptional and epigenetic landscape is formed in the Tfh-Transitory stage and is maintained throughout the Tfh-Ex state, suggesting terminal development. We also find the transcription factor Foxp1 regulates the transitions between these stages in Tfh cells to balance the ratio of stem-like progenitor and effector Tfh cells. Through selective in vivo deletion of Tfh-Full and Tfh-Ex cells, we show that these cells control antigen-specific germinal center formation, maintenance and somatic hypermutation at distinct stages of responses to a SARS-CoV-2 vaccine. Together, these studies demonstrate the transitionary phases of Tfh development and how control of the progression through these stages regulates humoral immunity.

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Project description:CD4+ T follicular helper (Tfh) cells are essential for germinal center (GC) and high-affinity antibody responses. Yet the regulation that determines the initial development of Tfh cells is still largely unknown. Here we find that transcription factor Foxp1, previously shown to be essential in maintaining T cell quiescence, is a rate-limiting and essential negative regulator of Tfh cell differentiation. NaM-CM-/ve CD4+ T cells constitutively express Foxp1A, and stimulation through the T cell receptor (TCR) transiently induces the expression of a shorter Foxp1D isoform. In T cell-dependent (TD) humoral responses, CD4+ T cells deficient in all Foxp1 isoforms preferentially differentiate into Tfh cells, resulting in substantially increased GC and antibody responses. This negative regulation of Foxp1 on Tfh cell differentiation shows profound dominance: even in the absence of B cells, Foxp1-deficient CD4+ T cells differentiate into Tfh cells with high frequencies and sustained Bcl6 expression. Further, in the absence of Foxp1, Tfh cells are generated in higher frequencies than seen with Bcl6 overexpression and Tfh cell differentiation becomes significantly resistant to Blimp1-mediated repression. Finally, our experiments reveal specific roles of Foxp1A and Foxp1D in inhibiting Tfh cell differentiation: TCR-induced Foxp1D functions as a M-bM-^@M-^XgatekeeperM-bM-^@M-^Y to block the initial Tfh cell development, and together Foxp1A and Foxp1D proteins inversely determine Tfh cell generation in a dosage-dependent manner. Our study suggests that two Foxp1 isoforms provide a M-bM-^@M-^\double checkM-bM-^@M-^] mechanism as fundamental regulators in Tfh cell differentiation and humoral responses. Gene expression analysis of ex vivo OT-II Foxp1-WT Tfh cells and OT-II Foxp-conditional knockout (CKO) Tfh cells 5 days after immunization.

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2014-01-01 | E-GEOD-52839 | biostudies-arrayexpress

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Expression Data from in vivo Tfh vs GC Tfh vs non-Tfh

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