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Bulk RNA-sequencing of IL-21-tRFP+ CD44+ Th1, Tfh, and memory-like CD4+ T cell subsets on day 14 post LCMV Cl13 infection

ABSTRACT: CD4+ T cell-derived interleukin 21 (IL-21) sustains CD8+ T cell responses during chronic viral infection, but the helper subset that confers this protection remains unclear. Here, we applied scRNA and ATAC-seq approaches to determine the heterogeneity of IL-21+CD4+ T cells during LCMV clone 13 infection.

ORGANISM(S): Mus musculus

PROVIDER: GSE193934 | GEO | 2022/02/25

REPOSITORIES: GEO

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ATAC-sequencing of Th1, Tfh, and memory-like splenic CD4+ T cells on day 14 post LCMV Cl13 infection

Project description:CD4+ T cell-derived interleukin 21 (IL-21) sustains CD8+ T cell responses during chronic viral infection, but the helper subset that confers this protection remains unclear. Here, we applied scRNA and ATAC-seq approaches to determine the heterogeneity of CD4+ T cells during LCMV clone 13 infection.

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Single-cell RNA-sequencing of IL-21-tRFP+ GP66+ splenic CD4+ T cells on days 7 and 14 post LCMV Cl13 infection

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Bulk RNA-sequencing of IL-21-tRFP+ CD44+ Th1, Tfh, and memory-like CD4+ T cell subsets on day 14 post LCMV Cl13 infection

Project description:Bulk RNA-sequencing of IL-21-tRFP+ CD44+ Th1, Tfh, and memory-like CD4+ T cell subsets on day 14 post LCMV Cl13 infection

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RNA-seq of Tfh IL21+ single producers and Tfh IL21+ IL10+ double producers

Project description:During chronic viral infection, the inflammatory function of CD4 T cells becomes gradually attenuated. Concurrently, Th1 cells progressively acquire the capacity to secrete the cytokine IL-10, a potent suppressor of antiviral T cell responses. To determine the transcriptional changes that underlie this T cell adaption process, we applied a single-cell RNA-sequencing approach and assessed the heterogeneity of IL-10-expressing CD4 T cells during chronic infection. Unexpectedly, our analyses revealed an IL-10-producing population with a robust Tfh-signature. Using IL-10 and IL-21 double-reporter mice, we further demonstrate that IL-10+IL-21+co-producing Tfh cells arise predominantly during chronic but not acute LCMV infection. Importantly, depletion of IL-10+IL-21+co-producing CD4 T cells or deletion of Il10 specifically in Tfh cells resulted in impaired humoral immunity and viral control. Mechanistically, B cell-intrinsic IL-10 signaling was required for sustaining germinal center reactions. Lastly, we demonstrate that IL-27 and type I IFNs differentially regulate the formation of this protective IL-10-producing Tfh subset. Thus, our findings elucidate a critical role for Tfh-derived IL-10 in promoting humoral immunity during persistent viral infection.

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

Project description:CD4 T cell help is critical for both the generation and maintenance of germinal centers, and T follicular helper (TFH) cells are the CD4 T cell subset required for this process. SAP (SH2D1A) expression in CD4 T cells is essential for germinal center development. However, SAP-deficient mice have only a moderate defect in TFH differentiation as defined by common TFH surface markers. CXCR5+ TFH cells are found within the germinal center as well as along the boundary regions of T/B cell zones. Here we show that germinal center associated T cells (GC TFH) can be identified by their co-expression of CXCR5 and the GL7 epitope, allowing for phenotypic and functional analysis of TFH and GC TFH populations. Here we show GC TFH are a functionally discrete subset of further polarized TFH cells, with enhanced B cell help capacity and a specialized ability to produce IL-4 in a TH2-independent manner. Strikingly, SAP-deficient mice have an absence of the GC TFH subset and SAP- TFH are defective in IL-4 and IL-21 production. We further demonstrate that SLAM (Slamf1, CD150), a surface receptor that utilizes SAP signaling, is specifically required for IL-4 production by GC TFH. GC TFH cells require IL-4 and IL-21 production for optimal help to B cells. These data illustrate complexities of SAP-dependent SLAM family receptor signaling, revealing a prominent role for SLAM receptor ligation in IL-4 production by germinal center CD4 T cells but not in TFH and GC TFH differentiation. Analysis of in vivo polyclonal GC Tfh vs Tfh vs Non-Tfh eight days after LCMV viral infection. Analysis of in vivo follicular helper CD4 T cells (CXCR5high GL7low), versus germinal center follicular helper CD4 T cells (CXCR5hi GL7hi), versus non-follicular helper CD4 T cells (CXCR5low) eight days after viral infection.

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Expression data from virus-specific mouse Th1 and Tfh CD4 T cell subsets after LCMV infection

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The role of IL-6 and IL-27 signalling on virus specific CD4 T cell responses during LCMV Cl13 infection

Project description:LCMV Clone 13 is an arenavirus that results in a persistent viral infection in mice when delivered intravenously. It has been widely used to explore the impact of chronic viral infection on CD4 T cell responses and dissect mechanisms that regulate this. In this study we used mixed bone marrow chimeras from wildtype and cytokine reeptor deficient (gp130 or IL-6/IL-27R) mice to determine the role these cytokines on splenic CD4 T cell responses 30 days post infection

2023-07-15 | GSE235035 | GEO

Single-cell RNA-sequencing of CD44+ CD4+ T cells from day 21 post-LCMV Cl13 infection following transient CD4+ T cell depletion (scRNA-Seq)

Project description:CD4+ T cells play a critical role in sustaining the effector function of CD8+ T cells during chronic viral infection. When CD4+ T cell “help” is absent, following transient CD4+ T cell depletion, CD8+ T cells enter a dysfunctional state, losing their capacity for viral control. Here, we applied scRNA-seq to determine the CD4+ T cell subsets that repopulate following transient CD4+ T cell depletion, during LCMV Clone 13 infection.

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Single cell RNA-seq of IL-10-producing CD4 T cells during chronic LCMV infection

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2022-07-01 | GSE175741 | GEO

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