Project description:To determine the clinical relevance of the murine Th17-derived Tfh signatures in RA patients, we generated both splenic and PP murine Th17-derived Tfh cells signatures using bulk RNA-seq data.
Project description:Naive murine CD4+ T cells from GREAT/SMART-17A mice were cultured under Th1 or Tfh(1 ng/ml TGF-β)-polarizing conditions in 96-well plates coated with anti-CD3/anti-CD28 for 3.5 days; sorted by flow cytometry on IFNg+ (Th1), or CXCR5-IL17A+ (Th17) and CXCR5+IL17A- (Tfh); and subjected to bulk RNA-seq.
Project description:This experiment was designed to study the functional differences between the Th17-derived Tfh and regular Tfh cells. To achieve these goals, we performed flow sorting and enriched CD4+ T cells from Current-Ex Th17 fate-mapping mice. We then performed scRNA seq analysis to compare the transcriptomic profiles of Tfh cells.
Project description:This experiment was designed to study the functional differences between the Th17-derived Tfh and regular Tfh cells. We were also interested in studying how B cells might display functional differences upon receiving help from Th17-derived Tfh cells vs. regular Tfh cells. To achieve these goals, we performed flow sorting and enriched T-B cell doublets with either Th17-derived Tfh or regular Tfh cells to catch the T-B cell interaction “in action”. We then broke the doublets apart with ETDA before scRNA seq analysis to compare the transcriptomic profiles from both T cell side and B cells side.
Project description:We have shown that TGFb in combination with IL-6 potently induces a T follicular helper cell (Tfh) cell phenotype in activated murine CD4+ T cells in vitro. The transcription factor c-Maf critically contributes to the differentiation of Tfh cells in vitro and in vivo.
Project description:After activation, CD4+ helper T (Th) cells differentiate; into distinct effector subsets. Although chemokine; (C-X-C motif) receptor 5-expressing T follicular; helper (Tfh) cells are important in humoral immunity,; their developmental regulation is unclear. Here we; show that Tfh cells had a distinct gene expression; profile and developed in vivo independently of the; Th1 or Th2 cell lineages. Tfh cell generation was regulated; by ICOS ligand (ICOSL) expressed on B cells; and was dependent on interleukin-21 (IL-21), IL-6,; and signal transducer and activator of transcription; 3. However, unlike Th17 cells, differentiation of Tfh; cells did not require transforming growth factor; b (TGF-b) or Th17-specific orphan nuclear receptors; RORa and RORg in vivo. Finally, naive T cells activated; in vitro in the presence of IL-21 but not; TGF-b signaling preferentially acquired Tfh gene; expression and promoted germinal-center reactions; in vivo. This study thus demonstrates that Tfh is a; distinct Th cell lineage. Experiment Overall Design: Splenic CD4+CXCR5+ T cells were isolated from KLH-immunized mice and restimulated with anti-CD3 for 4 hours before total RNA preparation. Affymetrix gene chips were used to analyze their gene expression.
Project description:After activation, CD4+ helper T (Th) cells differentiate into distinct effector subsets. Although chemokine (C-X-C motif) receptor 5-expressing T follicular helper (Tfh) cells are important in humoral immunity, their developmental regulation is unclear. Here we show that Tfh cells had a distinct gene expression profile and developed in vivo independently of the Th1 or Th2 cell lineages. Tfh cell generation was regulated by ICOS ligand (ICOSL) expressed on B cells and was dependent on interleukin-21 (IL-21), IL-6, and signal transducer and activator of transcription 3. However, unlike Th17 cells, differentiation of Tfh cells did not require transforming growth factor b (TGF-b) or Th17-specific orphan nuclear receptors RORa and RORg in vivo. Finally, naive T cells activated in vitro in the presence of IL-21 but not TGF-b signaling preferentially acquired Tfh gene expression and promoted germinal-center reactions in vivo. This study thus demonstrates that Tfh is a distinct Th cell lineage.
Project description:In this study, the participation of fetal factors in maintaining the immune balance during gestation was investigated by assessing the effects of human chorionic gonadotropin (hCG) on murine Treg cells and Th17 cells.