Project description:T cells from spleen and lymphnode were isolated with EasySep Mouse CD8+ T Cell Isolation kit. Isolated CD8+ T cells were stimulated and cultured with plate-bound anti-CD3 and soluble anti CD28 antiboies in Tc1 or Tc9 cell polarizing conditios. After 3-day of polarization, cell were transferred into a new well and culture for another 2-day. on day 5 after seeding, Tc1 and Tc9 cells were collected and sequenced. We used microarrays to detail the global programme of gene expression of Tc1 and Tc9 cells at day 5 polarization.
Project description:The goal of this project is to dissect the mechanism by which Tc1 and Tc2 effector subtypes effect cell physiology in vitro. I utilized cellular proteomics to determine Tc1 and Tc2 secretion patterns and identify inflammatory factors that regulate cell polarization and physiology in vitro.
Project description:we use RNA sequence to detect the global gene expression in tumor-infiltrating Tc1 and Tc9 cells after being adoptively transferred in vivo
Project description:Type I immunity is defined by IFN-γ secretion, with Tc1 cells as a key subset, yet their intrinsic negative regulators remain unclear. We identified RASA3 as a novel negative regulator of Tc1 cells. T cell-specific deletion of RASA3 enhanced Tc1 differentiation and antibacterial immunity. We further revealed the RASA3–HCK–STAT4 axis in the regulatoin of Tc1 cell generation.
Project description:We provide an original multi-stage approach identifying a gene signature to assess the fibroblast polarization. Prototypic polarizations (inflammatory/fibrotic) were induced by seeded mouse embryonic fibroblasts (MEFs) with TNFα or TGFß1, respectively. The transcriptomic and proteomic profiles were obtained by RNA microarray and LC/MS-MS. Gene Ontology and pathways analysis were performed among the differentially expressed genes (DEGs) and proteins (DEPs). Balb/c mice underwent daily intradermal injections of HOCl (or PBS) as an experimental murine model of inflammation-mediated fibrosis in a time-dependent manner. As results, 1,456 and 2,215 DEGs, and 289 and 233 DEPs were respectively found in MEFs in response to TNFα or TGFß1, respectively. Among the most significant pathways, we combined 26 representative genes to encompass the proinflammatory and profibrotic polarizations of fibroblasts. Based on principal component analysis, this signature deciphered baseline state, proinflammatory polarization, and profibrotic polarization as accurately as did RNA microarray and LC/MS-MS. Then, we assessed the gene signature on dermal fibroblasts isolated from the experimental murine model. We observed a proinflammatory polarization at day 7, and a mixture of a proinflammatory and profibrotic polarizations at day 42 in line with histological findings. Our approach provides a small-size and convenient gene signature to assess murine fibroblast polarization.