Project description:To illustrate the functional heterogeneity of T cells in the heart allograft rejection,we established the murine heterotopic heart transplantation model and isolated CD45 positive cells from cardiac grafts and spleens for single cell transcriptome and TCR sequencing.
Project description:T cells primarily drive the autoimmune destruction of pancreatic beta cells in Type 1 diabetes (T1D). However, the profound yet uncharacterized diversity of the T cell populations in vivo has hindered obtaining a clear picture of the T cell changes that occur longitudinally as T1D onset happens. Here we profiled the transcriptome and TCR repertoire of T cells at single-cell resolution from longitudinally collected peripheral blood and islets of Non-Obese Diabetic (NOD) mice using single-cell RNA sequencing technology. We detected clonal expansion and characterized the transcriptional landscape of "islets-matching" T cell clones in the blood and "blood-matching" T cell clones in islets of diabetic NOD mice using the TCR as a molecular barcode. The clonally matching cells show enriched interferon-gamma response pathways than non-matching cells in blood and islets. In addition, we identified a set of transcriptional markers associated with the matching status of the T cell clones. This study provides a single-cell level transcriptome and TCR repertoire atlas of T cells in NOD mice. The results show that TCR and transcriptional signature can be used combinedly to develop biomarker panels for tracking and investigating islets infiltrating T cells.
Project description:Single-cell TCR sequencing of T cells from ANCA-associated glomerulonephritis patients and experimental crescentic glomerulonephritis mouse model.
Project description:We have reported that intra-tumoral treatment with 1V270, a phospholipid-conjugated TLR7 agonist,induces local expansion an systemic dispersion of oligoclonal tumor-specific T cells by TCR repertoire analysis using next generation RNAseq methodology. Here, we examined whether systemic 1V270 therapy also induced oligoclonal expansion of tumor-specific T cells. Two groups of BALB/c mice (n=4/group) were i.p. treated with 1V270,a phospholipid-conjugated TLR7 agonist. One cohort of mice was i.v. injected with 4T1-GLF cells (2×104) on day 0. Another cohort did not receive i.v. tumor injection (no tumor-exposed mice). 4T1 cells were orthotopically inoculated on day 21. To examine clonal specificity of tumor-specific T cells, CD8+ cells were isolated from the spleens and the tumor infiltrating lymphocytes of secondarily challenged tumors after initial 1V270 therapy. The TCR repertoires were assessed by next generation RNA sequencing of both TCRαand TCR β genes.
Project description:Single-cell RNA-sequencing and single-cell TCR-sequencing was used to characterize PGE2-mediated changes in the gene expression profile of tumor-infiltrating CD8+ T cells.
Project description:ACh was originally isolated from spleen back in 1929, however, its contribution to immune regulation has only recently been appreciated. Subsets of both T and B lymphocytes have been found to express choline acetyltransferase (Chat) and produce ACh. To date, Chat-expressing T cells have been described as relaying neural signals in the spleen to modulate immune responses; regulating blood pressure; and promoting anti-viral immune responses. In a murine multi-hit model of hepatocellular carcinoma, we observed activation of the adaptive immune response and induction of Chat-expressing CD4+ T cells. These cholinergic T cells curtail the development of liver cancer by supporting anti-tumor immune responses. We used single-cell RNA sequencing to investigate the transcriptome and TCR repertoire of the Chat-expressing CD4+ T cells in healthy and HCC-bearing livers.
Project description:Thymic regulatory T cells (tTreg) are critical in maintenance of normal T cell immunity and tolerance. The role of TCR in tTreg cell selection remains incompletely understood. Here we assessed TCRa and TCRb sequences of tTreg and conventional thymic CD4+ T (Tconv) cells by high throughput sequencing. We identified ab TCR sequences that were unique to either tTreg or Tconv cells and found that these were distinct as recognized by machine learning (ML) algorithm and by preferentially used amino acid trimers in ab CDR3 of tTreg cells. In addition, a proportion of ab TCR sequences expressed by tTreg were also found in Tconv cells, and ML classified the great majority of these shared ab TCR sequences as characteristic of Tconv and not tTreg cells. These findings identify two populations of tTreg, one in which Treg fate is associated with unique properties of the TCR, and another with TCR properties characteristic of Tconv cells for which tTreg fate is determined by factors beyond TCR sequence.