Project description:Genomewide microarray analysis of murine tolerant, self-antigen specific CD8 T cells to identify genes and pathways underlying peripheral T cell tolerance Gene signature of tolerant CD8 T cells was compared to the signatures of naïve T cells, memory T cells, rescued T cells (=tolerant T cells undergoing homeostatic proliferation in lymphopenic, tolerogenic Alb:GAG mice), and re-tolerized T cells (=previously rescued T cells post homeostatic proliferation isolated from lymphoreplete wild-type B6 mice). Total RNA obtained from various sort-purified transgenic CD8 T cell subsets (naïve, memory, tolerant, rescued, and re-tolerized) isolated from spleens of different host mice
Project description:Genomewide microarray analysis of murine tolerant, self-antigen specific CD8 T cells to identify genes and pathways underlying peripheral T cell tolerance Gene signature of tolerant CD8 T cells was compared to the signatures of naïve T cells, memory T cells, rescued T cells (=tolerant T cells undergoing homeostatic proliferation in lymphopenic, tolerogenic Alb:GAG mice), and re-tolerized T cells (=previously rescued T cells post homeostatic proliferation isolated from lymphoreplete wild-type B6 mice).
Project description:Role for naturally occurring CD4+CD25+Foxp3+ regulatory T cells (nTregs) in counterbalancing this process. Using a transgenic murine model for autoimmune-mediated lung disease, we demonstrated that, despite pulmonary inflammation, lung-specific CD8+ T cells can reside quiescently in close proximity to self-antigen. Whereas self-reactive CD8+ T cells in the inflamed lung and lung-draining lymph nodes down-regulated the expression of effector molecules, those located in the spleen appeared to be partly antigen-experienced and displayed a memory-like phenotype. Since ex vivo-reisolated self-reactive CD8+ T cells were very well capable to respond to the antigen in vitro, we investigated a possible contribution of nTregs to the immune control over autoaggressive CD8+ T cells in the lung. We isolated antigen-specifc CD8+ T-cells from lungs and bronchial lymphnodes derived from chronic diseased mice (SPC-HAxCL4), healthy control mice (CL4) and acute influenza infected control mice (CL4+IAV) and perormed mRNA expression profiling of isolated CD8+ T cells. Each group represents a pool of at least n=4 animals. CD8+ T cell type comparison; lung disease state analysis
Project description:Self-specific CD8+ T cells often escape clonal deletion, but the properties and capabilities of such cells in a physiological setting are unclear. We characterized polyclonal CD8+ T cells specific for the melanocyte antigen tyrosinase-related protein 2 (Trp2) in mice that express or lack this enzyme due to deficiency in Dct, which encodes Trp2. The size, phenotype, and gene expression profile of the pre-immune Trp2/Kb-specific pool were similar in wild-type (WT) and Dct-deficient (Dct-/-) mice. Despite comparable initial responses to Trp2 immunization, WT Trp2/Kb-specific cells showed blunted expansion, and scRNAseq revealed WT cells less readily differentiated into a CD25+ proliferative population. Functional self-tolerance clearly emerged when assessing immunopathology: adoptively transferred WT Trp2/Kb-specific cells mediated vitiligo much less efficiently. Hence, CD8+ T cell self-specificity is poorly predicted by precursor frequency, phenotype or even initial responsiveness, while deficient activation-induced CD25 expression and other gene expression characteristics may help to identify functionally tolerant cells.
Project description:To describe the protein profile in hippocampus, colon and ileum tissue’ changing after the old faeces transplants, we adopted a quantitative label free proteomics approach.