Project description:Regulatory T cells (Tregs) are promising cellular therapies to induce immune tolerance in organ transplantation and autoimmune disease. The success of chimeric antigen receptor (CAR) T-cell therapy for cancer has sparked interest in using CARs to generate antigen-specific Tregs. Here, we compared CAR with endogenous T cell receptor (TCR)/CD28 activation in human Tregs. Strikingly, CAR Tregs displayed increased cytotoxicity and diminished suppression of antigen-presenting cells and effector T (Teff) cells compared with TCR/CD28 activated Tregs. RNA sequencing revealed that CAR Tregs activate Teff cell gene programs. Indeed, CAR Tregs secreted high levels of inflammatory cytokines, with a subset of FOXP3+ CAR Tregs uniquely acquiring CD40L surface expression and producing IFNγ. Interestingly, decreasing CAR antigen affinity reduced Teff cell gene expression and inflammatory cytokine production by CAR Tregs. Our findings showcase the impact of engineered receptor activation on Treg biology and support tailoring CAR constructs to Tregs for maximal therapeutic efficacy.
Project description:Reprogramming autoreactive CD4⁺ effector T (Teff) cells into immunosuppressive regulatory T (Treg) cells represents a promising strategy for treating established autoimmune diseases. However, the stability and function of such reprogrammed Tregs under inflammatory conditions remain unclear. Here, we show that demethylation of core Treg identity genes in Teff cells yields lineage-stable Effector T cell Reprogrammed Tregs (ER-Tregs). A single adoptive transfer of ER-Tregs not only prevents autoimmune neuroinflammation in mice when given before disease onset but also arrests its progression when administered after onset. Compared to Foxp3‑overexpressing Teff cells, induced Tregs from naïve precursors, and endogenous Tregs, ER‑Tregs provide superior protection against autoimmune neuroinflammation. This enhanced efficacy stems from their inherited autoantigen specificity and selectively preserved effector‑cell transcriptional programs, which together bolster their fitness in inflammatory environments and enhance their suppressive capacity. Our results establish epigenetic reprogramming of autoreactive Teff cells as an effective approach to generate potent, stable Tregs for the treatment of refractory autoimmune conditions.
Project description:Recent findings suggest that undifferentiated, stem-like, antigen specific T cells serve as an important long-term reservoir for autoimmune CD8 T cell responses. However, it is still unclear whether CD4 T cells exhibit a similar differentiation trajectory culminating in terminal differentiation, acquisition of an exhausted phenotype, and loss of stemness and function. We analyzed islet infiltrating T cells by scRNAseq and flow cytometry and found that while CD4 T cells in autoimmune diabetes share many features of exhaustion with CD8 T cells, expression patterns of inhibitory receptors are distinct in autoimmune T cells compared to T cells in chronic LCMV infection.
Project description:Low-dose recombinant interleukin-2 (rIL-2) therapy holds significant promise for treating autoimmune and inflammatory disorders by selectively expanding the endogenous pool of CD4⁺Foxp3⁺ regulatory T cells (Tregs). However, clinical benefits are limited by the suboptimal pharmacokinetic profile of rIL-2, which fails to support sustained Treg activation. Here we investigated the efficacy of a long-lasting IL-2-based biologic mIL-2/CD25 to limit type 1 diabetes in female NOD mice when administered in a manner to cause periodic versus continual increases in Tregs. We find that the latter is highly effective in limiting autoimmunity and leads to substantial remodeling of the pancreatic islet immune landscape, which has important implications for advancing IL-2-based therapies for autoimmunity.
Project description:Recent findings suggest that undifferentiated, stem-like, antigen specific T cells serve as an important long-term reservoir for autoimmune CD8 T cell responses. However, it is still unclear whether CD4 T cells exhibit a similar differentiation trajectory culminating in terminal differentiation, acquisition of an exhausted phenotype, and loss of stemness and function. We analyzed islet infiltrating T cells in 8- and 16-week old NOD mice by scRNAseq and flow cytometry and found that while CD4 T cells in autoimmune diabetes share many features of exhaustion with CD8 T cells, expression patterns of inhibitory receptors are distinct in autoimmune T cells compared to T cells in chronic LCMV infection.
Project description:Low-dose recombinant interleukin-2 (rIL-2) therapy holds significant promise for treating autoimmune and inflammatory disorders by selectively expanding the endogenous pool of CD4⁺Foxp3⁺ regulatory T cells (Tregs). However, clinical benefits are limited by the suboptimal pharmacokinetic profile of rIL-2, which fails to support sustained Treg activation. Here we investigated the efficacy of a long-lasting IL-2-based biologic mIL-2/CD25 to limit type 1 diabetes in female NOD mice when administered in a manner to cause periodic versus continual increases in Tregs. We find that the latter is highly effective in limiting autoimmunity and leads to substantial remodeling of the pancreatic islet immune landscape, which has important implications for advancing IL-2-based therapies for autoimmunity.
Project description:Seropositivity for autoantibodies against islet autoantigens is associated with the development of autoimmune type 1 diabetes and B cell targeted therapies are effective in both mouse models and in patients who are affected by or at risk for autoimmune type 1 diabetes. The role of B cell receptor affinity in autoimmune type 1 diabetes is unclear. Here, we employed single cell RNA sequencing to define the relationship between B cell receptor affinity for insulin and B cell phenotype during disease development using immunoglobulin heavy chain (VH125) transgenic mouse model (VH125.NOD) in which insulin binding B cells lose self-tolerance, becoming activated during development of autoimmmun type 1 diabetes.