Spanier Justin A, Fung Vivian, Wardell Christine M, Alkhatib Mohannad H, Chen Yixin, Swanson Linnea A, Dwyer Alexander J, Weno Matthew E, Silva Nubia, Mitchell Jason S, Orban Paul C, Mojibian Majid, Verchere C Bruce, Fife Brian T, Levings Megan K
Center for Immunology, University of Minnesota Medical School, Minneapolis, MN, USA.
Center for Autoimmune Disease Research, Department of Medicine, University of Minnesota Medical School, Minneapolis, MN, USA.
bioRxiv. 2023 Feb 24:2023.02.23.529737. doi: 10.1101/2023.02.23.529737.
Adoptive immunotherapy with Tregs is a promising approach for prevention or treatment of type 1 diabetes. Islet antigen-specific Tregs have more potent therapeutic effects than polyclonal cells, but their low frequency is a barrier for clinical application. To generate Tregs that recognize islet antigens, we engineered a chimeric antigen receptor (CAR) derived from a monoclonal antibody with specificity for the insulin B-chain 10-23 peptide presented in the context of the IA MHC class II allele present in NOD mice. Peptide specificity of the resulting InsB-g7 CAR was confirmed by tetramer staining and T cell proliferation in response to recombinant or islet-derived peptide. The InsB-g7 CAR re-directed NOD Treg specificity such that insulin B 10-23-peptide stimulation enhanced suppressive function, measured via reduction of proliferation and IL-2 production by BDC2.5 T cells and CD80 and CD86 expression on dendritic cells. Co-transfer of InsB-g7 CAR Tregs prevented adoptive transfer diabetes by BDC2.5 T cells in immunodeficient NOD mice. In wild type NOD mice, InsB-g7 CAR Tregs stably expressed Foxp3 and prevented spontaneous diabetes. These results show that engineering Treg specificity for islet antigens using a T cell receptor-like CAR is a promising new therapeutic approach for the prevention of autoimmune diabetes.
Chimeric antigen receptor Tregs specific for an insulin B-chain peptide presented by MHC class II prevent autoimmune diabetes.
用调节性T细胞（Tregs）进行过继性免疫疗法是预防或治疗1型糖尿病的一种有前景的方法。胰岛抗原特异性Tregs比多克隆细胞具有更强的治疗效果，但其低频率是临床应用的一个障碍。为了产生识别胰岛抗原的Tregs，我们构建了一种嵌合抗原受体（CAR），该受体源自一种单克隆抗体，对在非肥胖糖尿病（NOD）小鼠中存在的IA MHC II类等位基因背景下呈递的胰岛素B链10 - 23肽具有特异性。通过四聚体染色以及对重组或胰岛来源肽的T细胞增殖反应，证实了所得InsB - g7 CAR的肽特异性。InsB - g7 CAR重新定向了NOD Treg的特异性，使得胰岛素B 10 - 23肽刺激增强了抑制功能，这通过BDC2.5 T细胞增殖和IL - 2产生的减少以及树突状细胞上CD80和CD86表达的降低来衡量。InsB - g7 CAR Tregs的共转移预防了免疫缺陷NOD小鼠中BDC2.5 T细胞的过继性转移糖尿病。在野生型NOD小鼠中，InsB - g7 CAR Tregs稳定表达Foxp3并预防了自发性糖尿病。这些结果表明，使用类似T细胞受体的CAR对胰岛抗原进行Treg特异性工程改造是预防自身免疫性糖尿病的一种有前景的新治疗方法。
对II类MHC呈递的胰岛素B链肽具有特异性的嵌合抗原受体Tregs预防自身免疫性糖尿病。
