The use of autoantigen-specific regulatory T cells (Tregs) as a cellular therapy for autoimmune diseases is appealing. However, it is challenging to isolate and expand large quantity of Tregs expressing disease-relevant T-cell receptors (TCR). To overcome this problem, we used an approach aiming at redirecting the specificity of polyclonal Tregs through autoreactive TCR gene transfer technology. In this study, we examined whether Tregs engineered through retroviral transduction to express a TCR cross-reactive to two CNS autoantigens, myelin oligodendrocyte glycoprotein (MOG) and neurofilament-medium (NF-M), had a superior protective efficacy compared with Tregs expressing a MOG mono-specific TCR. We observed that engineered Tregs (engTregs) exhibited in vitro regulatory effects related to the antigenic specificity of the introduced TCR, and commensurate in potency with the avidity of the transduced TCR. In experimental autoimmune encephalomyelitis (EAE), adoptively transferred engTregs proliferated, and migrated to the CNS, while retaining FoxP3 expression. EngTregs expressing MOG/NF-M cross-reactive TCR had superior protective properties over engTregs expressing MOG-specific TCR in MOG-induced EAE. Remarkably, MOG/NF-M bi-specific TCR-engTregs also improved recovery from EAE induced by an unrelated CNS autoantigen, proteolipid protein (PLP). This study underlines the benefit of using TCRs cross-reacting towards multiple autoantigens, compared with mono-reactive TCR, for the generation of engTregs affording protection from autoimmune disease in adoptive cell therapy.

使用自身抗原特异性调节性T细胞（Tregs）作为自身免疫性疾病的细胞疗法很有吸引力。但是，分离和扩增大量表达疾病相关T细胞受体（TCR）的Treg具有挑战性。为了克服这个问题，我们使用了一种旨在通过自身反应性TCR基因转移技术重定向多克隆Tregs特异性的方法。在这项研究中，我们研究了通过逆转录病毒转导工程改造的Tregs是否表达与两种CNS自身抗原，髓磷脂少突胶质细胞糖蛋白（MOG）和神经丝介质（NF-M）交叉反应的TCR，与表达Tregs的Tregs相比具有更好的保护功效。 MOG单特异性TCR。我们观察到工程化的Tregs（engTregs）在体外表现出与引入的TCR的抗原特异性有关的调节作用，以及与转导的TCR的亲和力相当的效力。在实验性自身免疫性脑脊髓炎（EAE）中，过继转移的engTregs增殖并迁移到CNS，同时保留FoxP3表达。在MOG诱导的EAE中，表达MOG / NF-M交叉反应性TCR的EngTreg具有优于表达MOG特异性TCR的engTregs的保护特性。值得注意的是，MOG / NF-M双特异性TCR-engTregs还改善了由无关CNS自身抗原，蛋白脂蛋白（PLP）诱导的EAE恢复。这项研究强调了与单反应性TCR相比，使用与多种自身抗原交叉反应的TCR产生engTreg的益处，该engTreg在过继细胞治疗中提供了针对自身免疫性疾病的保护。