Regulatory CD4/CD8 T cells in autoimmune demyelination

描述（由申请人提供）：免疫调节在控制成熟的自我反应性、潜在致病性T细胞中起关键作用。不同的细胞成分及其在调控机制中的相互作用尚不清楚。在这里，我们将研究一种显性负反馈机制如何控制髓鞘碱性蛋白（MBP）反应性Vbeta8.2+ T细胞介导H-2u小鼠（多发性硬化症的原型）的实验性自身免疫性脑脊髓炎。这一调控主要是通过调节CD8和CD4 T细胞（Treg）分别识别I类和II类MHC背景下的TCR肽/MHC复合物。我们提出（见图1）CD4 Treg有助于CD8 Treg的募集/激活，最终诱导活化的Vbeta8.2+ Th1细胞凋亡。致病性Th1的消耗和非致病性Th2细胞的扩增最终导致抗mbp反应的免疫偏差和对EAE的保护。CD4 Treg对框架3区肽BS有反应，而CD8 Treg识别来自TCR Vbeta8.2链的CDR1/2区肽B3或p41-50。通过CDR3长度谱分型、免疫镜分析和cfse标记细胞的膜联蛋白- v染色，我们将确定体内诱导或生理调节后Vbeta8.2+ T细胞的命运。我们将分别确定1型和2型细胞因子在调节的初始和后期阶段的作用。CD8 Treg克隆将生成并表征其mhc限制，TCR-V基因使用和功能特性，包括其体外和体内的细胞溶解活性，以及它们在过继转移时控制疾病的能力。我们将利用B10.PL确定CD8 Treg的杀伤机制。lpr B10.PL。黄金和黄金。穿孔素- / -小鼠。这里涉及的免疫原理的知识不仅对理解免疫调节的生物学很重要，而且对设计基于tcr的人类自身免疫性疾病治疗方法也很重要。

DESCRIPTION (provided by applicant): Immune regulation plays a critical role in the control of mature self-reactive, potentially pathogenic T cells. Different cellular components and their interactions in regulatory mechanisms are not well understood. Here we will investigate how a dominant negative feedback mechanism controls myelin basic protein (MBP)-reactive Vbeta8.2+ T cells mediating experimental autoimmune encephalomyelitis in the H-2u mouse, a prototype for multiple sclerosis. This regulation is focused on the recognition of TCR peptide/MHC complexes in class I and II MHC contexts by regulatory CD8 and CD4 T cells (Treg) respectively. We have proposed (see Figure 1) that CD4 Treg provide help in the recruitment/activation of CD8 Treg, which ultimately induce apoptosis in activated Vbeta8.2+ Th1 cells. The depletion of pathogenic Th1 and expansion of non-pathogenic Th2 cells eventually leads to immune deviation of the anti-MBP response and protection from EAE. CD4 Treg are reactive to a framework 3-region peptide, BS, whereas CD8 Treg recognize the CDR1/2 region peptide, B3 or p41-50, from the TCR Vbeta8.2 chain. Using CDR3 length spectratyping, or immunoscope analysis and annexin-V staining of CFSE-labeled cells we will determine the fate of Vbeta8.2+ T cells following induced or physiological regulation in vivo. We will determine the role of type 1 and type 2 cytokines in the initial and late phases of regulation, respectively. CD8 Treg clones will be generated and characterized for their MHC-restriction, TCR-V gene usage and functional properties, including their cytolytic activity both in vitro and in vivo, and their ability to control disease upon adoptive transfer. We will determine the mechanism of killing by the CD8 Treg using B10.PL.lpr, B10.PL.gld and B10.PL.perforin-/- mice. Knowledge of the immune principles involved here is important not only for understanding the biology of immune regulation but also in the design of TCR-based therapeutic approaches for autoimmune conditions in humans.