Immunoregulation by TLR-activated TIM-1+ ProB Cells in Transplantation

摘要(项目2) 与其使用被动转移分化良好的成人免疫调节细胞作为一种手段来创造 移植耐受，我们发现一群CpG激活的Pro-B细胞非常有效， 在每个细胞的基础上比Tregs强大得多，而且更容易制备。只转移了40,000，但不是 90,000，诱导MHC血型不合的胰岛和同种异体心脏移植的永久植入和可能的耐受 老鼠模型。我们已经发现CpG诱导了TIM-1的表达，TIM-1是一种潜在的主开关 免疫调节模块在成熟Bregs中的表达(见项目1/Kuchroo)。我们还了解到， 调控模型成员发现，抗TIM-1单抗连接TIM-1可诱导IL-10的表达 被库奇鲁。CpG的逐步贡献和TIM-1通路的激活通过 基于RNAseq和纳米串的分析。为了更准确地描述命运和表型，增殖性 对于CpG激活的Pro-B细胞的分化，始终使用谱系追踪的方法。我们知道 CpG激活的Pro-B细胞在转移后增殖和分化，当引入最佳，而不是 太多了，手机号码。根据环境信号，CpG激活的Pro-B细胞可以分化为 来自RAG KO小鼠的CpG激活的PRO-B细胞不产生Bregs。在这个项目中，我们将 分析耐受的细胞学基础认为CpG激活的B调节子及其后代创造了 有利于激活、扩大和保留供体特异性Tregs的环境。具体的但是 B细胞分化的多方面要求，包括利基损害扩增的性质和 区分CpG-Probe，在获得耐受性方面将通过使用面板仔细分析 选定的基因敲除小鼠。T和B调节子在诱导和维持耐受性中的作用将是 通过有选择地摧毁Tregs或CpG Activate成熟后代的时间进程协议确定 支持细胞。CpG-ProB子代的多样性及其IL-10调控模块的表达 将在移植时由CpG-pros及其后代的单细胞RNAseq进行检测。

Abstract (Project 2) Rather than use passive transfer of well differentiated adult immunoregulatory cells as a means to create transplant tolerance, we have discovered that a population CpG activated pro-B cells are extraordinarily potent, far more potent on a per cell basis than Tregs and far easier to prepare. Transfer of only 40,000, but not 90,000, induces permanent engraftment and probably tolerance in MHC incompatible islet and cardiac allograft mouse models. We have discovered that CpG induces expression TIM-1 a potential master switch for expression of an immunoregulatory module in mature Bregs (see Project 1/ Kuchroo). We also learned that ligation of TIM-1 by anti-TIM-1 mAb induces expression of IL-10, a member of the regulatory model discovered by Kuchroo. The stepwise contributions of CpG and activation of the TIM-1 pathway are examined through RNAseq and NanoString based analysis. To more precisely characterize the fate and phenotype, proliferative and differentiation of CpG activated pro-B cells, lineage-tracking methods are utilized throughout. We know that CpG activated pro-B cells proliferate and differentiate after transfer when introduced in optimal, not excessive, cell number. Depending upon environmental cues CpG activated pro-B cells can differentiate into a variety Bregs but CpG activated pro-B cells from RAG KO mice do not generate Bregs. In this project we will analyze the cellular basis of tolerance believing that CpG activated B regs and their progeny create an environment conducive to activation, expansion and retention of donor specific Tregs. The specific but multifaceted requirements for B cell differentiation, including the nature of the niche impairing expansion and differentiation of CpG-ProBs, in the acquisition of tolerance will be analyzed through use of a panel of carefully selected gene knockout mice. The role of T and B regs in the induction and maintenance of tolerance will be determined through time course protocols that selectively destroy Tregs or the mature progeny of CpG activate pro cells. The diversity of CpG-ProB progeny and their expression of a regulatory module that is IL-10 inclusive will be examined by single cell RNAseq of CpG-ProBs and their progeny at the transplant.