DONOR BONE MARROW AND TRANSPLANT T AND B CELL REGULATION

简介(申请人提供)：1994年9月，其后 1996年11月，该中心开始研究捐献者特有的骨髓。 身体肾移植受者首次输注细胞(DBMC)63例 然后是活体亲属供肾的受者(n=48)。目标是(和 继续)通过识别状态最终取消免疫抑制 可能由DBMC诱导的可操作性免疫耐受 持续监控。出现了两个重要的观察结果：1) DBMC嵌合体与独特的免疫调节作用呈正相关； 2)DBMC具有明确的长期(5年)临床益处 DBMC嵌合体扩大的同种移植肾存活方案 骨髓室。在受者的嵌合骨髓中，(受者- 派生)供体细胞(RDD)和(受体派生)受体细胞(RDR)可以 被隔离和检测。有顺序增加的同种免疫抑制 RDD术后最长2年的疗效，并有表型特征 这些长期嵌合细胞。DBMC已经培养了3个月到 接受(异基因)饲养层细胞1年(DBMC-L)，后者在 体外模型有点类似于体内RDD细胞的生成。我们的特定 本提案的目的是：内务部确定 调节(免疫调节)DBMC、RDD、DBMC-L和RDR细胞对直接和 间接抗原提呈途径。B.分析这些机构的能力 细胞诱导特定的T抑制细胞，即感染耐受。C.至 评估这些细胞对反应细胞激活途径的影响， 进行TH1-TH2细胞因子的分子分析。D.分析合计 细胞mRNA的表达与相同处理的脾细胞的比较 利用差异表达和基因芯片技术研究身体供体 独特的免疫调节特性。E.继续跟进受助人 供者嵌合CD34+、CD3、CD19+和其他髓系和淋巴系的水平 通过聚合酶链式反应和细胞分离程序对细胞进行谱系鉴定，从而将 这些研究具有特定的目标二.二.继续开发可重复使用的 监测免疫抑制停药的多方面方案(现已在 进展)通过常规活检、有限稀释分析、同种异体抗体和 细胞因子ELISPOT研究，以及RAG-I基因敲除小鼠的体内试验。

DESCRIPTION (provided by applicant): In September of 1994, and subsequently in November of 1996, this center embarked on studies of donor-specific bone marrow cell (DBMC) infusion first in cadaver renal transplant recipients (n=63) and then in recipients of living-related donor kidneys (n=48). The goal was (and continues to be) to eventually withdraw immunosuppression by identifying states of operational immunological tolerance that might be induced by DBMC and continuously monitored. Two significant observations have emerged: 1) There is a positive correlation of DBMC chimerism with unique immunoregulatory effects; 2) there is a definitive long-term (5 year) clinical benefit of the DBMC protocol on kidney allograft survival in which DBMC chimerism has expanded in the bone marrow compartment. In the recipient chimeric marrow, (recipient- derived) donor cells (RdD) and (recipient- derived) recipient cells (RdR) can be isolated and tested. There are sequentially increasing alloimmune inhibitory effects of RdD up to 2 years postoperatively, with phenotypic characterization of these long-term chimeric cells. DBMC has been cultured between 3 months and 1 year with recipient (allogeneic) feeder cells (DBMC-L), the latter an in vitro model somewhat analogous to RdD cell generation in vivo. Our specific aims in the present proposal are: I. A. To determine the influence of the modulating (immunoregulatory) DBMC, RdD, DBMC-L, and RdR cells on direct and indirect antigen presentation pathways. B. To analyze the capacity of those cells to induce specific T suppressor cells, i.e., infectious tolerance. C. To evaluate the effects of these cells on responding cell activation pathways, performing molecular analyses of TH1-TH2 cytokines. D. To analyze total cellular mRNA expression versus similarly treated spleen cells from the same cadaver donor by differential expression and cDNA array to delineate their unique immunoregulatory properties. E. To continue to follow up the recipients for levels of donor chimeric CD34+, CD3, CD19+ and other myeloid and lymphoid lineage cells by PCR Flow and cell isolation procedures, thereby correlating these studies with Specific Aim II. II. To continue to develop a reproducible multi-faceted protocol to monitor immunosuppressive withdrawal (now in progress) by protocol biopsy, limiting dilution analyses, alloantibody and cytokine ELISPOT studies, and trans-vivo assays in the rag-i knock-out mouse.