DONOR BONE MARROW AND TRANSPLANT T AND B CELL REGULATION

描述（由申请人提供）：1994年9月，随后在 1996年11月，该中心开始了供者特异性骨髓的研究， 在尸体肾移植受者中首次输注DBMC细胞（n=63）， 然后是活体亲属供肾受者（n=48）。目标是（和 仍然是）通过识别状态最终撤销免疫抑制 DBMC可能诱导的操作性免疫耐受， 持续监控。出现了两个重要的观察结果：1）有 DBMC嵌合体与独特的免疫调节作用正相关; 2)DBMC具有明确的长期（5年）临床获益 DBMC嵌合体在移植肾存活率方面的研究方案， 骨髓腔在受体嵌合骨髓中，（受体- 来源的）供体细胞（RdD）和（受体来源的）受体细胞（RdR）可以 被隔离和测试。同种免疫抑制因子依次增加， 术后2年内RdD的影响，具有表型特征 这些长期嵌合细胞的。DBMC已培养3个月， 1年的受体（同种异体）饲养细胞（DBMC-L），后者是一个在 体外模型有点类似于体内RdD细胞生成。我们的具体 本提案的目标是：A.为了确定 调节（免疫调节）DBMC、RdD、DBMC-L和RdR细胞， 间接抗原呈递途径。B。来分析那些 细胞以诱导特异性T抑制细胞，即，感染耐受性C.到 评估这些细胞对应答细胞活化途径的影响， 进行TH 1-TH 2细胞因子的分子分析。D.分析总计 细胞mRNA表达与来自相同组织的类似处理的脾细胞相比， 通过差异表达和cDNA阵列来描述其在尸体供体中的表达。 独特的免疫调节特性。E.继续跟进受助人 用于供体嵌合CD 34+、CD 3、CD 19+和其他骨髓和淋巴细胞的水平 谱系细胞通过PCR流和细胞分离程序，从而关联 这些研究与具体目标二。二.继续开发可复制的 监测免疫抑制剂戒断的多方面方案（现在， 通过方案活检、有限稀释分析、同种抗体和 细胞因子ELISPOT研究和rag-i敲除小鼠中的体内试验。

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.