IMMUNOTHERAPY WITH CMV AND EBV SPECIFIC T LYMPHOCYTES

描述：(申请人摘要)人类巨细胞病毒(CMV)仍然是一种 异基因干细胞移植受者发病和死亡的严重原因 移植(SCT)。目前巨细胞病毒的治疗依赖于导致 骨髓抑制和肾毒性，因此限制了它们的使用。里德尔和 同事们已经证明，SCT患者可以达到正常的CMV水平 输注供体来源的CMV特异性细胞免疫 CTL.然而，这些技术受到所需时间和试剂的限制 培养抗原提呈细胞(APC)，如供体皮肤 成纤维细胞，以及T细胞克隆。申请人的实验室已经编码 Pp65，CMV的免疫优势肽，存在于逆转录病毒(MSCV)中，并具有 成功转导B淋巴母细胞系(BLCL)和SF实现 这种多肽的表达。使用单个APC，他已经开发出CMV pp65和 EBV特异性多克隆CTL。这些CTL是MHC I类限制性CTL和CD8+CTL。 这些研究将考察培养CMV/EBV特异性CTL的可行性 用pp65转导的干细胞供者的BLCL。他将应用这些 双特异性CTL预防干细胞移植患者巨细胞病毒和EB病毒感染 接受T细胞耗尽(TCD)、相合或部分相合的亲属供者 移植。他将描述这些病毒的CMV和EBV特异性免疫的特征 患者在输液后每隔一段时间检查TCR Vbeta谱系和 培养的CTL和患者PBMC的HL A限制。由于存在以下风险 移植物抗宿主病(GVHD)如果使用非特异性供者PBMC治疗 免疫治疗，以及抗CMV药物的发病率，这种治疗形式 可能会对这一患者群体产生重大影响。使用定量的， 竞争性聚合酶链式反应检测巨细胞病毒，他将从 巨细胞病毒/EB病毒CTL输注受者外周血。半定量的 采用聚合酶链式反应检测EB病毒DNA水平。在采用的同时 免疫治疗是接受外周血干细胞治疗的一种可能性 细胞或骨髓移植，这不是脐带血接受者的选择 血液移植或献血者CMV血清阴性的患者。AS 作为这些研究的一部分，申请者将研究 利用CMV血清阴性供体pp65 BLCL制备CMV特异性CTL 来自CB淋巴细胞。

DESCRIPTION: (Applicant's Abstract) Human cytomegalovirus (CMV) remains a serious cause of morbidity and mortality in recipients of allogeneic stem cell transplants (SCT). Current therapies for CMV rely upon drugs that cause myelosuppression and nephrotoxicity, thus limiting their use. Riddell and associates have demonstrated that SCT patients can achieve normal levels of CMV specific cellular immunity following infusions of donor-derived, CMV specific CTL. These technologies, however, are limited by the time and reagents required to cultivate antigen presenting cells (APC), such as donor-derived skin fibroblasts, as well as T cell clones. The applicant's laboratory has encoded pp65, the immunodominant CMV peptide, in a retrovirus (MSCV), and has successfully transduced B lymphoblastoid cells lines (BLCL) and SF to achieve expression of this peptide. Using a single APC, he has developed CMV pp65 and EBV specific, polyclonal CTL. These CTL are MHC Class I restricted and CD8+. These studies will examine the feasibility of cultivating CMV/EBV specific CTL using pp65 transduced BLCL from stem cell donors. He will apply these bi-specific CTL for CMV and EBV prophylaxis in stem cell transplant patients receiving T cell depleted (TCD), matched or partially matched related donor transplants. He will characterize the CMV and EBV specific immunity of these patients at intervals post-infusion and examine the TCR Vbeta repertoire and HLA restriction of cultivated CTL and patient PBMC. Due to risks of graft-versus-host disease (GVHD) if using non-specific donor PBMC for immunotherapy, as well as the morbidity of anti-CMV drugs, this form of therapy may have a significant impact on this patient population. Using a quantitative, competitive PCR assay for CMV, he will measure levels of CMV DAN from the peripheral blood of recipients of CMV/EBV CTL infusions. A semi-quantitative PCR assay will be used to measure levels of EBV DNA. While adoptive immunotherapy is a possibility for patients receiving peripheral blood stem cell or bone marrow transplants, it is not an option for recipients of cord blood transplants or for patients who have donors who are CMV seronegative. As a part of these studies, the applicant will examine the possibility of developing CMV specific CTL using pp65 BLCL from CMV seronegative donors and from CB lymphocytes.