IMMUNOTHERAPY WITH CMV AND EBV SPECIFIC T LYMPHOCYTES

描述：（申请人的摘要）人巨细胞病毒（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CMV的治疗依赖于引起 骨髓抑制和肾毒性，从而限制了它们的应用。里德尔和 同事已经证明SCT患者可以达到正常的CMV水平， 输注供体来源的CMV特异性 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），如供体来源的皮肤 成纤维细胞，以及T细胞克隆。申请人的实验室已编码 pp 65是逆转录病毒（MSCV）中的免疫显性CMV肽， 成功转导的B淋巴母细胞样细胞系（BLCL）和SF， 表达这种肽。使用单个APC，他开发了CMV pp 65， EBV特异性多克隆CTL。这些CTL是MHC I类限制性的和CD 8+。 本研究旨在探讨CMV/EBV特异性CTL细胞培养的可行性 使用来自干细胞供体的pp 65转导的BLCL。他将应用这些 干细胞移植患者预防CMV和EBV的双特异性CTL 接受T细胞耗竭（TCD）、匹配或部分匹配的相关供体 移植他将描述这些人的CMV和EBV特异性免疫力， 患者在输注后间隔，并检查TCR V β库， 培养的CTL和患者PBMC的HLA限制。由于风险 移植物抗宿主病（GVHD），如果使用非特异性供体PBMC 免疫疗法，以及抗CMV药物的发病率，这种形式的治疗 可能会对这一患者群体产生重大影响。使用定量， CMV的竞争性PCR测定，他将测量来自 CMV/EBV CTL输注的接受者的外周血。半定量 PCR检测将用于测量EBV DNA水平。收养时 免疫疗法对于接受外周血干细胞移植的患者是可能的 细胞或骨髓移植，这不是一个选择，为接受者的脐带 血液移植或有CMV血清阴性供体的患者。作为 作为这些研究的一部分，申请人将研究以下可能性： 使用来自CMV血清阴性供体的pp 65 BLCL开发CMV特异性CTL， 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.