ADOPTIVE IMMUNOTHERAPY FOR POSTTRANSPLANT EBV LYMPHOMA

EBV特异性细胞毒性T细胞(CTL)已被证明是 有效预防和治疗EBV感染 干细胞移植患者的淋巴增生性疾病，以及 人类白细胞抗原相合供者淋巴细胞过继免疫治疗 也适用于器官移植患者 紊乱，导致疾病完全缓解。这个 目前的建议是管理人类白细胞抗原相合或半相合 EBV-LPD器官移植患者的EBV CTL及监测 对这些患者的临床和放射学反应。有耐心的 外周血样本将在输液后进行检查 小卫星聚合酶链式反应检测供者淋巴细胞 DNA供体CTL将在输液后每隔一段时间进行免疫表型鉴定。 EBV特异性细胞毒性T淋巴细胞前体频率将是 在输液后每隔一段时间获得。细胞毒性的本质 将根据患者电话和捐赠者EBV来评估应答 CTL对MHC的限制，抑制程度 针对第I类的单抗的细胞毒性， 和CD-3，以及B-7/CD28共刺激在人外周血中的作用 供者CTL的扩大和患者对EBV的CTL反应。《The T》 细胞受体(TCR)VB表型将在输液后表征 T细胞和患者淋巴细胞与疾病的相关性 状态、体外CTL反应和TCR谱系。EBV CTL将 也可以从EBV血清阴性的捐赠者和脐带血中培养 单个核细胞检测EB病毒BLCL的免疫应答 没有感染过这种病毒的个人。作为以下内容的一部分 这一目标，患者的淋巴细胞也将与 自体EBV BLCL以扩大EBV反应性CTL的种群。 从患者体内获得的多克隆EBV CTL也将被克隆 用亚促分裂剂量的CD3和CD28进行扩展，目标是 扩增足够数量的这样的细胞 有可能用于治疗性输液。半定量EBV 所有器官移植患者将接受为期一年的聚合酶链式反应。 移植后每月间隔一次，以确定 EBV-LPD的存在与血清IL-6水平升高的相关性 从外周血淋巴细胞中提取EB病毒DNA。这项测试可能会有用。 在这些患者中早期诊断EBV-LPD时， 它们的淋巴增殖更有可能是由 减少免疫抑制。

EBV specific cytotoxic T cells (CTL) have been shown to be effective in preventing and treating EBV induced lymphoproliferative disease in stem cell transplant patients, and adoptive immunotherapy with HLA identical donor lymphocytes has also been applied to an organ transplant patient with this disorder, resulting in complete remission of the disease. The present proposal is to administer HLA identical or haplo-identical EBV CTL to organ transplant patients with EBV-LPD and to monitor these patients for clinical and radiographic response. Patient peripheral blood specimens will be examined post infusion using minisatellite PCR analysis for the presence of donor lymphocyte DNA. Donor CTL will be immunophenotyped at intervals post-infusion. EBV specific cytotoxic T lymphocyte precursor frequencies will be obtained at intervals post infusion. The nature of the cytotoxic response will be evaluated from patient calls as well as donor EBV CTL with regard to MHC restriction, the degree of inhibition of cytotoxicity with monoclonal antibodies directed against Class I, HLA-DR, and CD-3, and the role of B-7/CD28 co-stimulation in the expansion of donor CTL and in patient CTL responses to EBV. The T cell receptor (TCR) VB phenotype will be characterized on infused T cells as well as on patient lymphocytes to correlate disease status, in vitro CTL response, and TCR repertoire. EBV CTL will also be cultivated from EBV sero-negative donors and cord blood mononuclear cells to examine the immune response to EBV BLCL in individuals without a prior infection with this virus. As part of this goal, patient lymphocytes will also be cultured with autologous EBV BLCL to expand populations of EBV reactive CTL. Cloned as well a polyclonal EBV CTL obtained from patients will be expanded with submitogenic doses of CD3 and CD28, with the goal of expanding sufficient number of these cells that could be potentially used for therapeutic infusion. Semi-quantitative EBV PCR will be performed on all organ transplant patients for one year post transplant at monthly intervals in order to determine a correlation between then presence of EBV-LPD and elevated levels of EBV DNA from peripheral blood lymphocytes. This test may be useful in early diagnosis of EBV-LPD in these patients, at a time when their lymphoproliferations are more likely to be controlled by decreased immunosuppression.