TREATMENT OF HODGKIN DISEASE WITH EBV SPECIFIC CTK

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我们建议开发和测试一种创新的方法来过继免疫治疗霍奇金病（HD）。在免疫功能正常的个体中，大约50%的HD病例与恶性Reed-Sternberg细胞及其变体中的许多EB病毒（EBV）衍生抗原的表达相关。其中，LMP 1和2抗原代表了可验证的和独特的肿瘤抗原，并且可以用作细胞毒性T淋巴细胞（CTL）介导的免疫疗法的靶标。我们以前曾成功地使用过继性CTL转移来预防和治疗骨髓移植后EBV相关的免疫母细胞性淋巴瘤。通过在输注前对CTL进行遗传标记，我们能够在体内追踪它们并分析它们的安全性，功能和寿命。已经表明，在移植后淋巴细胞增殖（LPD）的EBV阳性细胞对免疫治疗敏感，我们假设，恶性细胞霍奇金病，表达更有限的库EBV编码的抗原，也是合适的免疫治疗的目标。在特异性目标1中，我们将使用自体EBV转化的B细胞系作为刺激细胞，从复发性霍奇金病患者中产生基因标记的EBV特异性CTL。通过追踪标记基因，我们将测试注入的CTL在体内存活和扩增、安全并且含有足够的LMP-1/2特异性效应细胞以产生抗肿瘤活性的假设。在具体目标2中，我们将测试以下假设：使用经修饰以表达适当刺激物抗原的自体树突状细胞，可以从患者制备CD 4+和CD 8 + LMP 1和LMP 2a特异性CTL系。 对LMP 1和/或LMP 2a具有明确特异性的CTL细胞系（单特异性CTL）可能比多特异性细胞系更有效，在特异性目标3中，我们将检验以下假设：来自复发性霍奇金病患者的基因标记的LMP 1和LMP 2a特异性CTL细胞系在体内存活并持续存在，是安全的，增加患者对LMP 1和LMP 2a的免疫应答，并具有抗肿瘤作用。这些研究将提供适用于各种病毒和非病毒来源的人类恶性肿瘤的类似治疗的信息。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We propose to develop and test an innovative approach to adoptive immunotherapy for Hodgkin disease (HD). Approximately 50% of cases of HD in immunocompetent individuals are associated with expression of a number of Epstein-Barr virus (EBV) derived antigens in the malignant Reed-Sternberg cells and their variants. Of these, the LMP1 and 2 antigens, represent a verifiable and unique tumor antigen and can be used as a target for cytotoxic T lymphocyte (CTL)-mediated immunotherapy. We have previously used the adoptive transfer of CTL successfully to prevent and treat EBV-related immunoblastic lymphoma post bone marrow transplant. By genetically marking the CTL before infusion, we were able to track them in vivo and analyze their safety, function and longevity. Having shown that the EBV-positive cells in post-transplant lymphoproliferation (LPD) are susceptible to immunotherapy, we hypothesize that the malignant cells in Hodgkin disease, which express a more restricted repertoire of EBV encoded antigens, are also suitable targets for immunotherapy. In Specific Aim 1 we will generate gene-marked EBV-specific CTL from patients with relapsed Hodgkin disease, using autologous EBV-transformed B cells lines as stimulator cells. By tracking the marker gene, we will test the hypotheses that infused CTL survive and expand in vivo, are safe and contain sufficient LMP-1/2 specific effector cells to generate anti-tumor activity. In specific Aim 2, we will test the hypotheses that CD4+ and CD8+ LMP1 and LMP2a-specific CTL lines can be prepared from patients, using autologous dendritic cells modified to express the appropriate stimulator antigens. CTL lines with defined specificity against LMP1 and or LMP2a (monospecific CTL) may be more effective than polyspecific lines, and in Specific Aim 3, we will test the hypotheses that gene-marked LMP1- and LMP2a-specific CTL lines from patients with relapsed Hodgkin disease, survive and persist in vivo, are safe, increase patient immune responses to LMP1 and LMP2a and have anti-tumor effects. The studies will provide information applicable to similar therapy for a variety of human malignancies of viral and non-viral origin.