Immune responses to T-cell expansion + PCV immunization

描述（由申请人提供）：高剂量化疗后自体干细胞移植被认为是多发性骨髓瘤最有效的治疗方法，但仅约三分之一的患者完全缓解，治愈率极低。我们临床研究的长期目标是了解如何在自体干细胞移植的背景下重建抗骨髓瘤T细胞反应，以改善该程序的结果。这项工作背后的一个关键假设是，使用抗CD 3/抗CD 28缀合的磁珠离体共刺激自体T细胞可能有助于恢复T细胞对骨髓瘤肿瘤靶点的识别和反应。我们已经开发并实施了一项临床试验，其中骨髓瘤患者在自体移植后接受离体共刺激的自体T细胞输注。在该试验中，还用T细胞依赖性肺炎球菌缀合物疫苗（PCV）免疫患者，以提供免疫功能的可靠“读数”并测试施用离体扩增的T细胞是否会增强移植后的细胞或体液应答。使用随机设计，我们计划分析疫苗和“疫苗致敏”共刺激T细胞对整体肺炎球菌免疫应答的具体贡献。在本申请中，我们特别计划i）分析输注共刺激T细胞后免疫细胞重建的表型模式和机制; ii）测量针对结合疫苗和PCV（CRM-197）蛋白载体组分的细胞免疫应答; iii）测量针对VZV、CMV和EBV抗原的T细胞应答。将外周血单核细胞与PCV疫苗、CRM-197载体蛋白或CMV/VZV/EBV病毒抗原一起孵育，并通过增殖或ELISPOT试验检测分泌干扰素γ的T细胞应答者。从这些研究中获得的知识将提供一个平台，在此基础上开发多发性骨髓瘤和其他免疫应答血液肿瘤的联合免疫治疗策略，以及免疫功能低下宿主的免疫新方法。

DESCRIPTION (provided by applicant): High-dose chemotherapy followed by autologous stem cell transplantation is considered to be the most effective therapy for multiple myeloma but produces complete remissions in only about a third of patients and very few cures. The long-term objective of our clinical research is to learn how to re-establish anti-myeloma T-cell responses in the setting of autologous stem cell transplantation in order to improve upon the outcomes from this procedure. A key hypothesis behind this work is that ex-vivo costimulation of autologous T-cells using anti-CD3/anti-CD28 - conjugated magnetic beads may help to restore T-cell recognition and responsiveness toward myeloma tumor targets. We have developed and implemented a clinical trial in which patients with myeloma receive infusions of ex-vivo costimulated autologous T-cells after autotransplantation. In this trial, patients are also immunized with the T-cell dependent pneumococcal conjugate vaccine (PCV) in order to provide a reliable "readout" of immune function and to test whether administration of the ex-vivo expanded T-cells will enhance cellular or humoral responses post-transplantation. Using a randomized design, we plan to analyze the specific contributions of the vaccine and the "vaccine-primed" costimulated T-cells to the overall pneumococcal immune responses. Under this application, we specifically plan to i) analyze the phenotypic pattern and mechanism of immune cell reconstitution after infusion of costimulated T-cells; ii) measure the cellular immune responses that are directed against the conjugate vaccine and the protein carrier component of the PCV (CRM-197); iii) measure T-cell responses against VZV, CMV, and EBV antigens. Peripheral blood mononuclear cells will be incubated with PCV-vaccine, CRM-197 carrier protein, or CMV/VZV/EBV viral antigens and interferon gamma secreting T-cell responders will be detected by proliferation or ELISPOT assays. Knowledge gained from these studies will provide a platform on which to base the development of combined immunotherapy strategies for multiple myeloma and other immunoresponsive hematologic neoplasms as well as novel approaches to the immunization of immunocompromised hosts.