VACCINE THERAPY FOR MULTIPLE MYELOMA UTILIZING ALLOGENEIC DENDRITIC CELLS

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。假设：1.非清髓性同种异体移植治疗多发性骨髓瘤后接种独特型致敏的同种异体树突状细胞治疗多发性骨髓瘤是安全可行的。2.接种疫苗的患者将产生可测量的独特型特异性免疫反应。实验设计：这项I/II期研究将初步评估30例多发性骨髓瘤患者接种同种异体树突状细胞自体ID-KLH疫苗的可行性和安全性。移植前血清中有免疫球蛋白同型单抗的多发性骨髓瘤患者和符合移植条件的患者被识别为接种疫苗。在完成标准化疗之前，采集外周血液用于疫苗开发。用柱层析法从患者血清中分离免疫球蛋白，浓缩，纯化的ID-蛋白在与KLH结合之前进行无菌过滤。60毫克纯化的ID蛋白与KLH的偶联将在无菌条件下进行，在树突状细胞脉冲之前使用戊二醛作为交联剂。使用最近开发的从外周血中分离人类树突状细胞的方法，这些细胞将从非清髓性移植供者身上提取。在获得同意采集移植标本时，同种异体捐献者将同意采集标本。然而，在受者成功完成串联移植之前，不会获得用于制备树突状细胞的分离产品。使用Astrom Replicell System(ARS)设备培养树突状细胞的自动化生产过程将扩大患者的树突状细胞数量。在37℃，添加细胞因子的冷冻液(GM-CSF浓度为280个单位/毫升，IL-4浓度为725个单位/毫升)中进行6天的原代培养。将ID-KLH蛋白(60 mg ID与KLH1 mg/mL偶联)通过无菌技术加载到ARS中。二次成熟孵化持续48小时，之后细胞自动从处理器中获取。收获后，将细胞洗去细胞因子和其他物质，然后在20%DMSO和自体血浆中冷冻保存在一次性等份中，每瓶10×106个细胞。随机选择一份一次性使用的冷冻保存细胞将进行细菌、真菌和支原体污染以及内毒素含量的测试。用ID-KLH冲击的树突状细胞将在1毫升0.9%生理盐水中以最低浓度为1×106活细胞重新悬浮，并通过皮内给药。给药地点将与每种疫苗系统地交替。第一次疫苗接种将在异基因移植后9至18个月之间进行，随后将按每月一次的计划接种五次疫苗。

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. Hypotheses: 1. Vaccination with allogeneic idiotype-pulsed dendritic cells following non-myeloablative allogeneic transplantation for multiple myeloma will be feasible and safe in treatment of this disease. 2. Vaccinated patients will mount a measurable idiotypic-specific immune response. Experimental Design: This Phase I/II study will evaluate primarily the feasibility and safety in thirty patients with multiple myeloma who will be vaccinated with allogeneic dendritic cell autologous Id-KLH vaccine. Patients with multiple myeloma with pre-transplant serum monoclonal para protein of the IgG isotype and who are eligible for transplantation are identified for vaccination. Prior to completing standard chemotherapy, peripheral blood is collected for vaccine development. Immunoglobulin will be isolated from patient serum by column chromatography, concentrated and the purified Id-protein will be sterile filtered prior to conjugation with KLH. The conjugation of the 60 mg purified Id-protein to KLH will be performed under aseptic conditions using glutaraldyde as the cross-linking agent prior to dendritic cell pulsing. Using recently developed methods for the isolation of human dendritic cells from peripheral blood, these cells will be taken from the non-myeloablative transplant donor. The allodonor will consent for apheresis specimen collection at the time that consent is obtained for transplant specimen collection. The apheresis product for dendritic cell preparation, however, will not be obtained until the recipient has successfully completed tandem transplant. An automated production process using the Aastrom Replicell System (ARS) device to grow dendritic cells will expand the patient's dendritic cell population. Primary incubation for 6 days is accomplished in the incubator at 37 0C with refrigerated medium supplemented by cytokines (GM-CSF at a concentration of 280 Units/mL and IL4 at 725 Units/mL). The Id-KLH protein (60 mg of Id conjugated to KLH at 1 mg/mL) will then be loaded into the ARS with aseptic technique. The secondary maturation incubation lasts for 48 hrs after which the cells are automatically harvested from the processor. After harvest, the cells will be washed to remove cytokines and other materials then cryopreserved in 20% DMSO and autologous plasma in single-use aliquots with 10 x 106 cells per vial. One single-use aliquot of cryopreserved cells selected at random will be tested for bacterial, fungal, and mycoplasma contamination along with endotoxin content. Dendritic cells pulsed with Id-KLH will be resuspended at a minimum concentration of 1 x106 viable cells in 1 mL of 0.9% saline and given by intradermal administration. Sites of administration will be systematically alternated with each vaccine. The first vaccination will occur between 9 and 18 months following allogeneic transplant and five subsequent vaccinations will be given on a once-monthly schedule.