VACCINE THERAPY FOR MULTIPLE MYELOMA UTILIZING ALLOGENEIC DENDRITIC CELLS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。假设：1.非清髓性异基因骨髓移植后接种同种独特型致敏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期研究将主要评估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。 鉴定移植前血清IgG同种型单克隆帕拉蛋白且符合移植条件的多发性骨髓瘤患者进行疫苗接种。在完成标准化疗之前，收集外周血用于疫苗开发。免疫球蛋白将通过柱色谱法从患者血清中分离，浓缩，纯化的Id蛋白将在与KLH结合前进行无菌过滤。在树突状细胞脉冲之前，将在无菌条件下使用戊二醛作为交联剂进行60 mg纯化Id蛋白与KLH的偶联。 使用最近开发的从外周血分离人树突状细胞的方法，这些细胞将从非清髓性移植供体中取出。在获得采集移植标本的同意书时，同种异体供体将同意采集单采标本。然而，在受者成功完成串联移植之前，将不会获得用于树突状细胞制备的单采产品。 使用Aastrom复制细胞系统（ARS）装置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群体。在37 ℃的培养箱中用补充有细胞因子（浓度为280单位/mL的GM-CSF和725单位/mL的IL 4）的冷冻培养基完成6天的初级孵育。然后采用无菌技术将Id-KLH蛋白（60 mg Id与1 mg/mL KLH偶联）加载到ARS中。二次成熟孵育持续48小时，之后从处理器自动收获细胞。收获后，洗涤细胞以去除细胞因子和其他物质，然后在20% DMSO和自体血浆中以单次使用等份冻存，每瓶10 x 106个细胞。随机选择一份冻存细胞的一次性等分试样，检测细菌、真菌和支原体污染沿着及内毒素含量。 将用Id-KLH脉冲的树突状细胞以1 × 106个活细胞的最小浓度重悬于1 mL 0.9%盐水中，并通过皮内给药给予。每种疫苗的给药部位将系统性交替。第一次疫苗接种将在同种异体移植后9至18个月进行，随后的五次疫苗接种将按每月一次的计划进行。

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.