Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC, USA; Department of Surgery, Duke University Medical Center, Durham, NC, USA; Department of Immunology, Duke University Medical Center, Durham, NC, USA.
Duke Human Vaccine Institute, Duke University Medical Center, Durham, NC, USA.
J Immunol Methods. 2014 Jul;409:21-30. doi: 10.1016/j.jim.2014.05.013. Epub 2014 Jun 5.
The Center for HIV/AIDS Vaccine Immunology (CHAVI) consortium was established to determine the host and virus factors associated with HIV transmission, infection and containment of virus replication, with the goal of advancing the development of an HIV protective vaccine. Studies to meet this goal required the use of cryopreserved Peripheral Blood Mononuclear Cell (PBMC) specimens, and therefore it was imperative that a quality assurance (QA) oversight program be developed to monitor PBMC samples obtained from study participants at multiple international sites. Nine site-affiliated laboratories in Africa and the USA collected and processed PBMCs, and cryopreserved PBMC were shipped to CHAVI repositories in Africa and the USA for long-term storage. A three-stage program was designed, based on Good Clinical Laboratory Practices (GCLP), to monitor PBMC integrity at each step of this process. The first stage evaluated the integrity of fresh PBMCs for initial viability, overall yield, and processing time at the site-affiliated laboratories (Stage 1); for the second stage, the repositories determined post-thaw viability and cell recovery of cryopreserved PBMC, received from the site-affiliated laboratories (Stage 2); the third stage assessed the long-term specimen storage at each repository (Stage 3). Overall, the CHAVI PBMC QA oversight program results highlight the relative importance of each of these stages to the ultimate goal of preserving specimen integrity from peripheral blood collection to long-term repository storage.
人类免疫缺陷病毒/艾滋病疫苗免疫学中心(CHAVI)联盟的建立是为了确定与 HIV 传播、感染和病毒复制控制相关的宿主和病毒因素,目的是推进 HIV 保护性疫苗的开发。为了实现这一目标,研究需要使用冷冻保存的外周血单个核细胞(PBMC)标本,因此必须制定质量保证(QA)监督计划,以监测来自多个国际地点的研究参与者的 PBMC 样本。非洲和美国的 9 个基地附属实验室收集和处理 PBMC,并将冷冻的 PBMC 运往非洲和美国的 CHAVI 存储库进行长期储存。根据良好临床实验室实践(GCLP),设计了一个三阶段计划,以监测该过程中每个步骤的 PBMC 完整性。第一阶段评估了基地附属实验室中新鲜 PBMC 的初始活力、总体产量和处理时间的完整性(第 1 阶段);对于第二阶段,存储库确定了从基地附属实验室收到的冷冻保存 PBMC 的解冻后活力和细胞回收率(第 2 阶段);第三阶段评估了每个存储库的长期标本储存情况(第 3 阶段)。总体而言,CHAVI PBMC QA 监督计划的结果突出了这些阶段中的每一个阶段对于从外周血采集到长期存储库保存保存标本完整性的最终目标的相对重要性。