Foundation for National Institutes of Health, Bethesda, MD, USA.
Duke Human Vaccine Institute, Duke University, Durham, NC, USA; Duke University Medical Center, Durham, NC, USA.
J Immunol Methods. 2014 Jul;409:107-16. doi: 10.1016/j.jim.2014.04.005. Epub 2014 Apr 29.
A large repository of cryopreserved peripheral blood mononuclear cells (PBMCs) samples was created to provide laboratories testing the specimens from human immunodeficiency virus-1 (HIV-1) vaccine clinical trials the material for assay development, optimization, and validation. One hundred thirty-one PBMC samples were collected using leukapheresis procedure between 2007 and 2013 by the Comprehensive T cell Vaccine Immune Monitoring Consortium core repository. The donors included 83 human immunodeficiency virus-1 (HIV-1) seronegative and 32 HIV-1 seropositive subjects. The samples were extensively characterized for the ability of T cell subsets to respond to recall viral antigens including cytomegalovirus, Epstein-Barr virus, influenza virus, and HIV-1 using Interferon-gamma (IFN-γ) enzyme linked immunospot (ELISpot) and IFN-γ/interleukin 2 (IL-2) intracellular cytokine staining (ICS) assays. A subset of samples was evaluated over time to determine the integrity of the cryopreserved samples in relation to recovery, viability, and functionality. The principal results of our study demonstrate that viable and functional cells were consistently recovered from the cryopreserved samples. Therefore, we determined that this repository of large size cryopreserved cellular samples constitutes a unique resource for laboratories that are involved in optimization and validation of assays to evaluate T, B, and NK cellular functions in the context of clinical trials.
建立了一个大型冷冻外周血单核细胞(PBMC)样本库,为检测人类免疫缺陷病毒-1(HIV-1)疫苗临床试验标本的实验室提供了材料,用于检测开发、优化和验证。2007 年至 2013 年期间,综合 T 细胞疫苗免疫监测联盟核心库通过白细胞分离术收集了 131 份 PBMC 样本。供体包括 83 名 HIV-1 血清阴性和 32 名 HIV-1 血清阳性的受试者。使用干扰素-γ(IFN-γ)酶联免疫斑点(ELISpot)和 IFN-γ/白细胞介素 2(IL-2)细胞内细胞因子染色(ICS)检测,对这些样本进行了广泛的特征分析,以评估 T 细胞亚群对病毒抗原(包括巨细胞病毒、EB 病毒、流感病毒和 HIV-1)的反应能力。一部分样本随着时间的推移进行了评估,以确定冷冻保存样本在恢复、活力和功能方面的完整性。我们的研究的主要结果表明,从冷冻保存的样本中可以持续回收有活力和功能的细胞。因此,我们确定这个大规模冷冻细胞样本库是一个独特的资源,为参与优化和验证评估 T、B 和 NK 细胞功能的实验室提供了资源,这些实验室涉及临床试验。
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