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Establishment and Maintenance of a PBMC Repository for Functional Cellular Studies in Support of Clinical Vaccine Trials

Abstract

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.

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References
1.
Reimann K, Chernoff M, Wilkening C, Nickerson C, Landay A . Preservation of lymphocyte immunophenotype and proliferative responses in cryopreserved peripheral blood mononuclear cells from human immunodeficiency virus type 1-infected donors: implications for multicenter clinical trials. The ACTG Immunology.... Clin Diagn Lab Immunol. 2000; 7(3):352-9. PMC: 95878. DOI: 10.1128/CDLI.7.3.352-359.2000. View

2.
Hansen S, Sacha J, Hughes C, Ford J, Burwitz B, Scholz I . Cytomegalovirus vectors violate CD8+ T cell epitope recognition paradigms. Science. 2013; 340(6135):1237874. PMC: 3816976. DOI: 10.1126/science.1237874. View

3.
Perfetto S, Chattopadhyay P, Lamoreaux L, Nguyen R, Ambrozak D, Koup R . Amine reactive dyes: an effective tool to discriminate live and dead cells in polychromatic flow cytometry. J Immunol Methods. 2006; 313(1-2):199-208. DOI: 10.1016/j.jim.2006.04.007. View

4.
Locci M, Havenar-Daughton C, Landais E, Wu J, Kroenke M, Arlehamn C . Human circulating PD-1+CXCR3-CXCR5+ memory Tfh cells are highly functional and correlate with broadly neutralizing HIV antibody responses. Immunity. 2013; 39(4):758-69. PMC: 3996844. DOI: 10.1016/j.immuni.2013.08.031. View

5.
Gill D, Huang Y, Levine G, Sambor A, Carter D, Sato A . Equivalence of ELISpot assays demonstrated between major HIV network laboratories. PLoS One. 2010; 5(12):e14330. PMC: 3001861. DOI: 10.1371/journal.pone.0014330. View