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J Immunol Methods. 2014 Jul;409:107-16. doi: 10.1016/j.jim.2014.04.005. Epub 2014 Apr 29.

Establishment and maintenance of a PBMC repository for functional cellular studies in support of clinical vaccine trials.

Author information

  • 1Foundation for National Institutes of Health, Bethesda, MD, USA.
  • 2Duke Human Vaccine Institute, Duke University, Durham, NC, USA; Duke University Medical Center, Durham, NC, USA.
  • 3Duke University Medical Center, Durham, NC, USA.
  • 4Duke Human Vaccine Institute, Duke University, Durham, NC, USA.
  • 5Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
  • 6Viral Genetics Section, US Military HIV Research Program, Henry M Jackson Foundation for the Advancement of Military Medicine, Walter Reed Army Institute of Research, Silver Spring, MD, USA.
  • 7Vaccine Research Center, NIAID, NIH, Bethesda, MD, USA.
  • 8Division of AIDS, NIAD, NIH, Rockville, MD, USA.
  • 9Becton Dickinson, San Jose, CA, USA.
  • 10International AIDS Vaccine Initiative, New York, NY, USA.
  • 11Duke Human Vaccine Institute, Duke University, Durham, NC, USA; Duke University Medical Center, Durham, NC, USA. Electronic address: gflmp@duke.edu.

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.

Copyright © 2014 Elsevier B.V. All rights reserved.

KEYWORDS:

Cryopreservation; Peripheral blood mononuclear cells; Repository

PMID:
24787274
[PubMed - indexed for MEDLINE]
PMCID:
PMC4152919
Free PMC Article
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