4.3 Article

OMIP-056: Evaluation of Human Conventional T Cells, Donor-Unrestricted T Cells, and NK Cells Including Memory Phenotype by Intracellular Cytokine Staining

Journal

CYTOMETRY PART A
Volume 95A, Issue 7, Pages 722-725

Publisher

WILEY
DOI: 10.1002/cyto.a.23753

Keywords

cytometry; human PBMC; T cells; MAIT cells; gamma delta T cells; NK cells; memory; intracellular cytokines

Funding

  1. Bill and Melinda Gates Foundation [OPP1066048, OPP1088952, OPP1099507]
  2. National Institute of Allergy and Infectious Diseases [P30 AI027757, UM1 AI068618]
  3. University of Washington/Fred Hutch Center for AIDS Research [P30 AI027757]
  4. Bill and Melinda Gates Foundation [OPP1088952, OPP1099507, OPP1066048] Funding Source: Bill and Melinda Gates Foundation

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A 26-color staining panel was developed to profile human antigen-specific T cells in an intracellular cytokine staining (ICS) assay using peptide pools to various antigens of interest. In addition to multiple functional markers, the panel includes differentiation/activation markers and markers to assess gamma delta, mucosal-associated invariant T, and NK T cells as well as conventional NK cells. Panel optimization was performed using previously cryopreserved PBMC from healthy adults, and then, expression of key functional markers in the panel was cross-validated against a validated ICS assay used in the HIV Vaccine Trials Network (HVTN). The panel is currently being used to evaluate the responses to tuberculosis and malaria vaccine candidates in volunteers from different geographic areas. (C) 2019 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

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