A reproducible method for the enumeration of functional (cytokine producing) versus non-functional peptide-specific cytotoxic T lymphocytes in human peripheral blood

S. N. Markovic, W. K. Nevala, C. B. Uhl, E. Celis, D. J. McKean

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

One of the most difficult laboratory challenges in the field of therapeutic cancer vaccines has been the development of uncomplicated/reproducible methods for the quantification of vaccine immunization efficacy in peripheral blood of cancer patients. Existing methods are limited by lack of functional information (tetramers), difficulties with standardization/reproducibility [enzyme-linked immunosorbent spot (ELISPOT)] and reliance on endogenous (sample-specific) antigen presentation (cytokine flow cytometry). Herein we present a reproducible method utilizing an artificial antigen-presenting cell platform for flow cytometry-based quantification of the frequency and activation status of peptide-specific cytotoxic T lymphocytes. The methodology [currently presented for cytomegalovirus human leucocyte antigen (HLA)-A2 cognant peptide antigens] allows simultaneous ex vivo quantification of activated (cytokine-producing) and inactive tetramer-positive T cells following HLA class I/peptide/CD28 stimulation independent of endogenous antigen presentation. The simplicity and reliability of the assay provide for high-throughput applications and automation. The utility and application of this method are discussed.

Original languageEnglish (US)
Pages (from-to)438-447
Number of pages10
JournalClinical and Experimental Immunology
Volume145
Issue number3
DOIs
StatePublished - Sep 1 2006
Externally publishedYes

Keywords

  • Artificial antigen-presenting cells
  • Cytomegalovirus
  • Immunity
  • T cells
  • Tolerance

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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