Ex Vivo Analysis of Human Antigen-Specific CD8+ T-Cell Responses: Quality Assessment of Fluorescent HLA-A2 Multimer and Interferon-[gamma] ELISPOT Assays for Patient Immune Monitoring.
Speiser, Daniel E. *; Pittet, Mikael J. *; Guillaume, Philippe [S]; Lubenow, Norbert +; Hoffman, Eric ++; Cerottini, Jean-Charles *[S]; Romero, Pedro *||
Journal of Immunotherapy.
27(4):298-308, July/August 2004.
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colon; The authors developed a standardized approach for immune monitoring of antigen-specific CD8 T cells within peripheral blood lymphocytes (PBLs) that combines direct ex vivo analysis of Melan-A/MART-1 and influenza-specific CD8 T cells with HLA-A2/peptide multimers and interferon-[gamma] ELISPOT assays. Here the authors assessed the quality of results obtained with 180 PBLs from healthy donors and melanoma patients. Reproducibility of the multimer assay was good (average of 15% variation). In the absence of in vivo antigen-specific T-cell responses, physiologic fluctuations of multimer-positive T cells was low, with variation coefficients of 20% for Melan-A and 28% for influenza-specific T cells. In contrast, patients with vaccination-induced T-cell responses had significantly increased T-cell frequencies clearly exceeding physiologic fluctuations. Comparable results were obtained with ELISPOT assays. In conclusion, this approach is well suited to assess T-cell responses as biologic endpoints in clinical vaccine studies.
(C) 2004 Lippincott Williams & Wilkins, Inc.