Usage of standardized antigen-presenting cells improves ELISpot performance for complex protein antigens

Navarrete, Marcelo A.; Bertinetti-Lapatki, Cristina; Michelfelder, Ines; Veelken, Hendrik

Abstract

The enzyme-linked immunospot (ELISpot) assay is a widely used method for immune monitoring in cancer immunotherapy trials. In the ELISpot assay, peripheral blood mononuclear cells (PBMC) are stimulated with specific antigens, and cytokines of interest produced by individual cells are detected. In the standard procedure, T cells rely for antigen presentation on other cells like the monocyte/macrophage population present among the PBMC. Whereas oligopeptides can be added directly to the ELISpot assay without the necessity of a pre-incubation step, protein antigens must be internalized and processed by antigen-presenting cells to accomplish efficient presentation via HLA class I or II. We have studied the impact of sources for different antigen-presenting cell (i.e. PBMC-resident monocytes and monocyte-derived dendritic cells maturated with Poly I:C and PGE-2 based cocktails) on ELISpot assay performance and defined an optimized dendritic cell-based ELISpot protocol. This protocol is suitable for monitoring immune responses directed to protein antigens with higher sensitivity than the standard procedure. (C) 2013 Elsevier B.V. All rights reserved.

Más información

Título según WOS: ID WOS:000318467700015 Not found in local WOS DB
Título de la Revista: JOURNAL OF IMMUNOLOGICAL METHODS
Volumen: 391
Número: 1-2
Editorial: ELSEVIER SCIENCE BV
Fecha de publicación: 2013
Página de inicio: 146
Página final: 153
DOI:

10.1016/j.jim.2013.03.004

Notas: ISI