ELISPOT assay to measure antigen-specific murine CD8(+) T cell responses


Carvalho, LH; Hafalla, JC; Zavala, F; (2001) ELISPOT assay to measure antigen-specific murine CD8(+) T cell responses. Journal of immunological methods, 252 (1-2). pp. 207-18. ISSN 0022-1759 DOI: https://doi.org/10.1016/S0022-1759(01)00331-3

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Abstract

The enzyme-linked immunospot technique (ELISPOT) relies on the visualization of cytokine secretion by individual T cells following in vitro stimulation with antigen. This assay has been developed and standardized for the quantitative detection of antigen-specific CD8(+) T cells in mice subjected to different immunization protocols [J. Immunol. Methods 181 (1995) 45]. We have identified important variables that affect the efficacy of the ELISPOT assay and in this protocol we describe this methodology in detail. As a model, we used the production of interferon-gamma by CD8(+) T cells from peripheral blood, spleen and liver of mice immunized with malaria sporozoites expressing the H-2K(d)-restricted SYVPSAEQI. This protocol has also been used successfully to detect Th1 and Th2 epitope specific CD4(+) T cells.

Item Type: Article
Keywords: Animals, Antigen-Presenting Cells, Antigens, Protozoan, CD8-Positive T-Lymphocytes, Epitopes, T-Lymphocyte, Immunoenzyme Techniques, Mice, Mice, Inbred BALB C, Plasmodium yoelii, Animals, Antigen-Presenting Cells, immunology, Antigens, Protozoan, immunology, CD8-Positive T-Lymphocytes, immunology, Epitopes, T-Lymphocyte, immunology, Immunoenzyme Techniques, methods, Mice, Mice, Inbred BALB C, Plasmodium yoelii, immunology
Faculty and Department: Faculty of Infectious and Tropical Diseases > Dept of Immunology and Infection
PubMed ID: 11334981
Web of Science ID: 168460300022
URI: http://researchonline.lshtm.ac.uk/id/eprint/9818

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