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

Luzia H. Carvalho; Julius C.R. Hafalla; Fidel Zavala

doi:10.1016/S0022-1759(01)00331-3

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

Luzia H. Carvalho, Julius C.R. Hafalla, Fidel Zavala

Research output: Contribution to journal › Article › peer-review

56 Scopus citations

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-γ 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.

Original language	English (US)
Pages (from-to)	207-218
Number of pages	12
Journal	Journal of Immunological Methods
Volume	252
Issue number	1-2
DOIs	https://doi.org/10.1016/S0022-1759(01)00331-3
State	Published - Jun 1 2001
Externally published	Yes

Keywords

CD8 T cells
ELISPOT
Interferon-γ

ASJC Scopus subject areas

Immunology and Allergy
Immunology

Access to Document

10.1016/S0022-1759(01)00331-3

Cite this

@article{ce8454db972f4be8beaf78dd749eb66d,

title = "ELISPOT assay to measure antigen-specific murine CD8+ T cell responses",

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-γ by CD8+ T cells from peripheral blood, spleen and liver of mice immunized with malaria sporozoites expressing the H-2Kd-restricted SYVPSAEQI. This protocol has also been used successfully to detect Th1 and Th2 epitope specific CD4+ T cells.",

keywords = "CD8 T cells, ELISPOT, Interferon-γ",

author = "Carvalho, {Luzia H.} and Hafalla, {Julius C.R.} and Fidel Zavala",

note = "Funding Information: This work was supported by NIH grant AI44375. L.H.C. is a fellow of the Brazilian Research Council (CNPq). We thank Dr. Gloria Gonzalez-Aseguinolaza for critical review of the manuscript.",

year = "2001",

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doi = "10.1016/S0022-1759(01)00331-3",

language = "English (US)",

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T1 - ELISPOT assay to measure antigen-specific murine CD8+ T cell responses

AU - Carvalho, Luzia H.

AU - Hafalla, Julius C.R.

AU - Zavala, Fidel

N1 - Funding Information: This work was supported by NIH grant AI44375. L.H.C. is a fellow of the Brazilian Research Council (CNPq). We thank Dr. Gloria Gonzalez-Aseguinolaza for critical review of the manuscript.

PY - 2001/6/1

Y1 - 2001/6/1

N2 - 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-γ by CD8+ T cells from peripheral blood, spleen and liver of mice immunized with malaria sporozoites expressing the H-2Kd-restricted SYVPSAEQI. This protocol has also been used successfully to detect Th1 and Th2 epitope specific CD4+ T cells.

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KW - Interferon-γ

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