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

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

Research output: Contribution to journalArticlepeer-review

56 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)207-218
Number of pages12
JournalJournal of Immunological Methods
Issue number1-2
StatePublished - Jun 1 2001
Externally publishedYes


  • CD8 T cells
  • Interferon-γ

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology


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