Nat Med. 2003 Feb;9(2):231-5. Epub 2003 Jan 21.

Measuring the frequency of mouse and human cytotoxic T cells by the Lysispot assay: independent regulation of cytokine secretion and short-term killing.

Snyder JE, Bowers WJ, Livingstone AM, Lee FE, Federoff HJ, Mosmann TR.

Source

David H. Smith Center for Vaccine Biology and Immunology, University of Rochester Medical Center, New York, USA.

Abstract

Antigen-specific T cells demonstrate several potent effector functions during immune responses. Direct killing of infected cells is crucial for clearing viruses and other intracellular pathogens, but it has been difficult to measure the frequency of cytolytic cells. We have now developed a single-cell assay to measure the number of cytotoxic cells in a population, using a herpes simplex virus amplicon vector to express Escherichia coli beta-galactosidase in mouse or human target cells, and an Elispot to detect release of beta-galactosidase from killed target cells. This antigen-specific, perforin-dependent Lysispot assay has been combined with a cytokine Elispot in a two-color assay to confirm that cytotoxicity and interferon-gamma secretion are regulated independently. The simultaneous enumeration of cytokine-secreting and cytotoxic cells should be invaluable for ex vivo analysis of immune responses during infection and autoimmunity.

PMID:: 12539041; [PubMed - indexed for MEDLINE]

Publication Types, MeSH Terms, Substances, Grant Support

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Research Support, U.S. Gov't, P.H.S.

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Grant Support

R01 AI48414/AI/NIAID NIH HHS/United States

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