Visualization and quantification of T cell-mediated cytotoxicity using cell-permeable fluorogenic caspase substrates

Luzheng Liu, Ann Chahroudi, Guido Silvestri, Mary E. Wernett, William J. Kaiser, Jeffrey T. Safrit, Akira Komoriya, John D. Altman, Beverly Z. Packard, Mark B. Feinberg

Research output: Contribution to journalArticlepeer-review

125 Scopus citations

Abstract

We have developed a non-radioactive flow-cytometry assay to monitor and quantify the target-cell killing activities mediated by cytotoxic T lymphocytes (CTLs). This flow-cytometry CTL (FCC) assay is predicated on measurement of CTL-induced caspase activation in target cells through detection of the specific cleavage of fluorogenic caspase substrates. Here we show that this assay reliably detects antigen-specific CTL killing of target cells, and demonstrate that it provides a more sensitive, more informative and safer alternative to the standard 51Cr-release assay most often used to quantify CTL responses. The FCC assay can be used to study CTL-mediated killing of primary host target cells of different cell lineages, and enables the study of antigen-specific cellular immune responses in real time at the single-cell level. As such, the FCC assay can provide a valuable tool for studies of infectious disease pathogenesis and development of new vaccines and immunotherapies.

Original languageEnglish (US)
Pages (from-to)185-189
Number of pages5
JournalNature Medicine
Volume8
Issue number2
DOIs
StatePublished - Feb 2002

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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