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

doi:10.1038/nm0202-185

PubMed · 11821904

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

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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.

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Animals Caspases Chromium Radioisotopes Cytotoxicity, Immunologic Female Flow Cytometry Mice Mice, Inbred C57BL Substrate Specificity T-Lymphocytes, Cytotoxic Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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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. 2002. Visualization and quantification of T cell-mediated cytotoxicity using cell-permeable fluorogenic caspase substrates.. https://doi.org/10.1038/nm0202-185

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