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

Nature Medicine
Luzheng LiuMark B Feinberg

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.

References

Oct 4, 1996·Science·J D AltmanM M Davis
Oct 15, 1996·Proceedings of the National Academy of Sciences of the United States of America·B Z PackardL Brand
May 2, 1998·Science·H L Ploegh
Aug 6, 1998·Journal of Immunological Methods·M M NociariC Russo
Dec 22, 1998·The Journal of Experimental Medicine·A J ZajacR Ahmed
Jun 3, 2000·Annual Review of Immunology·P C Doherty, J P Christensen
Jul 6, 2000·The Journal of Experimental Medicine·V AppayS L Rowland-Jones
Feb 28, 2001·Journal of Immunological Methods·M E SheehyD F Nixon

❮ Previous
Next ❯

Citations

Sep 24, 2004·International Journal of Cancer. Journal International Du Cancer·Yolanda D MahnkePedro Romero
Jul 20, 2002·Current Infectious Disease Reports·M. Patricia D'SouzaMargaret I. Johnston
Dec 13, 2002·Vaccine·Pedro RomeroJean-Charles Cerottini
Sep 6, 2003·Methods : a Companion to Methods in Enzymology·A ChahroudiM B Feinberg
Jan 7, 2003·Nature Medicine·Keith R JeromeMartine Aubert
Oct 18, 2005·Nature Medicine·Marie-Christine MeunierClaude Perreault
Aug 25, 2004·Proceedings of the National Academy of Sciences of the United States of America·Silvija I StapransMark B Feinberg
Jun 9, 2004·The Journal of Experimental Medicine·Ramon ArensMarinus H J van Oers
Oct 27, 2004·Journal of Translational Medicine·Nigel J WaterhouseChris Jp Clarke
Sep 26, 2013·Journal of Immunological Methods·Pauline GourdainSylvie Le Gall
Nov 1, 2002·Proceedings of the National Academy of Sciences of the United States of America·Reiko OnukiKazunari Taira
May 5, 2011·Journal of Biomedicine & Biotechnology·Shigeo KoidoHisao Tajiri
Aug 2, 2014·Proceedings of the National Academy of Sciences of the United States of America·David GateTerrence Town
Oct 29, 2003·Journal of Immunological Methods·Michael R BettsRichard A Koup
Mar 17, 2010·Clinical Immunology : the Official Journal of the Clinical Immunology Society·Shigeo KoidoHisao Tajiri
Jul 1, 2009·Immunology and Allergy Clinics of North America·Priska LochmatterWerner J Pichler
Dec 8, 2015·Journal of Neuro-oncology·Jonathan B LamanoOrin Bloch
Jan 24, 2007·Journal of Hepatology·Barbara Rehermann, Nikolai V Naoumov
Jul 17, 2009·Journal of Cellular and Molecular Medicine·Maurizio ProvenzanoGiulio C Spagnoli
Jul 19, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Carel A van BaalenGuus F Rimmelzwaan
May 9, 2008·Clinical and Experimental Immunology·P T KingS R Holdsworth
Feb 23, 2011·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·G PorebskiW J Pichler
Jul 8, 2011·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Justin PollaraGuido Ferrari
Apr 9, 2010·Hepatology Research : the Official Journal of the Japan Society of Hepatology·Jing-Yue YangWen-Chao Liu
Sep 30, 2004·International Immunopharmacology·P K Raveendran NairCheppail Ramachandran
Jul 15, 2004·Journal of Immunological Methods·Andrea WerenAndreas Ambach
Oct 6, 2005·Journal of Immunological Methods·Bettina LanghansUlrich Spengler

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.