Evaluating the cytotoxicity of innate immune effector cells using the GrB ELISPOT assay

Journal of Translational Medicine
Kimberly Shafer-WeaverAnatoli Malyguine

Abstract

BACKGROUND: This study assessed the Granzyme B (GrB) ELISPOT as a viable alternative to the 51Cr-release assay for measuring cytotoxic activity of innate immune effector cells. We strategically selected the GrB ELISPOT assay because GrB is a hallmark effector molecule of cell-mediated destruction of target cells. METHODS: We optimized the GrB ELISPOT assay using the human-derived TALL-104 cytotoxic cell line as effectors against K562 target cells. Titration studies were performed to assess whether the ELISPOT assay could accurately enumerate the number of GrB-secreting effector cells. TALL-104 were treated with various secretion inhibitors and utilized in the GrB ELISPOT to determine if GrB measured in the ELISPOT was due to degranulation of effector cells. Additionally, CD107a expression on effector cells after effector-target interaction was utilized to further confirm the mechanism of GrB release by TALL-104 and lymphokine-activated killer (LAK) cells. Direct comparisons between the GrB ELISPOT, the IFN-gamma ELISPOT and the standard 51Cr-release assays were made using human LAK cells. RESULTS: Titration studies demonstrated a strong correlation between the number of TALL-104 and LAK effector cells and the number of GrB spot...Continue Reading

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Dec 31, 2003·Journal of Translational Medicine·Kimberly Shafer-WeaverAnatoli Malyguine

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Citations

Jul 8, 2005·Journal of Immunotherapy·Mark W BurkettAnatoli Malyguine
May 16, 2006·Journal of Immunotherapy·Kimberly Shafer-WeaverAnatoli Malyguine
Oct 27, 2004·Journal of Translational Medicine·Nigel J WaterhouseChris Jp Clarke
Nov 30, 2010·Journal of Immunological Methods·Theodore S JohnsonJack J Bleesing
Jun 14, 2005·Current Opinion in Immunology·Maria A SuniHolden T Maecker
Jan 1, 2012·Vaccines·Aleksandra BartnikShi-Yu Yang
Jul 11, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Kathleen M CapaccioneAndrei Molotkov
Apr 27, 2021·Contact Dermatitis·Paulina KowalczykMarian Szczepanik

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