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Updated: Jul 11, 2026

A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl
Published on: November 28, 2014
Granzyme B production distinguishes recently activated CD8(+) memory cells from resting memory cells
Tobias M Nowacki1, Stefanie Kuerten, Wenji Zhang
1Department of Pathology, Case Western Reserve University, Cleveland, OH 44106, USA.
Abstract:
For immune diagnostic purposes it would be critical to be able to distinguish between ongoing immune processes, such as active infections, and long-term immune memory, for example imprinted by infections that have been cleared a long time ago or by vaccinations. We tested the hypothesis that the secretion of granzyme B, as detected in ex vivo ELISPOT assays, permits this distinction. We studied EBV-, flu- and CMV-specific CD8(+) cells in healthy individuals, Vaccinia virus-reactive CD8(+) cells in the course of vaccination, and HIV-specific CD8(+) cells in HIV-infected individuals. Antigen-specific ex vivo GzB production was detected only transiently after Vaccinia immunization, and in HIV-infected individuals. Our data suggest that ex vivo ELISPOT measurements of granzyme B permit the identification of actively ongoing CD8(+) cell responses-a notion that is pertinent to the immune diagnostic of infections, transplantation, allergies, autoimmune diseases, tumors and vaccine development.
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