PMID: 8601703Mar 28, 1996Paper

The cell-mediated cytotoxic response to influenza vaccination using an assay for granzyme B activity

Journal of Immunological Methods
J E McElhaneyR C Bleackley

Abstract

The measurement of cytotoxic T lymphocyte (CTL) activity through 51Cr assays is a very labour intensive method for studying cytotoxicity in human CTL due to the necessary preparation of autologous targets for the assay. An assay for granzyme B, one of a family of serine proteinases implicated in the 'lethal hit' that leads to target cell lysis, is an alternative simple measure of CTL activation. We measured granzyme B activity using its both preferred and unique substrate tert-butyloxycarbonyl-Ala-Ala-Asp-thiobenzyl ester (BAADT) in peripheral blood mononuclear cells (PBMC) obtained from influenza vaccinated subjects, and stimulated with live virus. We found that granzyme B activity increases in parallel and correlates with cytolytic activity as measured by 51Cr release assays in these virus-stimulated PBMC cultures. The assay was then used to measure the cell-mediated cytotoxic response to influenza vaccination in ten healthy elderly subjects. Peak granzyme B activity (day 6) was measured in lysates of PBMC stimulated with influenza virus, obtained from study participants before and after vaccination. We found a significant increase in granzyme B activity from pre-vaccination levels to 4 weeks post vaccination (pre=2.77 U/mg p...Continue Reading

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Citations

Aug 21, 2004·Hybridoma and Hybridomics·Jianhui RongKevin P Kane
Jul 8, 2011·Omics : a Journal of Integrative Biology·Gregory A PolandRobert M Jacobson
Sep 4, 2004·Journal of the American Veterinary Medical Association·Matthew R Sandbulte, James A Roth
Sep 24, 2013·Journal of Immunological Methods·Hannah M SalkGregory A Poland
Nov 9, 2010·Ageing Research Reviews·Janet E McElhaney
May 29, 2002·Cell Death and Differentiation·Y ZhangI N Mbawuike
Nov 5, 1998·Clinical and Diagnostic Laboratory Immunology·J E McElhaneyP Drinka

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