J. Exp. Med., Volume 186, Number 10, November 17, 1997 1781-1786

BRIEF DEFINITIVE REPORT:
In Vitro- and Ex Vivo-derived Cytolytic Leukocytes from Granzyme A × B Double Knockout Mice Are Defective in Granule-mediated Apoptosis but not Lysis of Target Cells

By Markus M. Simon,^* Michael Hausmann,^* Thao Tran,^* Klaus Ebnet,^* Jürg Tschopp, Ron ThaHla,^§ and Arno Müllbacher^§

From the ^* Max-Planck-Institut für Immunbiology, Stübeweg 51, D-79108 Freiburg, Germany;  Institute of Biochemistry, University of Lausanne, CH-1066 Epalinges, Switzerland; and ^§ Division of Immunology and Cell Biology, John Curtin School of Medical Research, Australian National University, Canberra City, A.C.T. 2601, Australia

Granzyme (gzm) A and gzmB have been implicated in Fas-independent nucleolytic and cytolytic processes exerted by cytotoxic T (Tc) cells, but the underlying mechanism(s) remains unclear. In this study, we compare the potential of Tc and natural killer (NK) cells of mice deficient in both gzmA and B (gzmA×B^/) with those from single knockout mice deficient in gzmA (^/), gzmB (^/), or perforin (^/) to induce nuclear damage and lysis in target cells. With the exception of perforin^/, all in vitro- and ex vivo-derived Tc and NK cell populations from the mutant strains induced ⁵¹Cr-release in target cells at levels and with kinetics similar to those of normal mice. This contrasts with their capacity to induce apoptotic nuclear damage in target cells. In gzmA×B^/ mice, Tc/NK-mediated target cell DNA fragmentation was not observed, even after extended incubation periods (10 h), but was normal in gzmA-deficient and only impaired in gzmB-deficient mice in short-term (2-4 h), but not long-term (4-10 h), nucleolytic assays. This suggests that gzmA and B are critical for Tc/NK granule- mediated nucleolysis, with gzmB being the main contributor, while target cell lysis is due solely to perforin and independent of both proteases.

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