Caspase-dependent inhibition of mousepox replication by gzmB.
BACKGROUND
Ectromelia virus is a natural mouse pathogen, causing mousepox. The cytotoxic T (Tc) cell granule serine-protease, granzyme B, is important for its control, but the underlying mechanism is unknown. Using ex vivo virus immune Tc cells, we have previously shown that granzyme B is able to activate several independent pro-apoptotic pathways, including those mediated by Bid/Bak/Bax and caspases-3/-7, in target cells pulsed with Tc cell determinants.


RESULTS
Here we analysed the physiological relevance of those pro-apoptotic pathways in ectromelia infection, by incubating ectromelia-immune ex vivo Tc cells from granzyme A deficient (GzmB(+) Tc cells) or granzyme A and granzyme B deficient (GzmAxB(-/-) Tc cell) mice with ectromelia-infected target cells. We found that gzmB-induced apoptosis was totally blocked in ectromelia infected or peptide pulsed cells lacking caspases-3/-7. However ectromelia inhibited only partially apoptosis in cells deficient for Bid/Bak/Bax and not at all when both pathways were operative suggesting that the virus is able to interfere with apoptosis induced by gzmB in case not all pathways are activated. Importantly, inhibition of viral replication in vitro, as seen with wild type cells, was not affected by the lack of Bid/Bak/Bax but was significantly reduced in caspase-3/-7-deficient cells. Both caspase dependent processes were strictly dependent on gzmB, since Tc cells, lacking both gzms, neither induced apoptosis nor reduced viral titers.


CONCLUSIONS
Out findings present the first evidence on the biological importance of the independent gzmB-inducible pro-apoptotic pathways in a physiological relevant virus infection model.
Pardo
Julián
J
Departamento Bioquímica y Biología Molecular y Celular, Facultad de Ciencias, Universidad de Zaragoza, Zaragoza, Spain. pardojim@unizar.es
Gálvez
Eva María
EM
Koskinen
Aulikki
A
Simon
Markus M
MM
Lobigs
Mario
M
Regner
Matthias
M
Müllbacher
Arno
A
D000818:Animals; D017209:Apoptosis; D037282:Calreticulin / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D020169:Caspases / Q000378:metabolism*; D004481:Ectromelia virus / Q000235:genetics*; D004482:Ectromelia, Infectious / Q000821:virology*; D005347:Fibroblasts / Q000378:metabolism*; D053804:Granzymes / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D013602:T-Lymphocytes, Cytotoxic / Q000378:metabolism* / Q000821:virology*; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D037282:Calreticulin; D051028:bcl-2-Associated X Protein; D053804:Granzymes; D053148:Caspase 3; D053179:Caspase 7; D020169:Caspases