Exp Mol Med.
2006 Feb;38(1):11-17.
Activation of the intrinsic mitochondrial apoptotic pathway in swine influenza virus-mediated cell death
- Affiliations
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- 1College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju 361-711, Korea.
- 2The Laboratory of Cell Growth and Function Regulation, Division of Bioscience and Technology, College of Life and Environmental Sciences, Korea University, Seoul 136-701, Korea. hg-kim@korea.ac.kr
- 3College of Veterinary Medicine, Chungnam National University, Yusung, Daejeon 305-764, Korea.
- 4Department of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA.
- 5Department of Animal Science, University of Minnesota, St. Paul, MN, USA.
Abstract
- The mitochondrial pathway of swine influenza virus (SIV)-induced apoptosis was investigated using porcine kidney (PK-15) cells, swine testicle (ST) cells, and HeLa cervical carcinoma cells which are known not to support viral replication. As judged by cell morphology, annexin V staining, and DNA fragmentation, PK-15 and ST cells infected with three different subtypes of SIV (H1N1, H3N2, and H1N2) were obviously killed by apoptosis, not necrosis. SIV infection in PK-15 and HeLa cells was shown to decrease the cellular levels of Bcl-2 protein compared to that of mock-infected control cells at 24 h post-infection, whereas expression levels of Bax protein increased in the PK-15 cells, but did not increase in HeLa cells by SIV infection. Cytochrome c upregulation was also observed in cytosolic fractions of the PK-15 and HeLa cells infected with SIV. Apoptosome (a multi-protein complex consisting of cytochrome c, Apaf-1, caspase-9, and ATP) formation was confirmed by immunoprecipitation using cytochrome c antibody. Furthermore, SIV infection increased the cellular levels of TAJ, an activator of the JNK-stressing pathway, and the c-Jun protein in the PK-15 and HeLa cells. Taken together, these results suggest that the mitochondrial pathway should be implicated in the apoptosis of PK-15 cells induced by SIV infection.
Keyword
MeSH Terms
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Animals
Annexin A5/metabolism
*Apoptosis
Blotting, Western
Cell Fractionation
Cell Line
Comparative Study
Cytochrome c Group/metabolism
Cytosol/chemistry
DNA Fragmentation
Enzyme Activation
Gene Expression Regulation, Viral
Hela Cells
Humans
Influenza A virus/*physiology
Kinetics
Mitochondria/metabolism/*physiology
Precipitin Tests
Proto-Oncogene Proteins c-bcl-2/genetics/metabolism
Research Support, Non-U.S. Gov't
Swine
bcl-2-Associated X Protein/genetics/metabolism
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