傳染性胰臟壞死病毒(IPNV) 是魚類的病源體，屬於典型的雙股RNA病毒。雙股RNA病毒表現兩段式雙股的RNA基因體並且包含在一個中型、不具外套膜、20面體的外殼蛋白中。
本論文中，我們發現當IPNV感染CHSE-214 6-8小時間，會引起粒線體通透性的改變，接著引起initiator Caspase-9以及effector caspase-3的活化。而IPNV感染造成宿主細胞粒線體膜電位的消失，可以藉著處理粒線體過渡性通透孔(MPTP)抑制劑BKA(20μg/ml)來抑制膜電位的消失。接著更進一步發現抗凋亡蛋白IPNV VP5能抑制IPNV經由粒線體所造成之細胞死亡。我們也發現當IPNV感染宿主細胞之後，會導致宿主細胞產生內質網壓力，並且使GRP 78基因表現量上升。
於本論文研究結果中，IPNV能經由粒線體之凋亡途徑造成宿主細胞死亡，進而誘導caspase-9與caspase-3之活化，這是相當重要之發現，進由此發現希望能對IPNV之感染與治療有所幫助。

Infectious pancreatic necrosis virus (IPNV) is a fish-derived pathogen and is the prototype of the Birnaviridae virus family. Birnaviruses possess a bi-segmented, double-stranded RNA genome contained within a medium-sized, unenveloped, icosahedral capsid.
In this thesis, we found that IPNV infection can induce the mitochondrial permeability change between 6h and 8 h post-infection (p.i.) time, which also corresponded to activate the initiator caspase-9 and effector caspase-3 for triggering cell death. Then IPNV-induced loss of mitochondrial membrane potential (MMP) can be inhibited by treatment with mitochondrial permeability transition pore inhibitor Bongkrek acid (20ug/ml) (BKA). Furthermore, the viral anti-apoptotic member IPNV VP5 can block IPNV-induced mitochondria- mediated cell death in CHSE-214 cells. Moreover, the IPNV-infection CHSE-214 can up-regulate the ER stress marker GRP 78, at middle apoptotic stage, in CHSE-214 cells.
Taken our results suggest that IPNV induces host cell apoptosis may through mitochondria-mediated death pathway, this finding may provide an important insight into the control of IPNV infection.

官柏志. 傳染性胰臟壞死病毒感染大眼鮭魚胚胎細胞株引發轉錄因子NF-kB之活化以造成細胞凋亡。台灣大學漁業科學研究所碩士論文。台北 (2001)。

黃玲娟. 傳染性胰臟壞死病毒感染斑馬魚肝細胞株誘發活化Caspases之細胞凋亡。台灣海洋水產生物科技研究所碩士論文。基隆 (2000)。

戴君如 以傳染性胰臟壞死病毒之VP3反意核醣核酸表現抑制本病毒所誘發之細胞凋亡。台灣大學漁業科學研究所碩士論文。台北(2000)。

周宜黎. 傳染性胰臟壞死病毒凋亡基因vp3所誘發細胞凋亡分子機制
之研究。成功大學生物科技研究所碩士論文。台南 (2004)。

邱謙禮. 傳染性胰臟壞死病毒外殼蛋白VP3造成斑馬魚肝臟細胞株凋亡分子機制之研究。成功大學生物科技研究所碩士論文。台南(2006)

Anne R., Richard C. M. and Philip E. B. Viruses and apoptosis. Annu. Rev. Microbiol. 53: 577-628. (1999).

Braakman, I., Hoover-Litty, H., Wagner, K. R. and Helenius, A. Folding of influenza hemagglutinin in the endoplasmic reticulum. J. Cell Biol. 114:401-411. (1991).

Brown D. G., Sun X. M. and Cohen G. M. Dexamethasone-induced apoptosis involves cleavage of DNA to large fragments prior internucleosomal fragmentation. J. Biol. Chem. 268:3037-3039. (1993).

Chen C. C., Wang J. K. and Lin S. B. Antisense oligonucleotide targeting protein kinase C-α , -β I, or-δ but not-η inhibit lipopolysaccharide -induced nitric oxide synthase expression in RAW 264.7 macrophages: involvement of a nuclear factor kappa B-dependent mechanism. J. Immunol. 161:6206-6214. (1998).

Chen S. N., Chi S. C., Guu J. J., Chen J. C. and Kou G. H. Pathogenicity of birnavirus isolated from loach CoA Fisheries series No. 10. Fish Disease Res. 6: 6-11. (1984).

Desagher S. and Martinou J. C. Mitochondria as the central control point of apoptosis. Trends Cell Biol. 10(9): 369-377. (2000).

Dobos P. Size and structure of the genome of infectious pancreatic necrosis virus. Nucl.Acids Res. 3: 1903-1924. (1976).

Dobos P. Virus-specific protein synthesis in cells infected by infectious pancreatic necrosis virus. J. Virol. 21:242-258. (1977).

Dobos P., Hallett R., Kells D. T., Sorensen O. and Rowe, D. Biophysical studies of infectious pancreatic necrosis virus. J. Virol. 22:150-159. (1997).

Dobos P., Hill B. J., Hallett R., Kells D. T. C., Becht H. and Teninges D. Biophysical and biochemical characterization of five animal viruses with bisegmented double-stranded RNA genomes. J. Virol. 32:593-605. (1979).

Dobos P. and Rowe D. Peptide map comparison of infectious pancreatic necrosis virususpecific poly-peptides. J. Virol. 24:805-820. (1977).

Duncan R., Nagy E., Krell P. J. and Dobos P. Synthesis of the infectious pancreatic necrosis virus polyproteinm detedtion of a segment A coin-gregion. J. Virol. 61:3655-3664. (1987).

Egger L., Madden D. T., Rhme C., Rao R. V., Bredesen D E. Endoplasmic reticulum stress-induced cell death mediated by the proteasome. Cell Death Differ. 14:1172–1180. (2007)

Eick D. and Hermeking H. Viruses as pacemakers in the evolution of defence mechanisms against cancer. Trends Genet. 12: 4-6. (1996).

Fan T. J., Han L. H., Cong R. S. and Liang J. Caspase family proteases and apoptosis. Acta. Biochim. Biophys. Sin. (Shanghai). 37:719-27. Review. (2005).

Gething M. J. and Sambrook J. Protein folding in the cell. Nature 355, 33-45.(1992).

Green D. R. and Reed J. C. Mitochondria and apoptosis. Science 281: 1309-1312. (1998).

Guido, K., Bruno, D. and Michele, R. R. The mitochondrial death/life regulator in apoptosis and necrosis. Annu. Rev. Physiol. 60:619-42. (1998).

Harrington E. A., Bennett M. R., Fandini A. and Evan G. I. Modulation of myc–induced apoptosis by specific cytokines. EMBO J. 13: 3286- 3295. (1994).

Hedrick R. P., Fryer J. L., Chen S. N. and Kou G. H. Characteristics of four birnavirus isolated from fish in Taiwan. Fish Pathol. 18: 91-97. (1983).

Hill B. J. and Way K. Serological classification of fish and shellfish birnaviruses. In abstract of Fish Interactional Conference of the European Association of Fish pathologists. pp. 10. (1983).

Hong J. R., Lin T. L., Hsu Y.L. and Wu J. L. Induction of apoptosis and secondary necrosis by infectious pancreatic necrosis virus in fish embryonic cells. J. Fish. Diseases. 21:463-467. (1998a).

Hong J. R., Lin T. L., Hsu Y.L. and Wu J. L. Apoptosis precedes necrosis of fish cell line with infectious pancreatic necrosis virus infection. Virology. 250: 76-84. (1998b).

Hong J. R., Hsu Y. L. and Wu J. L. Infectious pancreatic necrosis virus induces apoptosis due to down-regulation of survival factor Mcl-1 protein expression in a fish cell line. Virus. Research. 63:75-83. (1999a).

Hong J. R. and Wu J. L. Cloning and functional analysis of IPNV VP5: a novel Bcl-2 family with regulation of viral expression and anti-cell death activity. Virology. (submitted) . (2001a).

Hong J. R. and Wu J. L. Induction of apoptosis death in cell via Bad gene expression by infectious pancreatic necrosis virus infection. Cell Death Differ. 9:113-124. (2002).

Hong J. R. and Wu J. L. Molecular regulation of cellular apoptosis by fish infectious pancreatic necrosis virus infection. Virology.2:151-160. (2002).

Hong J. R. and Wu J. L. VP5, a novel anti-apoptosis gene of the Bcl-2 family, regulates Mcl-1 and viral protein expression. Virology 295(2): 217-29. (2002).

Hong J. R., Huang L.J., and Wu J. L. Aquatic birnavirus induces apoptosis through activated caspase-8 and -3 in a zebrafish cell line. J. Fish Dis. 28(3):133-40. (2005).

Hsu Y. L., Chen B. S. and Wu J. L. Comparison of RNAs and polypeptides of infectious pancreatic necrosis virus from eel and rainbow trout. J. Gen. Virol. 70: 2233-2239. (1989).

Hu Y., Benedict M. A., Ding L. and Nunez G. Role of cytochrome c and dATP / ATP hydrolysis in Apaf-1-mediated caspase-9 activation and apoptosis. EMBO Journal.18:3586-3595. (1999).

Neet K. and Hunter T. Vertebrate non-receptor protein-tyrosine kinase families. Genes Cells. 1:147-169. Review. (1996).

Kerr J. F. R. Neglected pooprtunities in apoptosis research. Trend. Cell. Biol. 5:55-57. (1995).

Kerr J. F. R., Wyllie A. H., and Currie A. R. Apoptosis : a basic biological phenomenon with wide ranging implications in tissue kinetics. Br. J. cancer. 26:1790-1794. (1972).

Kroemer G. The proto-oncogene Bcl-2 and its role in regulation apoptosis. Nature Med. 3: 614-620. (1997).

Li P. and Ahmad M. Cytochrume c and Datp-dependent formation of Apaf-1/caspase-9 complex initiates an apoptic protease cascade. Cell. 91:479-89. (1997).

Liu M., Vakharia V. N. Nonstructural protein of infectious bursal disease virus inhibits apoptosis at the early stage of virus infection. J Virol. 80:3369-77. (2006).

Liu Y., Pu Y. and Zhang X. Role of the mitochondrial signaling pathway in murine coronavirus-induced oligodendrocyte apoptosis. J Virol. 80:395-403. (2006).

Lo C. F., Hong Y. W., Huang S. Y. and Wang C. H. The characteristics of the virus isolated from the gill of clam, Meretrix lusoria. Fish Pathol. 23: 147-154. (1988).

Martins L. M. and Earnshaw W. C. Apoptosis: alive and kicking in 1997. Trends in Cell Biology. 7:111-114. (1997).

MacDonald R. D. and Gower D. A. Genomic and phenotypic divergence among three serotypes of aquatic birnaviruses (infectious pancreatic necrosis virus). Virology. 114:187-195. (1981).

Manning D. S., mason C. L. and Leong J. C. Cell-free translation analysis of the processing of infectious pancreatic necrosis virus polyprotein. Virology. 179:9-15. (1990).

Morgan M. M., Maereadie I. G., Hazley V. R., Hudson P. J. and Azad A. A. Sequence of the small double-stranded RNA genome segment of infections bursal disease virus and its deduced 90-Kda product. Virology. 163:240-242. (1997).

Kumar S. and Harvey N. L. Role of multiple cellular protease in the execution of programmed cell death. FEBS Lett. 375: 169-173. (1995).

Oldstone M. B. How viruses escape from cytotoxic T lymphocytes : molecular parameters and players. Virology 234: 179-185. (1997).

Santi N., Sandtro A., Sindre H., Song H., Hong J. R., Thu B., Wu J. L., Vakharia V. N., Evensen O. Infectious pancreatic necrosis virus induces apoptosis in vitro and in vivo independent of VP5 expression. Virology. 342:13-25. (2005).

Santi N., Sandtro A., Sindre H., Song H., Hong J. R., Thu B, Wu J. L., Vakharia V. N., Evensen O. Infectious pancreatic necrosis virus VP5 is dispensable for virulence and persistence. J Virol. 79: 9206-9216. (2005).

Saleh A., Srinivasuls S. N., Acharya S., Fishel R. and Alnemri E. S. Cytochrome d and dATP/ mediated oligomerization of Apaf-1 is a prerequisite for procaspase-9 activation. J. Biol. Chem. 274:17941-17945. (1999).

Samali A., Gorman A. M., and Cotter G. Apoptosis -- the story so far….. Experientia. 52(10-11): 933-941. (1996).

Schlissel M. S., and Baltimore D. Activation of immunoglobulin kappa gene rearrangement correlates with induction of germline kappa gene transcription. Cell. 58:1001-1007. (1989).

Sen R., and Baltimore D. Multiple nuclear factors ineract with the immunoglobulin enhancer sequences. Cell. 46:705-716. (1986).

Shimizu S., Konishi A., Kodama T. and Tsujimoto Y. BH4 domain of antiapoptotic Bcl-2 family members closes voltage-dependent anion channel and inhibits apoptotic mitochondrial changes and cell death. Proc. Natl. Acad. Sci. USA. 97: 3100-3105. (2000).

Shimizu S., Narita M. and Tsujimoto Y. Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature. 399: 483-487. (1999).

Snieszk S. F., Wolf K., Camper J. E. and Pettijohn L. L. Infections nature of pancreatic necrosis. Trans. Am. Fish Soc. 88:289-293. (1959).

Takashi M. Caspases involved in ER stress-mediated cell death. J. Chem. Neuroanat. 28:101-105. (2004).

Tewari M., Quan L. T., O'Rourke K., Desnoyers S., Zeng Z., Beidler D. R., Poirier G. G., Salvesen G. S., Dixit V. M. Yama/CPP32 beta, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase. Cell. 81:801-9. (1995).

Thornberry N. A. and Lazebnik Y. Caspases: enemies within. Science 281: 1312-1316. (1998).

Underwood B. O., Smale C. J., Brown F. and Hill B. Relationship of a virus from Tellina tenuis to infectious pancreatic necrosis virus. J. Gen. Virol. 36:93-109. (1977).

Urano F., Wang X., Bertolotti A., Zhang Y., Chung P., Harding H. P. and Ron D. Coupling of stress in the ER to activation of JNK protein kinases by transmembrane protein kinase IRE1. Science 287: 664-666. (2000).

Vaux D.L. and Strasser A. The molecular biology of apoptosis. Pro. Nat. Acad. Sci. USA. 933: 2239-2244. (1996).

Wolf, K., Snieskol, S. F., Dumbar, C. E. and Pyle, E. Virus nature of IPN in trout. Pro. Soc. EXP. Biol. Med. 104: 105-108. (1960).

Wolf K. and Mann J. A. Pokilother vertebrate cell line and virus: a current listing for fish. Virology 16: 168-179. (1980).

Wood E. M., Snieskol S. F. and Yasutake W. T. Infectious pancreatic necrosis Virus. A. M. A. Arch. Paath. 60:26-28. (1955).

Wu J. L., Chang C. Y. and Hsu Y. L. Characteristics of an infectious pancreatic necrosis like virus isolated from Japanese eel (Amguolla japonica). Bulletin of the Institute of Zoology. Academia Sinica. 26:210-214. (1987).

Yao K., Vakharia V. N. Induction of apoptosis in Vitro by the 17-kDa nonstructural protein of infectious bursal disease virus: possible role in viral pathogenesis. Virology. 1:50-8. (2001)