(18.206.238.77) 您好!臺灣時間:2021/05/17 17:47
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果

詳目顯示:::

我願授權國圖
: 
twitterline
研究生:鄒宗萱
研究生(外文):Tsung-Hsiuan Tsou
論文名稱:人類多瘤性病毒JCV重組殼體
論文名稱(外文):Study of JC Virus Pseudocapsid Infection to Human Cells
指導教授:許國堂張德卿
指導教授(外文):Gwo-Tarng SheuDeching Chang
學位類別:碩士
校院名稱:中山醫學院
系所名稱:毒理學研究所
學門:醫藥衛生學門
學類:其他醫藥衛生學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:62
中文關鍵詞:多瘤性病毒基因治療
外文關鍵詞:polyomavirusgene therapy
相關次數:
  • 被引用被引用:0
  • 點閱點閱:230
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
中 文 摘 要
人類多瘤性病毒JC病毒主要結構蛋白VP1可在大腸桿菌系統中表達,並能自行組裝成殼體。利用10-50%蔗糖濃度梯度離心,可純化出VP1空殼體(capsid-like particle)。本實驗室先前已經證明此VP1空殼體可包裝外生性DNA,並運送螢光基因到人類腎臟細胞(293)中表達,顯示此VP1空殼體可能可以作為人類基因治療之載體。本研究則是將純化的VP1空殼體利用滲透壓震盪(osmotic shock)的方式在in vitro環境下包裝修飾的SV40大T抗原基因反義核酸(SV40LTASMODN),其空殼體與ODN包裝比例為1:132。含ODN殼體感染猴子腎臟細胞(COS-7),除了發現在COS-7內SV40 LT蛋白質的表達有明顯的下降外,SV40LTASMODN亦會造成COS-7細胞走向細胞凋亡(apoptosis)的路徑。以上的結果顯示,VP1空殼體的確可以攜帶ODN進入細胞,並且能發揮生物功能,抑制細胞內SV40LT之mRNA的轉譯。此外,亦利用SV40LTASMODN-FITC包入VP1空殼體內,作為感染人類細胞後偵測被感染細胞的指標,結果發現人類腦細胞(98﹪)、腎臟細胞(82%)、肺細胞(74%)、骨細胞(51%)、皮膚細胞(60%)、膀胱細胞(18%)等,皆可被JC病毒所感染,但感染效率並不相同;另外食道細胞、乳腺細胞、T淋巴球、胃細胞以及直腸細胞,皆不能被JC病毒所感染。此項研究的結果,對於未來以人類多瘤性病毒JCV作為臨床基因治療載體的研究及應用提供了相當重要的訊息。

Abstract
The major capsid protein VP1 of human polyomavirus, JC virus, has been expressed in E. coli and self-assembled into a virus-like particle (VLP) structure. The VLP has been purified by 10-50% sucrose gradient centrifugation. The VLPs were further demonstrated to package and deliver exogenous DNA into human kidney cells. These findings indicate that the recombinant JCV capsid potentially could be used as a human gene transfer vector. To further explore the potential of the recombinant VLP gene therapy, antisense oligodeoxynucleotide (ODN) was packaged into the VLP by osmotic shock. The package ratio of the VLP to ODN was approximately 1 to 132. The antisense ODN against SV40 large T antigen (SV40LTASMODN-FITC) was delivered into the cytoplasm of COS-7 cells by JCV VLP. Expression of large T antigen was inhibited. In addition, apoptosis was also found after the treatment. These results indicate that the JC VLP could be developed as a gene transfer vector for gene therapy. Finally, susceptibility of various human cell lines to JCV-VLP was investigated. The results showed that human astrocytoma(98%), brain(98%), kidney(82%), lung(74%), bone(51%), bladder(18%), and skin(60%) cell can be targeted by JCV recombinant capsid. Further investigation is required to construct the recombinant VLP, and our findings will provide valuable groundbreaking data to optimize the requirements of using JC-VLP for gene delivery an alternative method of gene therapy.

目 錄
頁次
中文摘要 1
英文摘要 2
前言
人類多瘤性病毒背景介紹3
本研究相關背景介紹 6
材料與方法 13
結果 25
討論 33
參考文獻 38
圖表 47

參 考 資 料
1.Padgett B. L., G. ZuRhein D. L. Walker R. Echroade and B. Dessel. 1971. Cultivation of papova-like virus from human brain with progressive multifocal leukoencephalopathy . Lancet I :1257-1260.
2.Astrom KE, Mancall EL, Richardson EP Jr. 1958. Progressive mutifocal leukoencephalopathy. Brain. 81:93-127.
3.Gardner S. D., A. M. Field, D. V. Coleman, and B. Hulme.1971. New human papovavirus (BK) isolated from urine after renal transplantation. Lancet I :1253-1257
4.Padgett B. L. and D. L. Walker. 1973. Prevalence of antibodies in human sera against JC virus, an isolate from a case of progressive multifocal leukoencephalopathy. J. Infect. Dis. 127:467-470.
5.Brown P., T. Tsai, and D. C. Gajdusek. 1975. Seroepidemiology of human papovavirus: discovery of virgin population and some unusual patterns of antibody prevalence among remote peoples of the world. Am. J. Epidemiol. 102:331-340.
6.Rziha H. J., G. Bornkamm, and H. ZurHausen. 1978. Seroepidemiological studies and serologic response to viral infection. Med. Microbiol. Immunol. 165:73-92.
7.Chesters P. M., Heritage J., McCance D. J. 1983. Persistence of DNA sequences of BK virus and JC virus in normal human tissues and in diseased tissues. Journal of Infectious Disease. 147:676-682.
8.Dorries K. 1984. Progressive multifocal leucoencephalopathy: analysis of JC virus DNA from brain and kidney tissue. Virus Research. 1:25-38.
9.Kitamura T, Sugimoto C, Kato A, Ebihara H, Suzuki M, Taquchi F, Kauabe K, Yogo Y. 1997. Persistent JC virus (JCV) infection is demonstrated by continuous shedding of the same JCV strains. Journal of Clinical Microbiology. 35:1255-1257.
10.Arthur R. R., Shan K. V., Charache P., Saral R. 1989. BK and JC virus infections in recipients of bone marrow transplants. Journal of Infectious Disease. 158:563-569.
11.Chang D., Wang M., Ou W. C., Lee M. S., Ho H.N., Tsai R. T. 1996. Genotypes of human polyomaviruses in urine samples of pregnant individuals in Taiwan. Journal of Medical Virology. 48:95-101.
12.Chang D., Tsai R. T., Wang M., Ou W. C. 1996. Different genotypes of human polyomaviruses found in patients with autoimmune diseases in Taiwan. Journal of Medical Virology. 48:204-209.
13.Coleman D. V., Wolfendale M. R., Daniel R. A. 1980. A prospective study of human polyomavirus infection in pregnancy. Journal of Infectious Disease. 142:1-8.
14.Gardner S., Mackenzie E., Smith C., Porter A. 1984. Prospective study of the human polyomavirus BK and JC and cytomegalovirus in renal transplant recipients. Journal of Clinical Pathology. 37:578-586.
15.Myers C., Frisque R. J., Arthur R. R. 1989. Direct isolation and characterization of JC virus from urine samples of renal and bone marrow transplant patients. Journal Virology. 63:4445-4449.
16.Tsai R. T., Wang M., Ou W. C., Lee Y. L., Li S. Y., Fung C. Y., Huang Y. L., Tzeng T. Y., Chang D. 1997. Viruria incidence of JC virus is higher than that of BK virus in Taiwan. Journal of Medical Virology. 52:253-257.
17.Yogo Y., Kitamura T., Sugimoto C., et al. 1990. Isolation of a possible archetypical JC virus DNA sequence from nonimmunocompromised individuals. J Virol; 64: 3139-43.
18.Padgett B. L., Walker D. L. 1973. Prevalence of antibodies in human sera against JC virus, and isolate from a case of progressive multifocal leukoencephalopathy. Journal of Infectious Disease. 127:467-470.
19.Padgett B. L., Walker D. L., ZuRhein G. M., Hodach A. E., Chow S. M. 1976. JC papovavirus in progressive multifocal leukoencephalopathy. Journal of Infectious Disease. 133:686-690.
20.Padgett B. L., Rogers G. M., Walker D. L. 1977. JC virus, a human polyomavirus associated with progressive multifocal leukoencephalopathy: additional biological characteristics and antigenic relationships. Infectious Immunology. 15:656-662.
21.Gallia G. L., Houff S. A., Major E. O., Khalili K. 1997. Review: JC virus infection of lymphocytes-revisited. Journal of Infectious Disease. 176:1603-1609.
22.Lafon M. E., Dutronc H., Dubois V., Pellegrin I., Barbeau P., Ragnaud J. M., Pellegrin J. L., Fleury N. J. 1998. JC virus remains latent in peripheral blood B-lymphocytes but replicates actively in urine from AIDS patients. Journal of Infectious Disease. 177:1502-1505.
23.Monaco M. C., Jensen P. N., Hou J., Durham L. C., Major E. O. 1998. Detection of JC virus DNA in human tonsil tissue: evidence for site of initial viral infection. Journal of Virology. 72:918-923.
24.Tornatore C., Berger J. R., Houff S. A., Curfman B., Meyers K., Winfield D., Major E. O. 1992. Detection of JC virus DNA in peripheral lymphocytes from patients with and without progressive multifocal leukoencephalopathy. Annal of Neurology. 31:454-462.
25.Dubois V., Dutronc H., Lafon M. E., Poinsot V., Pellegrin J. A. L., Ragnand J. M., Ferrer A. M., Fleury H. J. 1997. Latency and reactivation of JC virus in peripheral blood of human immunodeficiency virus type I-infected patients. Journal of Clinical Microbiology. 35:2288-2292.
26.Major E. O., Amemiya K., Tornatore C. S., Houff S. A., Berger J. R. 1992. Pathogenesis and molecular biology of Progressive Multifocal Leukoencephalopathy, a JC Virus-induced demyelinating disease of the human brain. Clinical Microbiology Review. 5:49-73.
27.Howley P. M. 1980. Molecular biology of SV40 and the human polyomaviruses BK and JC. In: G. Klein (ed). Viral Oncology. Ravan Press. New York, 1980: 489-550.
28.Frisque R. J., Bream G. L., Canella M. T. 1984. Human polyomavirus JC virus genome. Journal of Virology 51:458-469.
29.Frisque R. J., G. Bream, and M. Cannellla. 1984. Human polyomavirus JC virus genome. J. Virol. 51:458-469.
30.Carswell S., Alwine J. C. 1986. Simian virus 40 agnoprotein facilitates perinuclear localization of VP1, the major capsid protein. Journal of Virology. 60:1055-1061.
31.Gharakhanian E., Takahashi J., Clever J., Kasamatsu H. 1988. In vitro assay for protein-protein interaction: Carboxyl-terminal 40 residues of simian virus 40 structural protein VP3 contain a determinant for interaction with VP1. Proc. Natl. Acad. Sci. USA. 85:6607-6611.
32.Anderson W. F. 1992. Human gene therapy. Science. 256: 808-813.
33.Cosset F. L., and S. J. Russell. 1996. Targeting retrovirus entry. Hum. Gene Ther. 3:946-956.
34.Forstova. J., N. Krauzewicz, V. Sandig, J. Elliott, Z. Palkova, M. Strauss, and B. E. Griffin. 1995. Polyoma virus pseudocapsids as efficient carriers of heterologous DNA into mammalian cells. Hum. Gene Ther. 6: 297-306.
35.Anderson W. F. 1998. Human gene therapy. Nature. 392:25-30.reviews.
36.Kay M. A., D. Liu, and P. M. Hoogerbrugge. 1997. Gene therapy. Proc. Natl. Acad. Sci. USA. 94:12744-12746.
37.Breakefield X.O., and N. A. DeLuca. 1991. Herpes simplex virus for gene delivery to neurons. New Biol. 3:203-218.
38.Fink D. J., and J. C. Glorioso. 1997. Engineering herpes simplex virus vectors for gene transfer to neurons. Nature Med. 3: 357-359
39.Bilba G. 1997. Adenoviral/retroviral vector chimeras: a novel strategy to achieve high-efficiency stable transduction in vivo. FASEB J. 11:624-634
40.Ledley F. D. 1995. Nonviral gene Therapy: the promise of genes as pharmaceutical products. Hum. Gene Ther. 6:1129-1144.
41.Osterman JV, Waddell A, and Aposhian HV 1970. DNA and gene therapy: uncoating of polyoma pseudovirus in mouse embryo cells.
Proc Natl Acad Sci USA. Sep;67(1):37-40.
42.Kashmiri S.V.S., and Aposhian H.V. 1974. Degradation of pseudoviral DNA after infection of mouse cells with polyoma pseudovirions. Proc. Natl. Acad. Sci. USA. 71:3834-3838.
43.Forstova J., Krauzewicz N., Elliott J., Palkova Z., Strauss M., and Griffin B.E. 1995. Polyoma virus pseudocapsids as efficient carriers of heterologous DNA into mammalian cells. Human Gene Therapy.6:297-306.
44.Barr S.M., Keck K., and Aposhian H.V. 1979. Cell-free assembly of a polyoma-like particle from empty capsids and DNA. Virology. 96:656-659.
45.Kawana K., H. Yoshikawa, Y. Taketani, K. Yoshike, and T. Kanda. 1998. In vitro construction of pseudovirions of human papillomavirus type 16: Incorporation of plasmid DNA into reassembled L1/L2 capsids. J. Virol. 72:10298-10300.
46.Goldmann C., P. Harald, F. Stephan, A. Oliver, E. Susanne, J. Klaus-dieter, K. Franz-Josef, W. Frank, T. Corinna, N.Thomas, H. Gerhard, W. Thomas and L. Wolfgang. 1999. Molecular cloning and expression of major structural protein VP1 of the human polyomavirus JC virus: formation of virus-like particles useful for immunological and therapeutic studies. J. Virol. 73:4465-4469.
47.Stephenson M. L., and Zamecnik P. C. 1978. Inhibition of Rous sarcoma viral RNA translation by a specific oligodeoxyribonucleotide. Proc Natl Acad Sci USA. 75(1):285-288.
48.Zamecnik P. C., and Stephenson M. L. 1978. Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide. Proc Natl Acad Sci USA. 75(1):280-284.
49.Willnow T. E., and Herz J. 1994. Homologous recombination for gene replacement in mouse cell lines. Methods Cell Biol. 43 Pt A:305-334.
50.Melton D. W. 1994. Gene targeting in the mouse. Bioessays. 16(9):633-638.
51.Maher L. J., Wold B., and Dervan P. B. 1989. Inhibition of DNA binding proteins by oligonucleotide-directed triple helix formation. Science. 18; 245(4919): 725-730.
52.Helene C. 1991. Rational design of sequence-specific oncogene inhibitors based on antisense and antigene oligonucleotides. Eur J Cancer. 27(11): 1466-1471.
53.Wang G., Levy D. D., Seidman M. M., and Glazer P. M. 1995. Targeted mutagenesis in mammalian cells mediated by intracellular triple helix formation. Mol Cell Biol. 15(3): 1759-1768.
54.Wang G., Levy D. D., Seidman M. M., and Glazer P. M. 1996. Mutagenesis in mammalian cells induced by triple helix formation and transcription-coupled repair. Science. 9;271(5250):802-805.
55.Wagner R. W. 1994. Gene inhibition using antisense oligodeoxynucleotides. Nature. 24;372(6504):333-335.
56.Kronenwett R., and Haas R. 1998. Antisense strategies for the treatment of hematological malignancies and solid tumors. Ann Hematol. 77(1-2):1-12.
57.Ratajczak M. Z., Kant J. A., Luger S. M., Hijiya N., Zhang J., Zon G., and Gewirtz A. M. 1992. In vivo treatment of human leukemia in a scid mouse model with c-myb antisense oligodeoxynucleotides. Proc. Natl. Acad. Sci. USA. 15;89(24):11823-11827.
58.Alama A., Barbieri F., Cagnoli M., and Schettini G. 1997. Antisense oligonucleotides as therapeutic agents. Pharmacol Res. 36(3):171-178.
59.Eckstein F. 1983. Phosphorothioate analogues of nucleotides-Tools for the investigation of ionchemical processes. Angewandte Chem. 22, 423.
60.Miller P.S., K. B. McParland, K. Jayaramon, and P. O. Ts’o. 1981. Biochemical and biological effects of nonionic nucleic acid methylphosphonates. Biochemistry. 31;20(7):1874-1880.
61.Agrawal S., and R. P. Iyer. 1995. Modified oligonucleotides as therapeutic and diagnostic agents. Curr Opin Biotechnol. 6(1):12-19.
62.Tidd D. M., and Warenius H. M. 1989. Partial protection of oncogene, anti-sense oligodeoxynucleotides against serum nuclease degradation using terminal methylphosphonate groups. Br J Cancer. Sep;60(3):343-350.
63.Forstova J., Krauzewicz N., Wallace S., Street A. J., Dilworth S. M., Beard S., Griffin B. E. 1993. Cooperation of structural proteins during late events in the life cycle of polyomavirus. Journal of Virology. 67:1405-1413.
64.Gillock E. T., Rottinghaus S., Chang D., Cai X., Smiley S. A., Consigli R. A. 1997. Polyomavirus major capsid protein VP1 is capable of packaging cellular DNA when expressed in the baculovirus system. Journal of Virology. 71:2857-2865.
65.Montross L., Watkins S., Moreland R. B., Mamon H., Caspar D. L. D., Garcea R. L. 1991. Nuclear assembly of polyomavirus capsids in insect cells expressing the major capsid protein VP1. Journal of Virology. 65:4991-4998.
66.Sandalon Z., Dalyot-herman N., Oppenhelm A. B., Oppenhelm A. 1997. In vitro assembly of SV40 virions and pseudovirions: Vector development for gene therapy. Human Gene Therapy. 8:843-849.
67.Chang D., Cai X., Consigli R. A. 1993. Characterization of the DNA binding properties of polyomavirus capsid proteins. Journal of Virology. 67:6327-6331.
68.Moreland R. B., Montross L., Garcea R. L. 1991. Characterization of the DNA-binding properties of the polyomavirus capsid protein VP1. Journal of Virology. 65:1168-1176.
69.Soussi T. 1986. DNA-binding properties of the major structural protein of simian virus 40. Journal of Virology. 59:740-744.
70.Clever J., Dean D. A., Kasamatsu H. 1993. Identification of a DNA binding domain in simian virus 40 capsid protein VP2 and VP3. Journal of Biological Chemistry. 268:20877-20883.
71.Pawlita M., Muller M., Oppenlander M., Zentgraf H., Herrman M. 1996. DNA encapsidation by viruslike particles assembled in insect cells from the major capsid protein VP1 of B-lymphotropic papovavirus. Journal of Virology. 70:7517-7526.
72.DeCaprio J. A., Ludlow J. W., Figge J., Shew J. Y., Huang C. M., Lee W. H., Marsilio E., Paucha E., and Livingston D. M. 1988. SV40 large tumor antigen forms a specific complex with the product of the retinoblastoma susceptibility gene. Cell. 15;54(2):275-283.
73.Lin J. Y., Simmons D. T. 1991. The ability of large T antigen to complex with p53 is necessary for the increased life span and partial transformation of human cells by simian virus 40. J Virol. 65(12):6447-6453.
74.Ausubet F. M., Brent R., Kingston R. E., Moore D. D., Seidman J. G., Smith J. A., Struhl K.. (ed) 1988. Current protocols in molecular biology. John Wiley & Sons, Inc., New York.
75.Chang D., Liou Z. M., Ou W. C., Tsai R. T., Wang K. Z., Wang M., Fung C. Y. 1996. Production of the antigen and the antibody of the JC virus major capsid protein VP1. Journal of Virology Methods 59:177-187.
76.Atwood W. J., Wei G., Liu C. K. 1998. Infection of glial cells by the human polyomavirus JC is mediated by N- linked glycoprotein containing terminal α(2,6)- linked sialic acid. Journal of Virology. 72: 4643-4649.

QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top