臺灣博碩士論文加值系統

登革病屬黃質病屬,可藉蚊子傳播,造成人類登革熱、登革出血熱和登革休克症等症狀,為目前世界重大疾病之一。對於很多病患在第二次感染時會引起較為嚴重的出血熱及登革休克症的現象,有人提出了抗體促進效應的假說,即假設登革病毒可能是藉由其表面膜蛋白和吞噬細胞Fc受器結合而進入細胞表面。但是登革病毒如何進行第一次的感染和能感染沒有Fc受器的細胞呢?1997年,陳等人指出登革病毒是藉由細表面的醣胺基進入Vero細胞,1999年,另一陳等人亦報導登革病毒感染單核球需單核球表面分子CD14的幫助。本研究中,我們先加入不同的醣胺基和登革病毒作用,然後觀察病毒感染單核球的能力是否會受到抑制。結果發現只有肝素可以抑制病毒感染單核球,顯示heparan sulfate可能是登革病毒進入細胞內的一個重要因子。另一方面,我們以維生素D3處理沒有表現CD14膜分子的前單核球細胞株U937及THP-1以促使它們分化成類巨噬細胞株而表現膜蛋白CD14後,再以登革病毒感染。結果發現CD14的表現並沒有幫助它們對病毒的感染性。此結果可能是維生素D3的加入活化細胞,因而使細胞對病毒的侵入具有抗性之故。另外,我們以登革病毒感染單核球時,發現它會釋出比較多的細胞介素TNF-alpha。這和在臨床之登革出血熱及登革休克症的病人身上亦發現TNF-alpha分泌失序的情形類似。但,我們進一步探討登革病毒對細胞的影時,卻發現它既不會使細胞活化亦不會造成細胞死亡。至於登革病毒感染單核球後會造成何種生物特性的影響而使第二次感染誘發較嚴重的症狀,仍然需要進一步研究。

Dengue virus is an arthropod-borne human pathogen that represents a serious public health threat. The clinical manifestations of dengue virus infection range in severity from a simple febrile illness to a hemorrhagic fever and a potentially fatal hemorrhagic shock syndrome. Dengue virus gains entry to some cell types by an immune adherence bridging phenomenon which is dependent on the presence of nonneutralizing antibodies. However, this mechanism cannot explain primary infections in patients without antibody to dengue virus and the ability of dengue virus to infect non-phagocytic cell types that do not express Fc receptors. In this study, we treated dengue virus with various GAGs to observe if GAGs can inhibit virus infection. The result indicated that just only heparin could inhibit virus infection. This suggests that heparan sulfate may be an important factor for the penetration of virus inot the host cells. On other hand, we found that the expressing of CD14 couldn’t increase cell’s susceptibility to dengue virus infection. The possible explanation is that the activation of cell by vitamin D3 made cell more resistant to virus infection. Besides, we found that the dengue virus infected cells released more cytokine, TNF-alpha. The phenomenon is similar to those patients with DHF and DSS. To reveal why dengue virus can induce the serious syndrome in the secondary infection to human monocyte need further study.

前言..............1
材料與方法........8
結果..............16
討論..............25
圖................30
參考文獻..........53

Arends, M. J., Morris, R. G. and Wyllie, A. H. 1990. Apoptosis. The role of the endonucleas. Am. J. Pathol. 136:593-608.
Bhamarapravati, N., Tuchinda, P. and Boonyapaknavik, V. 1967. Pathology of Thailand hemorrhagic fever: a study of 100 autopsy cases. Ann. Trop. Med. Parasitol. 61:500-510.
Chen, Y. C., Wang, S. Y. and King, C. C. 1999. Bacterial lipopolysaccharide inhibits dengue virus infection of primary human monocyte/macrophages by blockade of virus entry via a CD14-dependent mechanism. J. Virol. 73:2650-2657.
Chen, Y., Maguire, T., Hileman, R. E., Fromm, J. R., Esko, J. D., Linhardt, R. J. and Marks, R. M. 1997. Dengue virus infectivity depends on envelope protein binding to target cell heparan sulfate. Nat. Med. 3:866-871.
Coligan, J. E., Kruisbeek, A. M., Margulies, D. H., Shevach, E. M. and Strober, W. 1994. In: Current Protocols in Immunology, Vol. 2, Section 7, Immunological Studies in Humans. John Wiley & Sons, USA, pp. 7.6.1-7.6.8.
Columbano, A. 1995. Cell death: current difficulties in discriminating apoptosis from necrosis in the context of pathological processes in vivo. J. Cell. Biochem. 58: 181-190.
Daughaday, C. C., Brandt, W. E., McCown, J. M. and Russell, P. K. 1981. Evidence for two mechanisms of dengue virus infection of adherent human monocytes: trypsin-sensitive virus receptors and trypsin-resistant immune complex receptors. Infect. Immun. 32:469-473.
Defacque, H., Piquemal, D., Basset, A., Marti, J. and Commes, T. 1999. Transforming growth factor-beta 1 is an autocrine mediator of U937 cell growth arrest and differentiation induced by vitamin D3 and retinoids. J. Cell. Physiol. 178: 109-119.
Ellis, R. E., Yuan, J. and Horvitz, H. R. 1991. Mechanisms and functions of cell death. Annu. Rev. Cell. Biol. 7:663-698.
Field, B. N., Kinpe, D. M., Howley, P. M., Chanock, R. M., Melnick, J. L., Monath, T. P., Roizman, B. and Straus, S. E. 1996. Fields Virology. 3rd edition volume 1: 931-1034.
Folkman, J. and Moscona, A. 1978. Role of cell shape in growth control. Nature. 273: 345-349.
Graziani-Bowering, G. M., Graham, J. M. and Filion, L. G. 1997. A quick, easy and inexpensive method for the isolation of human peripheral blood monocytes. J. Immuno. Method. 207:157-168.
Gubler, D. J. 1994. In encyclopedia of virology, pp.3234-331. Edited by Webster, R. G. and Granoff, A. Academic Press, Inc., San Diego, Califormia.
Halstead, S. B. 1988. Pathogenesis of dengue: challenges to molecular biology. Science. 239:476-481.
Halstead, S. B. 1989. Antibody, macrophage, dengue virus infection, shock and hemorrhage: a pathogenetic cascade. Rev. Infect. Dis. 11:S830-S839.
Imbert, J. L., Guevara, P., Ramos, C. J., Ramos, C. and Sotelo, J. 1994. Dengue virus infects mouse cultured neuron but not astrocyte. J. Med. Virol. 42: 228-233.
Jackson, T., Ellard, F. M., Ghazaleh, R. A., Brookes, S. M., Blakemore, W. E., Corteyn, A. H., Stuart, D. I., Newman, W. I. and King, A. Q. 1996. Efficient infection of cells in culture by type O foot-and-mouth disease virus requires binding to cell surface heparan sulfate. J. Virol. 70:5282-5287.
Kaplan, M., Williams, K. J., Mandel, H. and Aviram, M. Role of macrophage glycosaminoglycans in the cellular catabolism of oxidized LDL by macrophages. Arteriosclerosis, Thromb. Vas. Biol. 18:542-553.
Lanciotti, R. S., Calisher, C. H., Gubler, D. J., Chang, G. J. and Vorndam, A. V. 1992. Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase polymerase chain reaction. J. Clin. Micro. Biol. 30:545-551.
McGee, M. P., Teuschler, H. Parthasarathy, N. and Wagner, W. D. 1995. Specific regulation of procoagulant activity on monocytes. Intrinsic pathway inhibition by chondroitin 4,6,-disulfate. J. Biol. Chem. 270:26109-26115.
Monath, T. P. 1994. Dengue; The risk to developed and developing countries. Proc. Natl. Acad. Sci. USA. 91:2395-2400.
Monath, T. P. and Heinzl, F. X. 1996. Flaviviruses. P.961-1034. In B. N. Fields, D. M. Knipe, P. M. Howley, et al.(ed.), Fields Virology. Lippincott-Raven Publisher, Philadelphia.
Moran, L. A., Scrimgeour, K. G., Horton, H. R., Ochs, R. S. and Rawn, J. D. 1994. Biochemistry. 2nd edition volume 9:39-45.
Obunike, J. C., Paka, S., Pillarisetti, S. and Goldberg, I. J. 1997. Lipoprotein lipase can function as a monocyte adhesion protein. Arteriosclerosis, Thromb. Vas. Biol. 17:1414-1420.
Raff, M. C. 1992. Social controls on cell survival and cell death. Nature (London). 356: 397-400.
Rodriguez-Tan, R. S. and Weir, M. R. 1998. Dengue: a view. [Revies]. Texas. Med. 94:53-9.
Russell, P. K. and Nisalak, A. 1967. Dengue virus identification by the plaque reduction neutralization test. J. Immunol. 99:291-296.
Sangkawhibha, N., Rohanasuphot, S. and Ahandrik, S. 1984. Risk factors in dengue shock syndrome: A prospective study in Rayong, thailand. in the 1980 outbreak. Am. J. Epidemiol. 120:653-669.
Shieh, M. T., WuDunn, D., Montgomery, I., Esko, J. D. and Spear, P. G. 1992. Cell surface receptors for herpes simplex virus are heparan sulfate proteoglycans. J. Cell. Biol. 116:1273-1281.
Spear, P. G. 1993. Entry of alphaherpesviruses into cells. Semin. Virol. 4:167-180.
Spear, P. G., Shieh, M. T., Herold, B. C., WuDunn, D. and Koshy, T. I. 1992. Heparan sulfate glycosaminoglycans as primary cell surface receptor for herpes simplex virus. Adv. Exp. Med. Biol. 313:341-353.
Testa, U., Masciulli, R., Tritarellli, E., Pustorino, R., Mariani, G., Martucci, R., Barberi, T., Camagna, A., Valtieri, M. and Peschle, C. 1993. Transfroming growth factor-beta potentiates vitamin D3-induced terminal monocytic differentiation of human leukemic cell lines. J. Immunol. 150:2418-2430.
Vijayagopal, P., Figueroa, J. E., Fontenot, J. D. and Glancy, D. L. 1996. Isolation and characterization of a proteoglycan variant from human aorta exhibiting a marked affinity for low density lipoprotein and demonstration of its enhanced expression in atherosclerotic plaques. Atherosclerosis. 127:195-203.
Wyllie, A. H., Kerr, J. F. and Curries, A. R. 1980. Cell death: the significance of apoptosis. Inter. Rev. Cytol. 68:251-306.
Yadav, M., Kamath, K. R. Iyngkaran, N. and Sinniah, M. 1991. Dengue haemorrhagic fever and dengue shock syndrome: are they tumour necrosis factor-mediated disorders? FEMS Micro. Immuno. 4:45-49.
Yeaman, C. and Rapraeger, A. C. 1993. Membrane-anchored proteoglycans of mouse macrophages: P388D1 cells express a syndecan-4 lile heparan sulfate proteoglycan and a distinct chondroitin sulfate form. J. Cell. Physiol. 157:413-425.