一直以來，感染C型肝炎病毒是造成慢性肝炎和肝癌的主要兇手。過去研究顯示，它可以藉由和許多細胞表面受體的結合作用來感染人類的肝臟細胞，其中包括了CD81,SR-BI,claudin-1和occludin。也有不少報告指出，凝集素受體可以辨識C型肝炎病毒並間接地幫助它進入細胞中，而它的外套膜蛋白E1和E2是具有高度醣化的醣蛋白，這個特點更加指明了凝集素受體和病毒之間的結合作用在C型肝炎病毒感染細胞時的重要性。然而，其外套膜蛋白上醣化作用的多樣性是否可改變與這些細胞表面受體和凝集素受體的親和力，這方面的研究卻是極為有限。在目前的研究中，我們利用表面薄膜共振生物感應器來分析帶有不同醣化形式外套膜蛋白與各個受體的親和力，並試著生產帶有不同醣化形式的偽病毒(pseudovirus)來檢測它對病毒的組裝和感染力的影響。我們的研究結果顯示，在鈣離子存在的環境中，凝集素類的受體(DC-SIGN, L-SIGN和Langerin)相較於非凝集素類受體，與病毒的外套膜蛋白具有相當高的親和力，差距最大可達至萬倍。而其中高甘露糖型(high-mannose type)的外套膜蛋白，甘露糖的組成越高親和力就越好，Man8/9形式的外套膜蛋白與凝集素類受體的親和力可以高過Man5型和complex type約十倍。令人感興趣的是，帶有Man5形式的外套膜蛋白和非凝集素類的受體其親和力在這三種醣化形式中是最好的。而在進一步的實驗中，我們發現不同醣化形式的偽病毒，組裝病毒的能力和感染力的確都有受到影響。帶有Man5形式外套膜蛋白的偽病毒其感染Huh7細胞株的能力是最強的，而帶有Man8/9形式外套膜蛋白的偽病毒其在細胞中的蛋白質表現量和組裝蛋白質至病毒體上的量都是最高的。總結我們的研究結果，可以發現C型肝炎病毒外套膜蛋白上的醣類組成能影響病毒的組裝能力，也能藉由和各種細胞上能辨識此蛋白質的受體來調節其感染力。這些具有與病毒外套膜蛋白高度親和力的受體，也許可以為C型肝炎病毒的治療策略提供一個新的方向。

Hepatitis C virus (HCV) infection is a major cause of chronic hepatitis and hepatocellular carcinoma worldwide. It infects human liver cells through interacting with several cellular protein receptors including CD81, SR-BI, Claudin-1 and occludin. There are accumulating reports indicating that lectin receptors can mediate HCV recognition and entry in the body. The envelope proteins of HCV (E1 and E2) are heavily glycosylated, further address the role of lectin receptor-viral particle interaction in HCV infection. However, there is limited report investigating the relation of glycoforms of HCV envelope proteins with the affinity toward lectin and non-lectin receptors. Here we used surface plasmon resonance (SPR) to examine the binding affinity of different glycoforms of HCV envelope protein to receptors, and inspected the infectivity and assembly of HCV pseudoparticles composed with different glycoforms of envelope proteins. Our results indicated that DC-SIGN, L-SIGN and Langerin had higher affinity to bind HCV envelope proteins in the presence of calcium ions than non-lectin receptors (SR-BI, CD81, claudin-1, and occludin), and envelope proteins with Man8/9 N-glycans showed approximate 10-folds better in binding to lectin receptors than envelopes proteins with Man5 and complex type N-glycans. Interestingly, comparing among glycoforms, envelope proteins with Man5 N-glycans showed the highest binding affinity when interacting with non-lectin receptors. It was also revealed that HCV pseudoparticles with Man5 N-glycans, among the glycoforms we tested, carried the best infectivity toward hepatoma cell lines. In summary, glycans on HCV envelope protein play a modulatory role in HCV assembly and infection, and direct HCV-receptor interaction, which mediates viral entry in different cells. Receptors with high affinity in binding to HCV envelope proteins may help establish therapeutic strategy toward HCV.

摘要 ii
Abstract iii
Table of Contents v
Table of Contents/Figures ix
Table of Contents/Tables xi
1. INTRODUCTIONS 1
1.1 An overview of Hepatitis C virus (HCV) 2
1.1.1 Discovery of HCV 2
1.1.2 HCV Transmission and Epidemiology 2
1.1.3 HCV Pathogenesis 3
1.1.4 Current Treatment 4
1.2 HCV Structure, Genome Organization and Protein Function 5
1.2.1 HCV particle 5
1.2.2 Genome Organization 6
1.2.3 Structural proteins of HCV 7
1.2.4 Non-structural proteins of HCV 8
1.3 HCV Lifecycle 10
1.4 Cellular entry factors for HCV 11
1.4.1 Attachment factors 11
1.4.2 Receptors 14
1.5 General Principles of Glycosylation 17
1.5.1. N-glycan biosynthesis 18
1.5.2. Glycoenzymes involved in N-glycan biosynthesis 19
1.6 Glycosylation on HCV 20
1.7 Significance and Purpose 22
2. MATERIALS AND METHODS 29
2.1 Materials 30
2.2 Methods 34
2.2.1 Cell culture 34
2.2.2 Plasmid construction 34
2.2.3 Cell transfection 37
2.2.4 Purification of FLAG-tagged full-length E2 proteins, Fc-tagged lectins, Fc-tagged HCV protein receptors, and His-tagged soluble E1E2 proteins 38
2.2.5 Western blotting 40
2.2.6 Enzyme-linked immunosorbent assay (ELISA) 41
2.2.7 In Solution Tryptic-Chymotryptic Digestion, N-Glycosite Determination and Assigning Glycopeptides 42
2.2.8 Surface plasmon resonance (SPR) analysis 43
2.2.9 Pseudovirus generation and infection assays 45
2.2.10 Glycosidase treatment 47
2.2.11 Lectins blocking assay for HCVpp infection 48
3. RESULTS 49
3.1 Interaction between HCV E2 protein and Fc-Innate Immune receptors 50
3.2 Characterization of the N-linked glycosylation of full-length hepatitis C virus E2 envelope glycoprotein 51
3.3 Characterization of N-linked glycosylation of truncated hepatitis C virus E1 and E2 envelope glycoprotein 52
3.4 Binding affinities of different HCV E1/E2 glycoforms toward receptors 54
3.5 Langerin and L-SIGN in blocking HCVpp infection 58
3.6 Assembly and infectivity of different glycoforms of HCVpp 59
3.7 Glycoforms of E2 proteins from HCVpp 61
4. DISCUSSION 63
4.1 Protein structure, glycosylation and cellular localization 64
4.2 Glycoforms of HCV envelope proteins 66
4.3 Receptors for HCV 69
4.3.1 Lectin-like receptors for HCV 69
4.3.2 Non-lectin receptors for HCV 73
4.3.3 Roles of lectin-like receptors for HCV binding, infection, and propagation 74
4.4 Glycoforms of envelope proteins influence HCVpp assembly and infectivity 76
4.5 Targeting HCV envelope protein 77
4.6 Prospects 77
5. FIGURES 79
6. TABLES 109
7. REFERENCES 113

Acton, S., A. Rigotti, K. T. Landschulz, S. Xu, H. H. Hobbs, and M. Krieger. 1996. Identification of scavenger receptor SR-BI as a high density lipoprotein receptor. Science 271:518-520.
Acton, S. L., P. E. Scherer, H. F. Lodish, and M. Krieger. 1994. Expression cloning of SR-BI, a CD36-related class B scavenger receptor. The Journal of biological chemistry 269:21003-21009.
Agnello, V., G. Abel, M. Elfahal, G. B. Knight, and Q. X. Zhang. 1999. Hepatitis C virus and other flaviviridae viruses enter cells via low density lipoprotein receptor. Proceedings of the National Academy of Sciences of the United States of America 96:12766-12771.
Alter, H. J., P. V. Holland, A. G. Morrow, R. H. Purcell, S. M. Feinstone, and Y. Moritsugu. 1975. Clinical and serological analysis of transfusion-associated hepatitis. Lancet 2:838-841.
Alter, M. J. 2007. Epidemiology of hepatitis C virus infection. World journal of gastroenterology 13:2436-2441.
Ambudkar, S. V., C. Kimchi-Sarfaty, Z. E. Sauna, and M. M. Gottesman. 2003. P-glycoprotein: from genomics to mechanism. Oncogene 22:7468-7485.
Andre, P., F. Komurian-Pradel, S. Deforges, M. Perret, J. L. Berland, M. Sodoyer, S. Pol, C. Brechot, G. Paranhos-Baccala, and V. Lotteau. 2002. Characterization of low- and very-low-density hepatitis C virus RNA-containing particles. Journal of virology 76:6919-6928.
Ashfaq, U. A., M. S. Masoud, S. Khaliq, Z. Nawaz, and S. Riazuddin. 2011. Inhibition of hepatitis C virus 3a genotype entry through Glanthus Nivalis Agglutinin. Virology journal 8:248.
Auffermann-Gretzinger, S., E. B. Keeffe, and S. Levy. 2001. Impaired dendritic cell maturation in patients with chronic, but not resolved, hepatitis C virus infection. Blood 97:3171-3176.
Bain, C., A. Fatmi, F. Zoulim, J. P. Zarski, C. Trepo, and G. Inchauspe. 2001. Impaired allostimulatory function of dendritic cells in chronic hepatitis C infection. Gastroenterology 120:512-524.
Barth, H., C. Schafer, M. I. Adah, F. Zhang, R. J. Linhardt, H. Toyoda, A. Kinoshita-Toyoda, T. Toida, T. H. Van Kuppevelt, E. Depla, F. Von Weizsacker, H. E. Blum, and T. F. Baumert. 2003. Cellular binding of hepatitis C virus envelope glycoprotein E2 requires cell surface heparan sulfate. The Journal of biological chemistry 278:41003-41012.
Barth, H., A. Ulsenheimer, G. R. Pape, H. M. Diepolder, M. Hoffmann, C. Neumann-Haefelin, R. Thimme, P. Henneke, R. Klein, G. Paranhos-Baccala, E. Depla, T. J. Liang, H. E. Blum, and T. F. Baumert. 2005. Uptake and presentation of hepatitis C virus-like particles by human dendritic cells. Blood 105:3605-3614.
Bartosch, B., A. Vitelli, C. Granier, C. Goujon, J. Dubuisson, S. Pascale, E. Scarselli, R. Cortese, A. Nicosia, and F. L. Cosset. 2003. Cell entry of hepatitis C virus requires a set of co-receptors that include the CD81 tetraspanin and the SR-B1 scavenger receptor. The Journal of biological chemistry 278:41624-41630.
Basu, A., T. Kanda, A. Beyene, K. Saito, K. Meyer, and R. Ray. 2007. Sulfated homologues of heparin inhibit hepatitis C virus entry into mammalian cells. Journal of virology 81:3933-3941.
Beasley, R. P., L. Y. Hwang, C. C. Lin, and C. S. Chien. 1981. Hepatocellular carcinoma and hepatitis B virus. A prospective study of 22 707 men in Taiwan. Lancet 2:1129-1133.
Bekisz, J., H. Schmeisser, J. Hernandez, N. D. Goldman, and K. C. Zoon. 2004. Human interferons alpha, beta and omega. Growth Factors 22:243-251.
Boulant, S., C. Vanbelle, C. Ebel, F. Penin, and J. P. Lavergne. 2005. Hepatitis C virus core protein is a dimeric alpha-helical protein exhibiting membrane protein features. Journal of virology 79:11353-11365.
Bradley, D., K. McCaustland, K. Krawczynski, J. Spelbring, C. Humphrey, and E. H. Cook. 1991. Hepatitis C virus: buoyant density of the factor VIII-derived isolate in sucrose. Journal of medical virology 34:206-208.
Brautigam, J., A. J. Scheidig, and W. Egge-Jacobsen. 2013. Mass spectrometric analysis of hepatitis C viral envelope protein E2 reveals extended microheterogeneity of mucin-type O-linked glycosylation. Glycobiology 23:453-474.
Brazzoli, M., A. Bianchi, S. Filippini, A. Weiner, Q. Zhu, M. Pizza, and S. Crotta. 2008. CD81 is a central regulator of cellular events required for hepatitis C virus infection of human hepatocytes. Journal of virology 82:8316-8329.
Brown, K. S., M. J. Keogh, A. M. Owsianka, R. Adair, A. H. Patel, J. N. Arnold, J. K. Ball, R. B. Sim, A. W. Tarr, and T. P. Hickling. 2010. Specific interaction of hepatitis C virus glycoproteins with mannan binding lectin inhibits virus entry. Protein & cell 1:664-674.
Bukh, J., R. H. Purcell, and R. H. Miller. 1994. Sequence analysis of the core gene of 14 hepatitis C virus genotypes. Proceedings of the National Academy of Sciences of the United States of America 91:8239-8243.
Burlone, M. E., and A. Budkowska. 2009. Hepatitis C virus cell entry: role of lipoproteins and cellular receptors. The Journal of general virology 90:1055-1070.
Callens, N., Y. Ciczora, B. Bartosch, N. Vu-Dac, F. L. Cosset, J. M. Pawlotsky, F. Penin, and J. Dubuisson. 2005. Basic residues in hypervariable region 1 of hepatitis C virus envelope glycoprotein e2 contribute to virus entry. Journal of virology 79:15331-15341.
Catanese, M. T., H. Ansuini, R. Graziani, T. Huby, M. Moreau, J. K. Ball, G. Paonessa, C. M. Rice, R. Cortese, A. Vitelli, and A. Nicosia. 2010. Role of scavenger receptor class B type I in hepatitis C virus entry: kinetics and molecular determinants. Journal of virology 84:34-43.
Catanese, M. T., R. Graziani, T. von Hahn, M. Moreau, T. Huby, G. Paonessa, C. Santini, A. Luzzago, C. M. Rice, R. Cortese, A. Vitelli, and A. Nicosia. 2007. High-avidity monoclonal antibodies against the human scavenger class B type I receptor efficiently block hepatitis C virus infection in the presence of high-density lipoprotein. Journal of virology 81:8063-8071.
Chang, K. S., J. Jiang, Z. Cai, and G. Luo. 2007. Human apolipoprotein e is required for infectivity and production of hepatitis C virus in cell culture. Journal of virology 81:13783-13793.
Chen, Y., T. Maguire, R. E. Hileman, J. R. Fromm, J. D. Esko, R. J. Linhardt, and R. M. Marks. 1997. Dengue virus infectivity depends on envelope protein binding to target cell heparan sulfate. Nature medicine 3:866-871.
Choo, Q. L., G. Kuo, A. J. Weiner, L. R. Overby, D. W. Bradley, and M. Houghton. 1989. Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 244:359-362.
Choukhi, A., S. Ung, C. Wychowski, and J. Dubuisson. 1998. Involvement of endoplasmic reticulum chaperones in the folding of hepatitis C virus glycoproteins. Journal of virology 72:3851-3858.
Cocquerel, L., S. Duvet, J. C. Meunier, A. Pillez, R. Cacan, C. Wychowski, and J. Dubuisson. 1999. The transmembrane domain of hepatitis C virus glycoprotein E1 is a signal for static retention in the endoplasmic reticulum. Journal of virology 73:2641-2649.
Cocquerel, L., C. C. Kuo, J. Dubuisson, and S. Levy. 2003. CD81-dependent binding of hepatitis C virus E1E2 heterodimers. Journal of virology 77:10677-10683.
Cocquerel, L., A. Op de Beeck, M. Lambot, J. Roussel, D. Delgrange, A. Pillez, C. Wychowski, F. Penin, and J. Dubuisson. 2002. Topological changes in the transmembrane domains of hepatitis C virus envelope glycoproteins. The EMBO journal 21:2893-2902.
Cormier, E. G., R. J. Durso, F. Tsamis, L. Boussemart, C. Manix, W. C. Olson, J. P. Gardner, and T. Dragic. 2004a. L-SIGN (CD209L) and DC-SIGN (CD209) mediate transinfection of liver cells by hepatitis C virus. Proceedings of the National Academy of Sciences of the United States of America 101:14067-14072.
Cormier, E. G., F. Tsamis, F. Kajumo, R. J. Durso, J. P. Gardner, and T. Dragic. 2004b. CD81 is an entry coreceptor for hepatitis C virus. Proceedings of the National Academy of Sciences of the United States of America 101:7270-7274.
de Witte, L., A. Nabatov, M. Pion, D. Fluitsma, M. A. de Jong, T. de Gruijl, V. Piguet, Y. van Kooyk, and T. B. Geijtenbeek. 2007. Langerin is a natural barrier to HIV-1 transmission by Langerhans cells. Nature medicine 13:367-371.
Deleersnyder, V., A. Pillez, C. Wychowski, K. Blight, J. Xu, Y. S. Hahn, C. M. Rice, and J. Dubuisson. 1997. Formation of native hepatitis C virus glycoprotein complexes. Journal of virology 71:697-704.
Drummer, H. E., I. Boo, A. L. Maerz, and P. Poumbourios. 2006. A conserved Gly436-Trp-Leu-Ala-Gly-Leu-Phe-Tyr motif in hepatitis C virus glycoprotein E2 is a determinant of CD81 binding and viral entry. Journal of virology 80:7844-7853.
Dubuisson, J. 2007. Hepatitis C virus proteins. World journal of gastroenterology 13:2406-2415.
Dubuisson, J., H. H. Hsu, R. C. Cheung, H. B. Greenberg, D. G. Russell, and C. M. Rice. 1994. Formation and intracellular localization of hepatitis C virus envelope glycoprotein complexes expressed by recombinant vaccinia and Sindbis viruses. Journal of virology 68:6147-6160.
Dubuisson, J., F. Penin, and D. Moradpour. 2002. Interaction of hepatitis C virus proteins with host cell membranes and lipids. Trends in cell biology 12:517-523.
Dubuisson, J., and C. M. Rice. 1996. Hepatitis C virus glycoprotein folding: disulfide bond formation and association with calnexin. Journal of virology 70:778-786.
Duvet, S., L. Cocquerel, A. Pillez, R. Cacan, A. Verbert, D. Moradpour, C. Wychowski, and J. Dubuisson. 1998. Hepatitis C virus glycoprotein complex localization in the endoplasmic reticulum involves a determinant for retention and not retrieval. The Journal of biological chemistry 273:32088-32095.
Egger, D., B. Wolk, R. Gosert, L. Bianchi, H. E. Blum, D. Moradpour, and K. Bienz. 2002. Expression of hepatitis C virus proteins induces distinct membrane alterations including a candidate viral replication complex. Journal of virology 76:5974-5984.
Evans, M. J., C. M. Rice, and S. P. Goff. 2004. Phosphorylation of hepatitis C virus nonstructural protein 5A modulates its protein interactions and viral RNA replication. Proceedings of the National Academy of Sciences of the United States of America 101:13038-13043.
Evans, M. J., T. von Hahn, D. M. Tscherne, A. J. Syder, M. Panis, B. Wolk, T. Hatziioannou, J. A. McKeating, P. D. Bieniasz, and C. M. Rice. 2007. Claudin-1 is a hepatitis C virus co-receptor required for a late step in entry. Nature 446:801-805.
Falkowska, E., F. Kajumo, E. Garcia, J. Reinus, and T. Dragic. 2007. Hepatitis C virus envelope glycoprotein E2 glycans modulate entry, CD81 binding, and neutralization. Journal of virology 81:8072-8079.
Farquhar, M. J., K. Hu, H. J. Harris, C. Davis, C. L. Brimacombe, S. J. Fletcher, T. F. Baumert, J. Z. Rappoport, P. Balfe, and J. A. McKeating. 2012. Hepatitis C virus induces CD81 and claudin-1 endocytosis. Journal of virology 86:4305-4316.
Feinstone, S. M., A. Z. Kapikian, R. H. Purcell, H. J. Alter, and P. V. Holland. 1975. Transfusion-associated hepatitis not due to viral hepatitis type A or B. The New England journal of medicine 292:767-770.
Feld, J. J., and J. H. Hoofnagle. 2005. Mechanism of action of interferon and ribavirin in treatment of hepatitis C. Nature 436:967-972.
Flint, M., J. Dubuisson, C. Maidens, R. Harrop, G. R. Guile, P. Borrow, and J. A. McKeating. 2000. Functional characterization of intracellular and secreted forms of a truncated hepatitis C virus E2 glycoprotein. Journal of virology 74:702-709.
Flint, M., and J. A. McKeating. 1999. The C-terminal region of the hepatitis C virus E1 glycoprotein confers localization within the endoplasmic reticulum. The Journal of general virology 80 ( Pt 8):1943-1947.
Fukushi, S., M. Okada, T. Kageyama, F. B. Hoshino, K. Nagai, and K. Katayama. 2001. Interaction of poly(rC)-binding protein 2 with the 5''-terminal stem loop of the hepatitis C-virus genome. Virus research 73:67-79.
Gardner, J. P., R. J. Durso, R. R. Arrigale, G. P. Donovan, P. J. Maddon, T. Dragic, and W. C. Olson. 2003. L-SIGN (CD 209L) is a liver-specific capture receptor for hepatitis C virus. Proceedings of the National Academy of Sciences of the United States of America 100:4498-4503.
Germi, R., J. M. Crance, D. Garin, J. Guimet, H. Lortat-Jacob, R. W. Ruigrok, J. P. Zarski, and E. Drouet. 2002a. Cellular glycosaminoglycans and low density lipoprotein receptor are involved in hepatitis C virus adsorption. Journal of medical virology 68:206-215.
Germi, R., J. M. Crance, D. Garin, J. Guimet, H. Lortat-Jacob, R. W. Ruigrok, J. P. Zarski, and E. Drouet. 2002b. Heparan sulfate-mediated binding of infectious dengue virus type 2 and yellow fever virus. Virology 292:162-168.
Goffard, A., N. Callens, B. Bartosch, C. Wychowski, F. L. Cosset, C. Montpellier, and J. Dubuisson. 2005. Role of N-linked glycans in the functions of hepatitis C virus envelope glycoproteins. Journal of virology 79:8400-8409.
Goffard, A., and J. Dubuisson. 2003. Glycosylation of hepatitis C virus envelope proteins. Biochimie 85:295-301.
Grakoui, A., C. Wychowski, C. Lin, S. M. Feinstone, and C. M. Rice. 1993. Expression and identification of hepatitis C virus polyprotein cleavage products. Journal of virology 67:1385-1395.
Griffin, S. D., L. P. Beales, D. S. Clarke, O. Worsfold, S. D. Evans, J. Jaeger, M. P. Harris, and D. J. Rowlands. 2003. The p7 protein of hepatitis C virus forms an ion channel that is blocked by the antiviral drug, Amantadine. FEBS letters 535:34-38.
Hadziyannis, S. J., H. Sette, Jr., T. R. Morgan, V. Balan, M. Diago, P. Marcellin, G. Ramadori, H. Bodenheimer, Jr., D. Bernstein, M. Rizzetto, S. Zeuzem, P. J. Pockros, A. Lin, and A. M. Ackrill. 2004. Peginterferon-alpha2a and ribavirin combination therapy in chronic hepatitis C: a randomized study of treatment duration and ribavirin dose. Annals of internal medicine 140:346-355.
Harris, H. J., C. Davis, J. G. Mullins, K. Hu, M. Goodall, M. J. Farquhar, C. J. Mee, K. McCaffrey, S. Young, H. Drummer, P. Balfe, and J. A. McKeating. 2010. Claudin association with CD81 defines hepatitis C virus entry. The Journal of biological chemistry 285:21092-21102.
Harris, H. J., M. J. Farquhar, C. J. Mee, C. Davis, G. M. Reynolds, A. Jennings, K. Hu, F. Yuan, H. Deng, S. G. Hubscher, J. H. Han, P. Balfe, and J. A. McKeating. 2008. CD81 and claudin 1 coreceptor association: role in hepatitis C virus entry. Journal of virology 82:5007-5020.
Helle, F., and J. Dubuisson. 2008. Hepatitis C virus entry into host cells. Cellular and molecular life sciences : CMLS 65:100-112.
Helle, F., A. Goffard, V. Morel, G. Duverlie, J. McKeating, Z. Y. Keck, S. Foung, F. Penin, J. Dubuisson, and C. Voisset. 2007. The neutralizing activity of anti-hepatitis C virus antibodies is modulated by specific glycans on the E2 envelope protein. Journal of virology 81:8101-8111.
Helle, F., G. Vieyres, L. Elkrief, C. I. Popescu, C. Wychowski, V. Descamps, S. Castelain, P. Roingeard, G. Duverlie, and J. Dubuisson. 2010. Role of N-linked glycans in the functions of hepatitis C virus envelope proteins incorporated into infectious virions. Journal of virology 84:11905-11915.
Helle, F., C. Wychowski, N. Vu-Dac, K. R. Gustafson, C. Voisset, and J. Dubuisson. 2006. Cyanovirin-N inhibits hepatitis C virus entry by binding to envelope protein glycans. The Journal of biological chemistry 281:25177-25183.
Hilgard, P., and R. Stockert. 2000. Heparan sulfate proteoglycans initiate dengue virus infection of hepatocytes. Hepatology 32:1069-1077.
Huang, L., J. Hwang, S. D. Sharma, M. R. Hargittai, Y. Chen, J. J. Arnold, K. D. Raney, and C. E. Cameron. 2005. Hepatitis C virus nonstructural protein 5A (NS5A) is an RNA-binding protein. The Journal of biological chemistry 280:36417-36428.
Huang, Z. H., D. Gu, Y. Lange, and T. Mazzone. 2003. Expression of scavenger receptor BI facilitates sterol movement between the plasma membrane and the endoplasmic reticulum in macrophages. Biochemistry 42:3949-3955.
Hugle, T., F. Fehrmann, E. Bieck, M. Kohara, H. G. Krausslich, C. M. Rice, H. E. Blum, and D. Moradpour. 2001. The hepatitis C virus nonstructural protein 4B is an integral endoplasmic reticulum membrane protein. Virology 284:70-81.
Iacob, R. E., I. Perdivara, M. Przybylski, and K. B. Tomer. 2008. Mass spectrometric characterization of glycosylation of hepatitis C virus E2 envelope glycoprotein reveals extended microheterogeneity of N-glycans. Journal of the American Society for Mass Spectrometry 19:428-444.
Ishida, S., M. Kaito, M. Kohara, K. Tsukiyama-Kohora, N. Fujita, J. Ikoma, Y. Adachi, and S. Watanabe. 2001. Hepatitis C virus core particle detected by immunoelectron microscopy and optical rotation technique. Hepatology research : the official journal of the Japan Society of Hepatology 20:335-347.
Jaeckel, E., M. Cornberg, H. Wedemeyer, T. Santantonio, J. Mayer, M. Zankel, G. Pastore, M. Dietrich, C. Trautwein, and M. P. Manns. 2001. Treatment of acute hepatitis C with interferon alfa-2b. The New England journal of medicine 345:1452-1457.
Jirasko, V., R. Montserret, N. Appel, A. Janvier, L. Eustachi, C. Brohm, E. Steinmann, T. Pietschmann, F. Penin, and R. Bartenschlager. 2008. Structural and functional characterization of nonstructural protein 2 for its role in hepatitis C virus assembly. The Journal of biological chemistry 283:28546-28562.
Jones, D. M., A. H. Patel, P. Targett-Adams, and J. McLauchlan. 2009. The hepatitis C virus NS4B protein can trans-complement viral RNA replication and modulates production of infectious virus. Journal of virology 83:2163-2177.
Kaito, M., S. Watanabe, K. Tsukiyama-Kohara, K. Yamaguchi, Y. Kobayashi, M. Konishi, M. Yokoi, S. Ishida, S. Suzuki, and M. Kohara. 1994. Hepatitis C virus particle detected by immunoelectron microscopic study. The Journal of general virology 75 ( Pt 7):1755-1760.
Kawamura, T., S. E. Kurtz, A. Blauvelt, and S. Shimada. 2005. The role of Langerhans cells in the sexual transmission of HIV. Journal of dermatological science 40:147-155.
Kobayashi, F., S. Yamada, S. Taguwa, C. Kataoka, S. Naito, Y. Hama, H. Tani, Y. Matsuura, and K. Sugahara. 2012. Specific interaction of the envelope glycoproteins E1 and E2 with liver heparan sulfate involved in the tissue tropismatic infection by hepatitis C virus. Glycoconjugate journal 29:211-220.
Koutsoudakis, G., A. Kaul, E. Steinmann, S. Kallis, V. Lohmann, T. Pietschmann, and R. Bartenschlager. 2006. Characterization of the early steps of hepatitis C virus infection by using luciferase reporter viruses. Journal of virology 80:5308-5320.
Krieger, S. E., M. B. Zeisel, C. Davis, C. Thumann, H. J. Harris, E. K. Schnober, C. Mee, E. Soulier, C. Royer, M. Lambotin, F. Grunert, V. L. Dao Thi, M. Dreux, F. L. Cosset, J. A. McKeating, C. Schuster, and T. F. Baumert. 2010. Inhibition of hepatitis C virus infection by anti-claudin-1 antibodies is mediated by neutralization of E2-CD81-claudin-1 associations. Hepatology 51:1144-1157.
Lai, W. K., P. J. Sun, J. Zhang, A. Jennings, P. F. Lalor, S. Hubscher, J. A. McKeating, and D. H. Adams. 2006. Expression of DC-SIGN and DC-SIGNR on human sinusoidal endothelium: a role for capturing hepatitis C virus particles. The American journal of pathology 169:200-208.
Lau, J. Y., G. L. Davis, L. E. Prescott, G. Maertens, K. L. Lindsay, K. Qian, M. Mizokami, and P. Simmonds. 1996. Distribution of hepatitis C virus genotypes determined by line probe assay in patients with chronic hepatitis C seen at tertiary referral centers in the United States. Hepatitis Interventional Therapy Group. Annals of internal medicine 124:868-876.
Li, Y., B. Hao, X. Kuai, G. Xing, J. Yang, J. Chen, L. Tang, L. Zhang, and F. He. 2009. C-type lectin LSECtin interacts with DC-SIGNR and is involved in hepatitis C virus binding. Molecular and cellular biochemistry 327:183-190.
Lindenbach, B. D., and C. M. Rice. 2005. Unravelling hepatitis C virus replication from genome to function. Nature 436:933-938.
Liu, R., M. Tewari, R. Kong, R. Zhang, P. Ingravallo, and R. Ralston. 2010a. A peptide derived from hepatitis C virus E2 envelope protein inhibits a post-binding step in HCV entry. Antiviral research 86:172-179.
Liu, S., W. Kuo, W. Yang, W. Liu, G. A. Gibson, K. Dorko, S. C. Watkins, S. C. Strom, and T. Wang. 2010b. The second extracellular loop dictates Occludin-mediated HCV entry. Virology 407:160-170.
Liu, S., W. Yang, L. Shen, J. R. Turner, C. B. Coyne, and T. Wang. 2009. Tight junction proteins claudin-1 and occludin control hepatitis C virus entry and are downregulated during infection to prevent superinfection. Journal of virology 83:2011-2014.
Liu, W., L. Tang, G. Zhang, H. Wei, Y. Cui, L. Guo, Z. Gou, X. Chen, D. Jiang, Y. Zhu, G. Kang, and F. He. 2004. Characterization of a novel C-type lectin-like gene, LSECtin: demonstration of carbohydrate binding and expression in sinusoidal endothelial cells of liver and lymph node. The Journal of biological chemistry 279:18748-18758.
Lohmann, V., A. Roos, F. Korner, J. O. Koch, and R. Bartenschlager. 2000. Biochemical and structural analysis of the NS5B RNA-dependent RNA polymerase of the hepatitis C virus. Journal of viral hepatitis 7:167-174.
Lozach, P. Y., A. Amara, B. Bartosch, J. L. Virelizier, F. Arenzana-Seisdedos, F. L. Cosset, and R. Altmeyer. 2004. C-type lectins L-SIGN and DC-SIGN capture and transmit infectious hepatitis C virus pseudotype particles. The Journal of biological chemistry 279:32035-32045.
Lozach, P. Y., H. Lortat-Jacob, A. de Lacroix de Lavalette, I. Staropoli, S. Foung, A. Amara, C. Houles, F. Fieschi, O. Schwartz, J. L. Virelizier, F. Arenzana-Seisdedos, and R. Altmeyer. 2003. DC-SIGN and L-SIGN are high affinity binding receptors for hepatitis C virus glycoprotein E2. The Journal of biological chemistry 278:20358-20366.
Macdonald, A., and M. Harris. 2004. Hepatitis C virus NS5A: tales of a promiscuous protein. The Journal of general virology 85:2485-2502.
Maillard, P., T. Huby, U. Andreo, M. Moreau, J. Chapman, and A. Budkowska. 2006. The interaction of natural hepatitis C virus with human scavenger receptor SR-BI/Cla1 is mediated by ApoB-containing lipoproteins. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 20:735-737.
Mandl, C. W., H. Kroschewski, S. L. Allison, R. Kofler, H. Holzmann, T. Meixner, and F. X. Heinz. 2001. Adaptation of tick-borne encephalitis virus to BHK-21 cells results in the formation of multiple heparan sulfate binding sites in the envelope protein and attenuation in vivo. Journal of virology 75:5627-5637.
Manns, M. P., J. G. McHutchison, S. C. Gordon, V. K. Rustgi, M. Shiffman, R. Reindollar, Z. D. Goodman, K. Koury, M. Ling, and J. K. Albrecht. 2001. Peginterferon alfa-2b plus ribavirin compared with interferon alfa-2b plus ribavirin for initial treatment of chronic hepatitis C: a randomised trial. Lancet 358:958-965.
Mason, P. W. 1989. Maturation of Japanese encephalitis virus glycoproteins produced by infected mammalian and mosquito cells. Virology 169:354-364.
Matsuura, Y., S. Harada, R. Suzuki, Y. Watanabe, Y. Inoue, I. Saito, and T. Miyamura. 1992. Expression of processed envelope protein of hepatitis C virus in mammalian and insect cells. Journal of virology 66:1425-1431.
Matsuura, Y., T. Suzuki, R. Suzuki, M. Sato, H. Aizaki, I. Saito, and T. Miyamura. 1994. Processing of E1 and E2 glycoproteins of hepatitis C virus expressed in mammalian and insect cells. Virology 205:141-150.
McLauchlan, J. 2000. Properties of the hepatitis C virus core protein: a structural protein that modulates cellular processes. Journal of viral hepatitis 7:2-14.
Merola, M., M. Brazzoli, F. Cocchiarella, J. M. Heile, A. Helenius, A. J. Weiner, M. Houghton, and S. Abrignani. 2001. Folding of hepatitis C virus E1 glycoprotein in a cell-free system. Journal of virology 75:11205-11217.
Michalak, J. P., C. Wychowski, A. Choukhi, J. C. Meunier, S. Ung, C. M. Rice, and J. Dubuisson. 1997. Characterization of truncated forms of hepatitis C virus glycoproteins. The Journal of general virology 78 ( Pt 9):2299-2306.
Michta, M. L., S. E. Hopcraft, C. M. Narbus, Z. Kratovac, B. Israelow, M. Sourisseau, and M. J. Evans. 2010. Species-specific regions of occludin required by hepatitis C virus for cell entry. Journal of virology 84:11696-11708.
Molina, S., V. Castet, C. Fournier-Wirth, L. Pichard-Garcia, R. Avner, D. Harats, J. Roitelman, R. Barbaras, P. Graber, P. Ghersa, M. Smolarsky, A. Funaro, F. Malavasi, D. Larrey, J. Coste, J. M. Fabre, A. Sa-Cunha, and P. Maurel. 2007. The low-density lipoprotein receptor plays a role in the infection of primary human hepatocytes by hepatitis C virus. Journal of hepatology 46:411-419.
Nakayama, Y., N. Nakamura, D. Tsuji, K. Itoh, and A. Kurosaka. 2013. Diseases Associated with Protein Glycosylation Disorders in Mammals. Genetic Disorders, Prof. Maria Puiu (Ed.), ISBN: 978-953-51-0886-3, InTech.
Nowak, T., P. M. Farber, and G. Wengler. 1989. Analyses of the terminal sequences of West Nile virus structural proteins and of the in vitro translation of these proteins allow the proposal of a complete scheme of the proteolytic cleavages involved in their synthesis. Virology 169:365-376.
Olenina, L. V., T. I. Kuzmina, B. N. Sobolev, T. E. Kuraeva, E. F. Kolesanova, and A. I. Archakov. 2005. Identification of glycosaminoglycan-binding sites within hepatitis C virus envelope glycoprotein E2*. Journal of viral hepatitis 12:584-593.
Owsianka, A., R. F. Clayton, L. D. Loomis-Price, J. A. McKeating, and A. H. Patel. 2001. Functional analysis of hepatitis C virus E2 glycoproteins and virus-like particles reveals structural dissimilarities between different forms of E2. The Journal of general virology 82:1877-1883.
Owsianka, A. M., J. M. Timms, A. W. Tarr, R. J. Brown, T. P. Hickling, A. Szwejk, K. Bienkowska-Szewczyk, B. J. Thomson, A. H. Patel, and J. K. Ball. 2006. Identification of conserved residues in the E2 envelope glycoprotein of the hepatitis C virus that are critical for CD81 binding. Journal of virology 80:8695-8704.
Park, C. Y., H. J. Jun, T. Wakita, J. H. Cheong, and S. B. Hwang. 2009. Hepatitis C virus nonstructural 4B protein modulates sterol regulatory element-binding protein signaling via the AKT pathway. The Journal of biological chemistry 284:9237-9246.
Patterson, B. K., A. Landay, J. N. Siegel, Z. Flener, D. Pessis, A. Chaviano, and R. C. Bailey. 2002. Susceptibility to human immunodeficiency virus-1 infection of human foreskin and cervical tissue grown in explant culture. The American journal of pathology 161:867-873.
Pereira, A. A., and I. M. Jacobson. 2009. New and experimental therapies for HCV. Nature reviews. Gastroenterology & hepatology 6:403-411.
Pileri, P., Y. Uematsu, S. Campagnoli, G. Galli, F. Falugi, R. Petracca, A. J. Weiner, M. Houghton, D. Rosa, G. Grandi, and S. Abrignani. 1998. Binding of hepatitis C virus to CD81. Science 282:938-941.
Ploss, A., M. J. Evans, V. A. Gaysinskaya, M. Panis, H. You, Y. P. de Jong, and C. M. Rice. 2009. Human occludin is a hepatitis C virus entry factor required for infection of mouse cells. Nature 457:882-886.
Pohlmann, S., J. Zhang, F. Baribaud, Z. Chen, G. J. Leslie, G. Lin, A. Granelli-Piperno, R. W. Doms, C. M. Rice, and J. A. McKeating. 2003. Hepatitis C virus glycoproteins interact with DC-SIGN and DC-SIGNR. Journal of virology 77:4070-4080.
Popescu, C. I., and J. Dubuisson. 2009. Role of lipid metabolism in hepatitis C virus assembly and entry. Biology of the cell / under the auspices of the European Cell Biology Organization 102:63-74.
Potel, J., P. Rassam, C. Montpellier, L. Kaestner, E. Werkmeister, B. A. Tews, C. Couturier, C. I. Popescu, T. F. Baumert, E. Rubinstein, J. Dubuisson, P. E. Milhiet, and L. Cocquerel. 2013. EWI-2wint promotes CD81 clustering that abrogates Hepatitis C Virus entry. Cellular microbiology 15:1234-1252.
Powlesland, A. S., T. Fisch, M. E. Taylor, D. F. Smith, B. Tissot, A. Dell, S. Pohlmann, and K. Drickamer. 2008. A novel mechanism for LSECtin binding to Ebola virus surface glycoprotein through truncated glycans. The Journal of biological chemistry 283:593-602.
Ralston, R., K. Thudium, K. Berger, C. Kuo, B. Gervase, J. Hall, M. Selby, G. Kuo, M. Houghton, and Q. L. Choo. 1993. Characterization of hepatitis C virus envelope glycoprotein complexes expressed by recombinant vaccinia viruses. Journal of virology 67:6753-6761.
Rehermann, B. 2009. Hepatitis C virus versus innate and adaptive immune responses: a tale of coevolution and coexistence. The Journal of clinical investigation 119:1745-1754.
Robertson, B., G. Myers, C. Howard, T. Brettin, J. Bukh, B. Gaschen, T. Gojobori, G. Maertens, M. Mizokami, O. Nainan, S. Netesov, K. Nishioka, T. Shin i, P. Simmonds, D. Smith, L. Stuyver, and A. Weiner. 1998. Classification, nomenclature, and database development for hepatitis C virus (HCV) and related viruses: proposals for standardization. International Committee on Virus Taxonomy. Archives of virology 143:2493-2503.
Sato, K., H. Okamoto, S. Aihara, Y. Hoshi, T. Tanaka, and S. Mishiro. 1993. Demonstration of sugar moiety on the surface of hepatitis C virions recovered from the circulation of infected humans. Virology 196:354-357.
Saunier, B., M. Triyatni, L. Ulianich, P. Maruvada, P. Yen, and L. D. Kohn. 2003. Role of the asialoglycoprotein receptor in binding and entry of hepatitis C virus structural proteins in cultured human hepatocytes. Journal of virology 77:546-559.
Scarselli, E., H. Ansuini, R. Cerino, R. M. Roccasecca, S. Acali, G. Filocamo, C. Traboni, A. Nicosia, R. Cortese, and A. Vitelli. 2002. The human scavenger receptor class B type I is a novel candidate receptor for the hepatitis C virus. The EMBO journal 21:5017-5025.
Schwarz, F., and M. Aebi. 2011. Mechanisms and principles of N-linked protein glycosylation. Current opinion in structural biology 21:576-582.
Selby, M. J., E. Glazer, F. Masiarz, and M. Houghton. 1994. Complex processing and protein:protein interactions in the E2:NS2 region of HCV. Virology 204:114-122.
Sen, G. C. 2001. Viruses and interferons. Annual review of microbiology 55:255-281.
Shepard, C. W., L. Finelli, and M. J. Alter. 2005. Global epidemiology of hepatitis C virus infection. The Lancet infectious diseases 5:558-567.
Sherman, M. 2010. Epidemiology of hepatocellular carcinoma. Oncology 78 Suppl 1:7-10.
Shi, S. T., S. J. Polyak, H. Tu, D. R. Taylor, D. R. Gretch, and M. M. Lai. 2002. Hepatitis C virus NS5A colocalizes with the core protein on lipid droplets and interacts with apolipoproteins. Virology 292:198-210.
Shimizu, Y. K., S. M. Feinstone, M. Kohara, R. H. Purcell, and H. Yoshikura. 1996. Hepatitis C virus: detection of intracellular virus particles by electron microscopy. Hepatology 23:205-209.
Spaete, R. R., D. Alexander, M. E. Rugroden, Q. L. Choo, K. Berger, K. Crawford, C. Kuo, S. Leng, C. Lee, R. Ralston, and et al. 1992. Characterization of the hepatitis C virus E2/NS1 gene product expressed in mammalian cells. Virology 188:819-830.
Stockert, R. J. 1995. The asialoglycoprotein receptor: relationships between structure, function, and expression. Physiological reviews 75:591-609.
Takebe, Y., C. J. Saucedo, G. Lund, R. Uenishi, S. Hase, T. Tsuchiura, N. Kneteman, K. Ramessar, D. L. Tyrrell, M. Shirakura, T. Wakita, J. B. McMahon, and B. R. O''Keefe. 2013. Antiviral lectins from red and blue-green algae show potent in vitro and in vivo activity against hepatitis C virus. PloS one 8:e64449.
Tellinghuisen, T. L., M. J. Evans, T. von Hahn, S. You, and C. M. Rice. 2007. Studying hepatitis C virus: making the best of a bad virus. Journal of virology 81:8853-8867.
Tellinghuisen, T. L., and C. M. Rice. 2002. Interaction between hepatitis C virus proteins and host cell factors. Current opinion in microbiology 5:419-427.
Thomson, B. J. 2009. Hepatitis C virus: the growing challenge. British medical bulletin 89:153-167.
Thomssen, R., S. Bonk, and A. Thiele. 1993. Density heterogeneities of hepatitis C virus in human sera due to the binding of beta-lipoproteins and immunoglobulins. Medical microbiology and immunology 182:329-334.
Tilg, H. 1997. New insights into the mechanisms of interferon alfa: an immunoregulatory and anti-inflammatory cytokine. Gastroenterology 112:1017-1021.
Timpe, J. M., Z. Stamataki, A. Jennings, K. Hu, M. J. Farquhar, H. J. Harris, A. Schwarz, I. Desombere, G. L. Roels, P. Balfe, and J. A. McKeating. 2008. Hepatitis C virus cell-cell transmission in hepatoma cells in the presence of neutralizing antibodies. Hepatology 47:17-24.
van de Laar, T. J., G. V. Matthews, M. Prins, and M. Danta. 2010. Acute hepatitis C in HIV-infected men who have sex with men: an emerging sexually transmitted infection. AIDS 24:1799-1812.
Varki A, Cummings RD, Esko JD, Freeze HH, Stanley P, Bertozzi CR, Hart GW, and e. Etzler ME. 2009. Essentials of Glycobiology. 2nd edn. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press
Vieyres, G., X. Thomas, V. Descamps, G. Duverlie, A. H. Patel, and J. Dubuisson. 2010. Characterization of the envelope glycoproteins associated with infectious hepatitis C virus. Journal of virology 84:10159-10168.
Wakita, T., T. Pietschmann, T. Kato, T. Date, M. Miyamoto, Z. Zhao, K. Murthy, A. Habermann, H. G. Krausslich, M. Mizokami, R. Bartenschlager, and T. J. Liang. 2005. Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nature medicine 11:791-796.
Wertheimer, A. M., C. Miner, D. M. Lewinsohn, A. W. Sasaki, E. Kaufman, and H. R. Rosen. 2003. Novel CD4+ and CD8+ T-cell determinants within the NS3 protein in subjects with spontaneously resolved HCV infection. Hepatology 37:577-589.
Wiegand, J., P. Buggisch, W. Boecher, S. Zeuzem, C. M. Gelbmann, T. Berg, W. Kauffmann, B. Kallinowski, M. Cornberg, E. Jaeckel, H. Wedemeyer, and M. P. Manns. 2006. Early monotherapy with pegylated interferon alpha-2b for acute hepatitis C infection: the HEP-NET acute-HCV-II study. Hepatology 43:250-256.
Wojtowicz, K., W. Szaflarski, R. Januchowski, P. Zawierucha, M. Nowicki, and M. Zabel. 2012. Inhibitors of N-glycosylation as a potential tool for analysis of the mechanism of action and cellular localisation of glycoprotein P. Acta biochimica Polonica 59:445-450.
Wolk, B., D. Sansonno, H. G. Krausslich, F. Dammacco, C. M. Rice, H. E. Blum, and D. Moradpour. 2000. Subcellular localization, stability, and trans-cleavage competence of the hepatitis C virus NS3-NS4A complex expressed in tetracycline-regulated cell lines. Journal of virology 74:2293-2304.
Yabe, R., H. Tateno, and J. Hirabayashi. 2010. Frontal affinity chromatography analysis of constructs of DC-SIGN, DC-SIGNR and LSECtin extend evidence for affinity to agalactosylated N-glycans. The FEBS journal 277:4010-4026.
Yang, J. D., and L. R. Roberts. 2010. Hepatocellular carcinoma: A global view. Nature reviews. Gastroenterology & hepatology 7:448-458.
Young, K. C., K. L. Lindsay, K. J. Lee, W. C. Liu, J. W. He, S. L. Milstein, and M. M. Lai. 2003. Identification of a ribavirin-resistant NS5B mutation of hepatitis C virus during ribavirin monotherapy. Hepatology 38:869-878.
Yu, C. I., and B. L. Chiang. 2010. A new insight into hepatitis C vaccine development. Journal of biomedicine & biotechnology 2010:548280.