Parvovirus B19 is the causative agent for fifth disease. Recently, it was reported that B19 may play an important role in the pathogenesis of HBV in Vietnamese patients. We here aimed to investigate whether B19 infection may be a co-factor for disease progression in European patients with chronic hepatitis C.
In this study, 91 serum samples from well characterized and histologically staged chronic hepatitis C patients and 50 serum samples from chronic hepatitis B patients were investigated for the presence of B19 antibodies and B19 DNA (real-time PCR). Moreover, 50 explanted livers and 32 liver biopsy samples (obtained for routine staging of chronic liver disease) were tested for B19 DNA. Finally, B19-specific CD4+ T cell responses were studied in 13 anti-B19-positive hepatitis C patients and 19 serologically recovered B19 infected healthy individuals.
In contrast to the previous study on Vietnamese HBV-patients, we detected B19 DNA in only 1/ 91 serum samples from chronic hepatitis C patients and in none of the serum samples from chronic hepatitis B patients. B19 IgG antibodies were found in 67/91 (74%) HCV patients, while only the B19 DNA-positive hepatitis C patient was anti-B19-IgM positive. Clinical and histological characteristics did not differ between anti-B19-IgG-positive and IgG-negative patients. Surprisingly, B19 DNA was amplified from more than half of the liver tissues studied and more frequently detectable in explanted end-stage liver tissues (74%) than in biopsy samples (44%)(p<0.05). However, there was no significant difference in virus copy number per cell between these two groups. Finally, B19-specific CD4+ T cell responses were detected in a similar frequency in healthy anti-B19-positive individuals (3/19; 16%) and chronic hepatitis C patients (3/13; 23%).
In conclusion, this study gives evidence that B19 may persist in the liver.
However, even though B19-DNA can be detected intrahepatically, there is no evidence that B19 is a “hepatitis virus” worsening liver disease and accelerating disease progression of chronic hepatitis C in European patients.
Role of Parvovirus B19 infection in hepatitis C -- Reference
6. Reference
1. Choo, Q.L., et al., Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science, 1989. 244(4902): p. 359-62.
2. Touzet, S., et al., Epidemiology of hepatitis C virus infection in seven European Union countries: a critical analysis of the literature. HENCORE Group. (Hepatitis C European Network for Co-operative Research. Eur J Gastroenterol Hepatol, 2000. 12(6): p. 667-78.
3. Manns, M.P., H. Wedemeyer, and M. Cornberg, Treating viral hepatitis C:
efficacy, side effects, and complications. Gut, 2006. 55(9): p. 1350-9.
4. Rehermann, B. and M. Nascimbeni, Immunology of hepatitis B virus and hepatitis C virus infection. Nat Rev Immunol, 2005. 5(3): p. 215-29.
5. Evans, M.J., et al., Claudin-1 is a hepatitis C virus co-receptor required for a late step in entry. Nature, 2007. 446(7137): p. 801-5.
6. Matos, C.A., et al., Steatosis in chronic hepatitis C: relationship to the virus and host risk factors. J Gastroenterol Hepatol, 2006. 21(8): p. 1236-9.
7. Lonardo, A., et al., Hepatitis C and steatosis: a reappraisal. J Viral Hepat, 2006. 13(2): p. 73-80.
8. Rumi, M.G., et al., Hepatitis C reactivation in patients with chronic infection with genotypes 1b and 2c: a retrospective cohort study of 206 untreated patients. Gut, 2005. 54(3): p. 402-6.
9. Bruno, S., et al., Hepatitis C virus genotype 1b as a major risk factor associated with hepatocellular carcinoma in patients with cirrhosis: a
seventeen-year prospective cohort study. Hepatology, 2007. 46(5): p. 1350-6.
10. Thimme, R., et al., Determinants of viral clearance and persistence during acute hepatitis C virus infection. J Exp Med, 2001. 194(10): p. 1395-406.
11. Wedemeyer, H., et al., Impaired effector function of hepatitis C virus-specific CD8+ T cells in chronic hepatitis C virus infection. J Immunol, 2002. 169(6):
p. 3447-58.
12. Diepolder, H.M., et al., Possible mechanism involving T-lymphocyte response to non-structural protein 3 in viral clearance in acute hepatitis C virus infection. Lancet, 1995. 346(8981): p. 1006-7.
13. Lechner, F., et al., Analysis of successful immune responses in persons infected with hepatitis C virus. J Exp Med, 2000. 191(9): p. 1499-512.
14. Kim, A.Y., et al., Impaired hepatitis C virus-specific T cell responses and recurrent hepatitis C virus in HIV coinfection. PLoS Med, 2006. 3(12): p.
e492.
15. Strickland, G.T., Liver disease in Egypt: hepatitis C superseded
schistosomiasis as a result of iatrogenic and biological factors. Hepatology, 2006. 43(5): p. 915-22.
16. Welsh, R.M., et al., The privacy of T cell memory to viruses. Curr Top Microbiol Immunol, 2006. 311: p. 117-53.
17. Selin, L.K., et al., CD8 memory T cells: cross-reactivity and heterologous immunity. Semin Immunol, 2004. 16(5): p. 335-47.
18. Jaeckel, E., et al., Treatment of acute hepatitis C with interferon alfa-2b. N Engl J Med, 2001. 345(20): p. 1452-7.
19. Wiegand, J., et al., Early monotherapy with pegylated interferon alpha-2b for acute hepatitis C infection: the HEP-NET acute-HCV-II study. Hepatology, 2006. 43(2): p. 250-6.
20. Cossart, Y.E., et al., Parvovirus-like particles in human sera. Lancet, 1975.
1(7898): p. 72-3.
Role of Parvovirus B19 infection in hepatitis C -- Reference
21. Anderson, L.J., et al., Detection of antibodies and antigens of human parvovirus B19 by enzyme-linked immunosorbent assay. J Clin Microbiol, 1986. 24(4): p. 522-6.
22. Cohen, B.J. and M.M. Buckley, The prevalence of antibody to human
parvovirus B19 in England and Wales. J Med Microbiol, 1988. 25(2): p. 151-3.
23. Kelly, H.A., et al., The age-specific prevalence of human parvovirus immunity in Victoria, Australia compared with other parts of the world. Epidemiol Infect, 2000. 124(3): p. 449-57.
24. Tsujimura, M., et al., Human parvovirus B19 infection in blood donors. Vox Sang, 1995. 69(3): p. 206-12.
25. Broliden, K., T. Tolfvenstam, and O. Norbeck, Clinical aspects of parvovirus B19 infection. J Intern Med, 2006. 260(4): p. 285-304.
26. Lindblom, A., et al., Slow clearance of human parvovirus B19 viremia following acute infection. Clin Infect Dis, 2005. 41(8): p. 1201-3.
27. Ozawa, K., et al., The gene encoding the nonstructural protein of B19 (human) parvovirus may be lethal in transfected cells. J Virol, 1988. 62(8): p. 2884-9.
28. Moffatt, S., et al., Human parvovirus B19 nonstructural (NS1) protein induces apoptosis in erythroid lineage cells. J Virol, 1998. 72(4): p. 3018-28.
29. Ozawa, K. and N. Young, Characterization of capsid and noncapsid proteins of B19 parvovirus propagated in human erythroid bone marrow cell cultures.
J Virol, 1987. 61(8): p. 2627-30.
30. Shade, R.O., et al., Nucleotide sequence and genome organization of human parvovirus B19 isolated from the serum of a child during aplastic crisis. J Virol, 1986. 58(3): p. 921-36.
31. Zadori, Z., et al., A viral phospholipase A2 is required for parvovirus infectivity. Dev Cell, 2001. 1(2): p. 291-302.
32. Brown, K.E., S.M. Anderson, and N.S. Young, Erythrocyte P antigen: cellular receptor for B19 parvovirus. Science, 1993. 262(5130): p. 114-7.
33. Weigel-Kelley, K.A., M.C. Yoder, and A. Srivastava, Recombinant human parvovirus B19 vectors: erythrocyte P antigen is necessary but not sufficient for successful transduction of human hematopoietic cells. J Virol, 2001. 75(9):
p. 4110-6.
34. Weigel-Kelley, K.A., M.C. Yoder, and A. Srivastava, Alpha5beta1 integrin as a cellular coreceptor for human parvovirus B19: requirement of functional activation of beta1 integrin for viral entry. Blood, 2003. 102(12): p. 3927-33.
35. Munakata, Y., et al., Ku80 autoantigen as a cellular coreceptor for human parvovirus B19 infection. Blood, 2005. 106(10): p. 3449-56.
36. Servant, A., et al., Genetic diversity within human erythroviruses:
identification of three genotypes. J Virol, 2002. 76(18): p. 9124-34.
37. Umene, K. and T. Nunoue, The genome type of human parvovirus B19 strains isolated in Japan during 1981 differs from types detected in 1986 to 1987: a correlation between genome type and prevalence. J Gen Virol, 1990. 71 (Pt 4):
p. 983-6.
38. Umene, K. and T. Nunoue, Genetic diversity of human parvovirus B19 determined using a set of restriction endonucleases recognizing four or five base pairs and partial nucleotide sequencing: use of sequence variability in virus classification. J Gen Virol, 1991. 72 (Pt 8): p. 1997-2001.
39. Kurtzman, G.J., et al., Immune response to B19 parvovirus and an antibody defect in persistent viral infection. J Clin Invest, 1989. 84(4): p. 1114-23.
Role of Parvovirus B19 infection in hepatitis C -- Reference
40. von Poblotzki, A., et al., Lymphoproliferative responses after infection with human parvovirus B19. J Virol, 1996. 70(10): p. 7327-30.
41. Tolfvenstam, T., et al., Mapping of B-cell epitopes on human parvovirus B19 non-structural and structural proteins. Vaccine, 2000. 19(7-8): p. 758-63.
42. Isa, A., et al., Prolonged activation of virus-specific CD8+T cells after acute B19 infection. PLoS Med, 2005. 2(12): p. e343.
43. Norbeck, O., et al., Sustained CD8+ T-cell responses induced after acute parvovirus B19 infection in humans. J Virol, 2005. 79(18): p. 12117-21.
44. Franssila, R., et al., T helper cell-mediated interferon-gamma expression after human parvovirus B19 infection: persisting VP2-specific and transient VP1u-specific activity. Clin Exp Immunol, 2005. 142(1): p. 53-61.
45. Franssila, R. and K. Hedman, T-helper cell-mediated interferon-gamma, interleukin-10 and proliferation responses to a candidate recombinant vaccine for human parvovirus B19. Vaccine, 2004. 22(27-28): p. 3809-15.
46. Lindner, J., et al., CD4(+) T-cell responses against the VP1-unique region in individuals with recent and persistent parvovirus B19 infection. J Vet Med B Infect Dis Vet Public Health, 2005. 52(7-8): p. 356-61.
47. Kasprowicz, V., et al., Tracking of peptide-specific CD4+ T-cell responses after an acute resolving viral infection: a study of parvovirus B19. J Virol, 2006. 80(22): p. 11209-17.
48. Rehermann, B., et al., The hepatitis B virus persists for decades after patients' recovery from acute viral hepatitis despite active maintenance of a cytotoxic T-lymphocyte response. Nat Med, 1996. 2(10): p. 1104-8.
49. Isa, A., et al., Aberrant cellular immune responses in humans infected persistently with parvovirus B19. J Med Virol, 2006. 78(1): p. 129-33.
50. Harris, J.W., Parvovirus B19 for the hematologist. Am J Hematol, 1992. 39(2):
p. 119-30.
51. Young, N.S. and K.E. Brown, Parvovirus B19. N Engl J Med, 2004. 350(6): p.
586-97.
52. Heegaard, E.D. and K.E. Brown, Human parvovirus B19. Clin Microbiol Rev, 2002. 15(3): p. 485-505.
53. Kuhl, U., et al., Interferon-beta treatment eliminates cardiotropic viruses and improves left ventricular function in patients with myocardial persistence of viral genomes and left ventricular dysfunction. Circulation, 2003. 107(22): p.
2793-8.
54. Eis-Hubinger, A.M., et al., Evidence for persistence of parvovirus B19 DNA in livers of adults. J Med Virol, 2001. 65(2): p. 395-401.
55. Abe, K., et al., Characterization of erythrovirus B19 genomes isolated in liver tissues from patients with fulminant hepatitis and biliary atresia who
underwent liver transplantation. Int J Med Sci, 2007. 4(2): p. 105-9.
56. He, Z., et al., Retrospective analysis of non-A-E hepatitis: possible role of hepatitis B and C virus infection. J Med Virol, 2003. 69(1): p. 59-65.
57. Wong, S., N.S. Young, and K.E. Brown, Prevalence of parvovirus B19 in liver tissue: no association with fulminant hepatitis or hepatitis-associated aplastic anemia. J Infect Dis, 2003. 187(10): p. 1581-6.
58. Drago, F., et al., Parvovirus B19 infection associated with acute hepatitis and a purpuric exanthem. Br J Dermatol, 1999. 141(1): p. 160-1.
59. Hillingso, J.G., I.P. Jensen, and L. Tom-Petersen, Parvovirus B19 and acute hepatitis in adults. Lancet, 1998. 351(9107): p. 955-6.
Role of Parvovirus B19 infection in hepatitis C -- Reference
60. Karetnyi, Y.V., et al., Human parvovirus B19 infection in acute fulminant liver failure. Arch Virol, 1999. 144(9): p. 1713-24.
61. Sokal, E.M., et al., Acute parvovirus B19 infection associated with fulminant hepatitis of favourable prognosis in young children. Lancet, 1998. 352(9142):
p. 1739-41.
62. Yoto, Y., et al., Human parvovirus B19 infection associated with acute hepatitis. Lancet, 1996. 347(9005): p. 868-9.
63. So, K., et al., Urgent liver transplantation for acute liver failure due to parvovirus B19 infection complicated by primary Epstein-Barr virus and cytomegalovirus infections and aplastic anaemia. Intern Med J, 2007. 37(3): p.
192-5.
64. Bultmann, B.D., et al., Fatal parvovirus B19-associated myocarditis clinically mimicking ischemic heart disease: an endothelial cell-mediated disease. Hum Pathol, 2003. 34(1): p. 92-5.
65. Wandinger, K., et al., Association between clinical disease activity and Epstein-Barr virus reactivation in MS. Neurology, 2000. 55(2): p. 178-84.
66. Gan, Y.J., et al., A defective, rearranged Epstein-Barr virus genome in EBER-negative and EBER-positive Hodgkin's disease. Am J Pathol, 2002. 160(3): p.
781-6.
67. Crawford, M.P., et al., High prevalence of autoreactive, neuroantigen-specific CD8+ T cells in multiple sclerosis revealed by novel flow cytometric assay.
Blood, 2004. 103(11): p. 4222-31.
68. Semmo, N., et al., Analysis of the relationship between cytokine secretion and proliferative capacity in hepatitis C virus infection. J Viral Hepat, 2007. 14(7):
p. 492-502.
69. Toan, N.L., et al., Co-infection of human parvovirus B19 in Vietnamese patients with hepatitis B virus infection. J Hepatol, 2006. 45(3): p. 361-9.
70. Hsu, T.C., et al., Human parvovirus B19 infection in patients with chronic hepatitis B or hepatitis C infection. J Gastroenterol Hepatol, 2005. 20(5): p.
733-8.
71. Anderson, M.J., et al., Experimental parvoviral infection in humans. J Infect Dis, 1985. 152(2): p. 257-65.
72. Anderson, M.J., et al., Human parvovirus, the cause of erythema infectiosum (fifth disease)? Lancet, 1983. 1(8338): p. 1378.
73. Naides, S.J., Infection with Parvovirus B19. Curr Infect Dis Rep, 1999. 1(3): p.
273-278.
74. Katz, V.L., N.C. Chescheir, and M. Bethea, Hydrops fetalis from B19 parvovirus infection. J Perinatol, 1990. 10(4): p. 366-8.
75. Gilbert, G.L., Parvovirus B19 infection and its significance in pregnancy.
Commun Dis Intell, 2000. 24 Suppl: p. 69-71.
76. Orth, T., et al., Human parvovirus B19 infection associated with severe acute perimyocarditis in a 34-year-old man. Eur Heart J, 1997. 18(3): p. 524-5.
77. Papadogiannakis, N., et al., Active, fulminant, lethal myocarditis associated with parvovirus B19 infection in an infant. Clin Infect Dis, 2002. 35(9): p.
1027-31.
78. Brown, K.E., Haematological consequences of parvovirus B19 infection.
Baillieres Best Pract Res Clin Haematol, 2000. 13(2): p. 245-59.
79. Koch, W.C., et al., Manifestations and treatment of human parvovirus B19 infection in immunocompromised patients. J Pediatr, 1990. 116(3): p. 355-9.
Role of Parvovirus B19 infection in hepatitis C -- Reference
80. Schmidt, M., et al., Blood donor screening for parvovirus B19 in Germany and Austria. Transfusion, 2007. 47(10): p. 1775-82.
81. Cassinotti, P., et al., Evidence for persistence of human parvovirus B19 DNA in bone marrow. J Med Virol, 1997. 53(3): p. 229-32.
82. Heegaard, E.D., et al., Prevalence of parvovirus B19 and parvovirus V9 DNA and antibodies in paired bone marrow and serum samples from healthy individuals. J Clin Microbiol, 2002. 40(3): p. 933-6.
83. Lundqvist, A., et al., Clinical and laboratory findings in immunocompetent patients with persistent parvovirus B19 DNA in bone marrow. Scand J Infect Dis, 1999. 31(1): p. 11-6.
84. Lundqvist, A., et al., Prevalence of parvovirus B19 DNA in bone marrow of patients with haematological disorders. Scand J Infect Dis, 1999. 31(2): p.
119-22.
85. Chevrel, G., et al., Dermatomyositis associated with the presence of parvovirus B19 DNA in muscle. Rheumatology (Oxford), 2000. 39(9): p.
1037-9.
86. Nikkari, S., et al., Persistence of parvovirus B19 in synovial fluid and bone marrow. Ann Rheum Dis, 1995. 54(7): p. 597-600.
87. Vuorinen, T., et al., Presence of parvovirus B19 DNA in chronic urticaric and healthy human skin. J Clin Virol, 2002. 25(2): p. 217-21.
88. Kuhl, U., Antiviral treatment of myocarditis and acute dilated cardiomyopathy.
Heart Fail Clin, 2005. 1(3): p. 467-74.
89. Langnas, A.N., et al., Parvovirus B19 as a possible causative agent of fulminant liver failure and associated aplastic anemia. Hepatology, 1995.
22(6): p. 1661-5.
90. Pinho, J.R., et al., Detection of human parvovirus B19 in a patient with hepatitis. Braz J Med Biol Res, 2001. 34(9): p. 1131-8.
91. Franssila, R., K. Hokynar, and K. Hedman, T helper cell-mediated in vitro responses of recently and remotely infected subjects to a candidate
recombinant vaccine for human parvovirus b19. J Infect Dis, 2001. 183(5): p.
805-9.
92. Mitchell, L.A., R. Leong, and K.A. Rosenke, Lymphocyte recognition of human parvovirus B19 non-structural (NS1) protein: associations with occurrence of acute and chronic arthropathy? J Med Microbiol, 2001. 50(7):
p. 627-35.
93. Bain, C., et al., Impaired allostimulatory function of dendritic cells in chronic hepatitis C infection. Gastroenterology, 2001. 120(2): p. 512-24.
94. Aberle, J.H., et al., CD4+ T cell responses in patients with chronic hepatitis C undergoing peginterferon/ribavirin therapy correlate with faster, but not sustained, viral clearance. J Infect Dis, 2007. 195(9): p. 1315-9.
95. Meyer, M.F., et al., Clearance of low levels of HCV viremia in the absence of a strong adaptive immune response. Virol J, 2007. 4: p. 58.
96. Wedemeyer, H., et al., Cross-reactivity between hepatitis C virus and
Influenza A virus determinant-specific cytotoxic T cells. J Virol, 2001. 75(23):
p. 11392-400.
Role of Parvovirus B19 infection in hepatitis C -- Abbreviations
7. List of abbreviations:
B19 Parvovirus B19
HCV Hepatitis C virus
qPCR Quantity Polymerase chain reaction
nPCR Nested Polymerase chain reaction
FH Fulminant hepatitis
PBMC Peripheral blood mononuclear cell
ALT alanine aminotransferase
AST aspartate aminotransferase
gGT gamma-glutamyltransferase
ULM Upper limited of normal
AHB Acute hepatitis B
CHB Chronic hepatitis B
LC Liver cirrhosis
HCC Hepatocellular carcinoma
VAHS Virus-associated-hemophagocytic
syndrome
NASH Nonalcoholic steatohepatitis
DMSO dimethyl sulfoxide
PHA phytohemagglutinin
TT Tetanus toxoid
Role of Parvovirus B19 infection in hepatitis C -- Acknowledgements
8. Appendix
Acknowledgements
I would like to acknowledge numerous people for their kindness and support in the past two years. Without their priceless help, this thesis could not be finished.
At the very beginning, I’m honored to express my appreciation to Prof. Dr. med.
Michael P. Manns, who gave me great support during my two-year advanced study in his Department.
Here I must express the deepest gratitude to my supervisor, Priv. Doz. Dr. med.
Heiner Wedemeyer. He gave me the opportunity to join his lab. He offered me valuable ideas, suggestions and guidance with his professional attainment in Hepatology and Immunology. His patience and kindness are greatly appreciated.
Moreover, he always puts high priority on my project and has provided me with good communication whenever he is available. I’m very much obliged to his efforts of helping me complete the dissertation.
I’m also grateful to PD. Dr. med. Albert Heim and PD. Dr. rer. nat. C.-Thomas Bock who gave me patient guidance and many useful advices and supports.
I owe special thanks to all members of our group: Markus, Katja, Sandra, Evi, Verana, Kerstin, Suneetha. All of them gave me great supports and showed warmest concern not only for my study but also for my daily life in Germany.
I wish to extend my gratitude to Hong Jiang, Regina, Peter, Birgit, Ursel, Nicole and many many faculty members and staffs of the department assisted and encouraged me in various ways during my study.
At last, I would like to say thanks to my family and my husband for their support and love.
Role of Parvovirus B19 infection in hepatitis C -- CV
Curriculum vitae
Personal information:
Name: Wang
First name: Chun
Gender: Female
Birthday: 06.06.1970
Birthplace: Shanghai, P.R. China
Nation: Chinese
Education:
Sept. 1977- July. 1982 Elementary education Sept. 1982- July.1988 High school
Study:
Sept.1988- July.1993 Shanghai Tiedao Medical College, Shanghai, China Completion: medical bachelor’s degree
Sept.2001- July. 2004 Tongji University, Shanghai, China Completion: medical master’s degree Occupation:
Aug. 1993- Aug. 2001 Shanghai Railway Bureau Central Hospital Dept. Internal medicine and Dept. Endocrinology Shanghai, China
Aug. 2004-Sept. 2005 Tongji University affiliated Shanghai Tenth People´s Hospital.
Dept. Endocrinology Shanghai, China
Oct. 2005-Sept. 2007 Medical school Hannover,
Dept. Gastroenterology, Hepatology and Endocrinology
Hannover, Germany
Role of Parvovirus B19 infection in hepatitis C -- Erklärung
Erklärung
Laut Paragraf 2, Absatz 2, Nummern 5 und 6 der Promotionsordnung der Medizinischen Hochschule Hannover.
Ich erkläre, dass ich die der Medizinischen Hochschule Hannover zur Promotion eingereichte Dissertation mit dem Titel „Parvo-Virus B19 Infection: Evidence for intrahepatic long-term persistence but no association with disease progression in chronic hepatitis C" in der Klinik für Gastroenterologie, Hepatologie und Endokrinologie
der Medizinischen Hochschule Hannover unter Leitung von Priv. Doz. Dr. med.
Heiner Wedemeyer ohne sonstige Hilfe selbst durchgeführt und bei der Abfassung der Dissertation keine anderen als die dort aufgeführten Hilfsmittel benutzt habe.
Ich habe diese Dissertation bisher an keiner in- oder ausländischen Hochschule zur Promotion eingereicht. Weiterhin versichere ich, dass ich den beantragten Titel bisher noch nicht erworben habe.
Hannover, den 3.2.2008
Chun Wang