Skip to main content
SpringerLink
Log in

Identification of β-Subunit of Bacterial RNA-Polymerase—A Non-Species-Specific Bacterial Protein—As Target of Antibodies in Primary Biliary Cirrhosis

  • Published:
Digestive Diseases and Sciences Aims and scope Submit manuscript

Abstract

Several observations suggest that bacteria induce autoimmunity in primary biliary cirrhosis (PBC). Since no PBC-specific bacterial species could be identified, it can be speculated that the triggers are non-species-specific bacterial proteins. This hypothesis would imply that several or even all bacterial species can trigger PBC. Therefore, we investigated whether PBC exhibits immune reactions to non-species-specific bacterial antigens. Yersinia enterocolitica O3 was screened for the presence of proteins that were labeled by immunoblotting using PBC sera. We focused our investigations on a 160-kDa protein, which was further enriched and characterized by partial N-terminal amino acid sequencing. The prevalence of antibodies to this protein was determined by immunoblotting in a variety of diseases. The 160-kDa protein was identified as the β-subunit of bacterial RNA-polymerase, a highly conserved bacterial protein with a very high degree of sequence identity among all bacterial species. Antibodies to the β-subunit of bacterial RNA polymerase were specific for this protein. Until now no mammalian protein could be found that cross-reacts with these antibodies. The prevalence of antibodies to the β-subunit of bacterial RNA polymerase (ARPA) using the protein from Yersinia enterocolitica O3 (serum dilution 1:1000) was: healthy controls (HC, N = 101) 7.9%, primary biliary cirrhosis (PBC, N = 61) 32.8%, autoimmune hepatitis type 1 (AIH, N = 46) 26.1%, alcoholic liver cirrhosis (ALC, N = 44) 9.1%, Crohn's disease (CD, N = 38) 7.9%, ulcerative colitis (UC, N = 24) 8.3%, primary sclerosing cholangitis + UC (PSC/UC, N = 11) 0%, acute yersiniosis (Yers, N = 36) 19.4%, acute infection with Campylobacter jejuni (Camp, N = 10) 0%, acute Q-fever (QF, N = 16) 6.25%, chronic hepatitis C (HCV, N = 39) 7.7%, c-ANCA-positive vasculitis (Vasc, N = 40) 15%, systemic lupus erythematosus (SLE, N = 28) 10.7%, and malaria tropica (MT, N = 24) 16.7%. There was no significant difference between PBC and AIH. The group of autoimmune liver diseases (PBC + AIH, N = 107, 29.9%) differed highly significantly from HC, chronic inflammatory bowel diseases (CD + UC + PSC/UC, N = 73, 6.8%), ALC, and HCV and also differed significantly (P = 0.01) from the group with bacterial and parasitic diseases (Yers + Camp + QF + MT, N = 86, 13.95%) and from the group with Vasc + SLE (N = 68, 13.2%). Testing of ARPA using the protein from E. coli yielded nearly identical results. In conclusion, an increased prevalence of antibodies to the β-subunit of bacterial RNA polymerase, a highly conserved non-species-specific bacterial protein, can be found in primary biliary cirrhosis, but also in autoimmune hepatitis type I. These findings do not add an argument for a bacterial trigger of PBC. Rather, they suggest that ARPA belong to the pool of natural antibodies that are up-regulated in autoimmune liver diseases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Burroughs AK, Butler P, Sternberg MJ, Baum H: Molecular mimicry in liver disease. Nature 358:377-378, 1992

    Google Scholar 

  2. Albert LJ, Inman RD: Molecular mimicry and autoimmunity. N Engl J Med 27:2068-2074, 2000

    Google Scholar 

  3. Baum H, Berg PA: The complex nature of mitochondrial antibodies and their relation to primary biliary cirrhosis. Semin Liver Dis, 1:309-321, 1981

    Google Scholar 

  4. Gershwin ME, Mackay IR, Sturgess A, Coppel RL: Identification and specifity of a cDNA encoding the 70 kDa mitochondrial antigen recognized in primary biliary cirrhosis. J Immunol 138:3525-3531, 1987

    Google Scholar 

  5. Berg PA, Klein R: Cholestatic diseases of the liver. Cur Opin in Gastroenterol 12:258-271, 1996

    Google Scholar 

  6. Leung PSC, Coppel RL, Ansari A, Munoz S, Gershwin ME: Animitochondrial antibodies in primary biliary cirrhosis. Semin Liver Dis 17:61-69, 1997

    Google Scholar 

  7. Gershwin ME, Mackay IR, Sturgess A, Coppel RL: Identification and specificity of a c-DNA encoding the 70 kd mitochondrial antigen recognized in primary biliary cirrhosis. J Immunol 138:3525-3531, 1978

    Google Scholar 

  8. Strassburg CP, Jaeckel E, Manns MP: Anti-mitochondrial antibodies and other immunological tests in primary biliary cirrhosis. Eur J Gastroenterol Hepatol 11:595-601, 1999

    Google Scholar 

  9. Joplin RE, Neuberger JM: Immunopathology of primary biliary cirrhosis. Eur J Gastroenterol Hepatol 11:587-594, 1999

    Google Scholar 

  10. Quaranta S, Van de Water J, Ishibashi H, Rosina F, Coppel R, Uibo R, Gershwin ME: The immunopathogenesis of primary biliary cirrhosis. Hepato-Gastroenterology 46:3041-3047, 1999

    Google Scholar 

  11. Van de Water JA, Ansari A, Prindiville T, Coppel RL, Ricalton N, Kotzin BL, Liu S, Roche TE, Krams SM, Munoz S, Gershwin ME: Heterogeneity of autoreactive T-cell clones specific for the E2 component of the pyruvate dehydrogenase complex in primary biliary cirrhosis. J Exp Med 181:723-733, 1995

    Google Scholar 

  12. Loehr H, Fleischer B, Gerken G, Yeaman SJ, Meyer zum Bueschenfelde KH, Manns M: Autoreactive liver-infiltrating T cells in primary biliary cirrhosis recognize inner mitochondrial epitopes and the pyruvate dehydrogenase complex. J Hepatol 18:322-327, 1993

    Google Scholar 

  13. Palmer JM, Diamond AG, Yeaman SJ, Bassendine MF, Jones DEJ: T cell responses to the putative dominant autoepitope in primary biliary cirrhosis (PBC). Clin Exp Immunol 116:133-139, 1999

    Google Scholar 

  14. Van de Water J, Shimoda S, Niho Y, Coppel R, Ansari A, Gershwin ME: The role of T cells in primary biliary cirrhosis. Semin Liver Dis 17:105-113, 1997

    Google Scholar 

  15. Agarwal K, Jones DEJ, Bassendine MF: Genetic susceptibility to primary biliary cirrhosis. Eur J Gastroenterol Hepatol 11:603-606, 1999

    Google Scholar 

  16. Haydon GH, Neuberger J: PBC: an infectious disease? Gut 47:586-588, 2000

    Google Scholar 

  17. Burroughs AK, Rosenstein IJ, Epstein O, Hamilton-Miller JM, Brumfitt W, Sherlock S: Bacteriuria and primary biliary cirrhosis. Gut 25:133-137, 1984

    Google Scholar 

  18. Butler P, Hamilton Miller J, Baum H, Burroughs AK: Detection of M2 antibodies in patients with recurrent urinary tract infection using an ELISA and purified PBC specific antigens. Evidence for a molecular mimicry mechanism in the pathogenesis of primary biliary cirrhosis? Biochem Mol Biol Int 35:473-485, 1995

    Google Scholar 

  19. Rosenstein IJ, Hazlehurst GR, Burroughs AK, Epstein O, Sherlock S, Brumfitt W: Recurrent bacteriuria and primary biliary cirrhosis: ABO blood group, P1 blood group and secretor status. J Clin Pathol 37:1055-1058, 1984

    Google Scholar 

  20. Stemerowicz R, Hopf U, Moller B, Wittenbrink C, Rodloff A, Reinhard R, Freudenberg M, Galano SC: Are mitochondrial antibodies in primary biliary cirrhosis induced by R-(rough)-mutants of enterobacteriaceae? Lancet 2:1166-1170, 1988

    Google Scholar 

  21. Klein R, Wiebel M, Engelhard S, Berg PA: Sera from patients with tuberculosis recognize the M2a-epitope (E2-subunit of pyruvate dehydrogenase) specific for primary biliary cirrhosis. Clin Exp Immunol 92:308-316, 1993

    Google Scholar 

  22. Vilagut L, Vila J, Vinas O; Pares A, Gines A, Jimenez de Anta MT, Rodes J: Cross-reactivity of anti-Mycobacterium gordonae antibodies with the major mitochondrial autoantigen in primary biliary cirrhosis. J Hepatol 21:673-677, 1994

    Google Scholar 

  23. Vilagut L, Pares A, Vinas O, Vila J, Jimenez de Anta T, Rodes J: Antibodies to mycobacterial 65-kD heat shock protein cross-react with the main mitochondrial antigens in patients with primary biliary cirrhosis. Eur J Clin Invest 27:667-672, 1997

    Google Scholar 

  24. O'Donohue J, McFarlane B, Bomford A, Yates M, Williams R: Antibodies to atypical mycobacteria in primary biliary cirrhosis. J Hepatol 21:887-889, 1994

    Google Scholar 

  25. Yamaguchi H, Miura, Ohsumi K, Ishimi N, Taguchi H, Ishiyama N, Shiraishi Y, Yamanoto T, Ogata S: Detection and characterization of antibodies to bacterial heat-shock protein 60 in sera of patients with primary biliary cirrhosis. Microbiol Immunol 38: 483-487, 1994

    Google Scholar 

  26. Fox JG, Dewhirst FE, Shen Z, Feng Y, Taylor NS, Paster BJ, Ericson RL, Lau CN, Correa P, Araya JC, Roa I: Hepatic helicobacter species identified in bile and gallbladder tissue from Chileans with chronic cholecystitis. Gastroenterology 114:755-763, 1998

    Google Scholar 

  27. Ward JM, Benveniste RE, Fox CH, Battles JK, Gonda MA, Tully JG: Autoimmunity in chronic active Helicobacter hepatitis of mice. Serum antibodies and expression of heat shock protein 70 in liver. Am J Pathol 148:509-517, 1996

    Google Scholar 

  28. Fox JG, Shen Z, Feng F, Dufover J, Kaplan MM, Dewhirst FE: Hepatobiliary Helicobacter spp. identified from patients with primary sclerosing cholangitis. Gastroenterology 114:A808, 1998

    Google Scholar 

  29. O'Donohue J, Fidler H, Garcia-Barcelo M, Nouri-Aria K, Williams R, McFadden J: Mycobacterial DNA not detected in liver sections from patients with primary biliary cirrhosis. J Hepatol 28:433-438, 1998

    Google Scholar 

  30. Tanaka A, Prindiville TP, Gish R, Solnick JV, Coppel RL, Keeffe EB, Ansari A, Gershwin ME: Are infectious agents involved in primary biliary cirrhosis? A PCR approach. J Hepatol 31:664-671, 1999

    Google Scholar 

  31. Hiramatsu K, Harada K, Tsuneyama, Sasaki M, Fujita S, Hashimoto T, Kaneko S, Kobayashi K, Nakanuma Y. Amplification and sequence analysis of partial bacterial 16S ribosomal RNA gene in gallbladder bile from patients with primary biliary cirrhosis. J Hepatol 33:9-18, 2000

    Google Scholar 

  32. Kreisel W, Siegel A, Bahler A, Spamer C, Schiltz E, Kist M, Seilnacht G, Klein R, Berg PA, Heilmann C: High prevalence of antibodies to calreticulin of the IgA class in primary biliary cirrhosis: A possible role of gut-derived bacterial antigens in its aetiology? Scand J Gastroenterol 34:623-628, 1999

    Google Scholar 

  33. Tanaka A, Nalbandian G, Leung PSC, Benson GD, Munoz S, Findor JA, Branch AD, Coppel RL, Ansari AA, Gershwin ME: Mucosal immunity and primary biliary cirrhosis: Presence of antimitochondrial antibodies in urine. Hepatology 32:910-915, 2000

    Google Scholar 

  34. Spangenberg HC, Deibert P, Kist M, Klein R, Berg PA, Blum HE, Kreisel W: Chronic liver disease with autoimmune phenomena triggered by Yersinia enterocolitica. Gastroenterology 116:L0439, 1999

    Google Scholar 

  35. Heathcote Y: The clinical expression of primary biliary cirrhosis. Semin Liver Dis 17:23-33, 1996

    Google Scholar 

  36. International Autoimmune Hepatitis Group report: Review of criteria for diagnosis of autoimmune hepatitis. J Hepatol 31:929-938, 1999

    Google Scholar 

  37. Kist M: Yersiniosis. In Clinical Laboratory Diagnostics: Use and Assessment of Clinical Laboratory Results. L Thomas, JT Whiocher (eds). Frankfurt/Main, Germany: TH Books Verlagsgesellschaft mbH, 1998, pp 1213-1217

    Google Scholar 

  38. Fournier PE, Marrie TJ, Raoult D: Diagnosis of Q-fever. J Clin Microbiol 36:1823-1834, 1998

    Google Scholar 

  39. Hagen EC, Ballieux BEBP, van Es LA, Daha MR, van der Woude FJ: Antineutrophile cytoplasmatic autoantibodies: a review of the antigens involved, the assays, and the clinical and possible pathogenetic consequences. Blood 81:1996-2002, 1993

    Google Scholar 

  40. Tan EM, Cohen AS, Fries JF, Masi AT, McShane DJ, Rothfield NF, Schaller JG, Talal N, Winchester RJ: The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum 25:1271-1277, 1982

    Google Scholar 

  41. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193:265-275, 1951

    Google Scholar 

  42. Laemmli UK: Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227:680-685, 1970

    Google Scholar 

  43. Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamid gels to nitrocellulose sheets: procedure and some applications, Proc Natl Acad Sci USA 76:4350-4354, 1979

    Google Scholar 

  44. Sternbach H, Engelhard R, Lezius AG: Rapid Isolation of highly active RNA polymerase from Escherichia coli and its subunits by matrix-bound heparin. Eur J Biochem 60:51-55, 1975

    Google Scholar 

  45. Ovchinnikov YA, Monastyrskaya GS, Gubanov VV, Guryev SO, Salomatina IS, Shuvaeva TM, Lipkin VM, Everdlov ED: The primary structure of E. coli RNA polymerase, nucleotide sequence of the rpoC gene and amino acid sequence of the beta'-subunit. Nucleic Acids Res 10:4035-4044, 1982

    Google Scholar 

  46. Cramer P, Bushnell DA, Fu J, Gnatt A, Maier Davis B, Thompson N, Burgess R, Edwards A, David P, Kornberg R: Architecture of RNA polymerase II and implications for the transcription mechanism. Science 288:640-649, 2000

    Google Scholar 

  47. Shigematsu H, Shimoda S, Nakamura M, Matsushita S, Nishimura Y, Sakamoto N, Ichiki Y, Niho Y, Gershwin ME, Ishibashi H. Fine specificity of T cells reactive to human PDC-E2 163-176 peptide, the immunodominant autoantigen in primary biliary cirrhosis: Implications for molecular mimicry and cross-recognition among mitochondrial autoantigens. Hepatology 32:901-909, 2000

    Google Scholar 

  48. Swidsinski A, Ladhoff A, Pernthaler A, Swidsinski S, Loening-Baucke S, Ortner M, Weber J, Hoffmann U, Schreiber S, Dietel M, Lochs H: Mucosal flora in inflammatory bowel disease. Gastroenterology 122:44-54, 2002

    Google Scholar 

  49. Grundmann H. Malaria: Mechanisms of immunity and immunization. Immun Infekt 20:156-160, 1992

    Google Scholar 

  50. Bogdanos DP, Choudhuri K, Vergani D: Molecular mimicry and autoimmune liver disease: virtuous intentions, malign consequences. Liver 21:225-232, 2001

    Google Scholar 

  51. Davidson A, Diamond B: Autoimmune diseases. N Engl J Med 345:340-350, 2001

    Google Scholar 

  52. Murakami M, Honjo T: Involvement of B-1 cells in mucosal immunity and autoimmunity. Immunol Today 16:534-539, 1995

    Google Scholar 

  53. Hayakawa K, Asano M, Shinton SA, Gui M, Allman D, Stewart CL, Silver J, Hardy RR: Positive selection of natural autoreactive B cells. Science 285:113-116, 1999

    Google Scholar 

  54. Jones DB, Coulson AFW, Duff GW: Sequence homologies between hsp60 and autoantigens. Immunol Today 14:115-118, 1993

    Google Scholar 

  55. Winrow V, Bird GL, Koskinas J, Blake, D, Williams R, Alexander GJM: Circulating IgA antibody against a 65-kDa heat shock protein in acute alcoholic hepatitis. J Hepatol 20:359-363, 1994

    Google Scholar 

  56. Anderton SM, van der Zee R, Goodacre JA: Inflammation activates self hsp60-specific T cells. Eur J Immunol 23:33-38, 1993

    Google Scholar 

  57. Lo WF, Woods AS, De Cloux A, Cotter RJ, Metcalf ES, Soloski MJ. Molecular mimicry mediated by MHC class Ib molecules after infection with Gram-negative pathogens. Nature Medicine 6:215-218, 2000

    Google Scholar 

Download references

Authors
  1. Kai-Wolfgang Roesler
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wolfgang Schmider
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Manfred Kist
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Stephen Batsford
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Emile Schiltz
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mathias Oelke
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Anja Tuczek
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Therese Dettenborn
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Dirk Behringer
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Wolfgang Kreisel
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Roesler, KW., Schmider, W., Kist, M. et al. Identification of β-Subunit of Bacterial RNA-Polymerase—A Non-Species-Specific Bacterial Protein—As Target of Antibodies in Primary Biliary Cirrhosis. Dig Dis Sci 48, 561–569 (2003). https://doi.org/10.1023/A:1022501102877

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1022501102877

Search

Navigation