Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1996 Mar;64(3):818–824. doi: 10.1128/iai.64.3.818-824.1996

Anti-Hsp65 antibodies recognize M proteins of group A streptococci.

A Quinn 1, T M Shinnick 1, M W Cunningham 1
PMCID: PMC173842  PMID: 8641786

Abstract

Group A streptococcal M protein and the mycobacterial heat shock protein, hsp65, are strong bacterial immunogens that have been linked to arthritis and autoimmunity. Recent evidence has shown that streptococcal arthritis and adjuvant arthritis may be related to epitopes shared between group A streptococci and hsp65. We investigated the possibility that immunological similarities were shared between streptococcal M protein and hsp65. Antibodies against the 65-kDa heat shock protein of Mycobacterium tuberculosis were tested for reactivity with group A streptococci and purified recombinant M proteins (rM5 and rM6). Rabbit polyclonal anti-hsp65 serum was highly reactive with M type 5 Streptococcus pyogenes and rM5 and rM6 proteins in an enzyme-linked immunosorbent assay (ELISA). A mouse anti-hsp65 monoclonal antibody (MAb), IIC8, reacted with streptococcal M types 5, 6, 19, 24, and 49 in an ELISA but showed no reactivity with an isogenic streptococcal mutant which did not express M protein. Anti-hsp65 MAb IIC8 recognized rM5 and rM6 proteins in the ELISA, and MAbs IIC8 and IIH9 reacted strongly with rM6 protein in Western immunoblots. The binding of M protein by anti-hsp65 MAbs was shown to be inhibited by both hsp65 and M protein. These data show that anti-hsp65 antibodies recognize streptococcal M proteins.

Full Text

The Full Text of this article is available as a PDF (288.8 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson D. C., Barry M. E., Buchanan T. M. Exact definition of species-specific and cross-reactive epitopes of the 65-kilodalton protein of Mycobacterium leprae using synthetic peptides. J Immunol. 1988 Jul 15;141(2):607–613. [PubMed] [Google Scholar]
  2. Baird R. W., Bronze M. S., Kraus W., Hill H. R., Veasey L. G., Dale J. B. Epitopes of group A streptococcal M protein shared with antigens of articular cartilage and synovium. J Immunol. 1991 May 1;146(9):3132–3137. [PubMed] [Google Scholar]
  3. Beachey E. H., Campbell G. L., Ofek I. Peptic digestion of streptococcal M protein. II. Extraction of M antigen from group A streptococci with pepsin. Infect Immun. 1974 May;9(5):891–896. doi: 10.1128/iai.9.5.891-896.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bessen D., Jones K. F., Fischetti V. A. Evidence for two distinct classes of streptococcal M protein and their relationship to rheumatic fever. J Exp Med. 1989 Jan 1;169(1):269–283. doi: 10.1084/jem.169.1.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Billingham M. E., Carney S., Butler R., Colston M. J. A mycobacterial 65-kD heat shock protein induces antigen-specific suppression of adjuvant arthritis, but is not itself arthritogenic. J Exp Med. 1990 Jan 1;171(1):339–344. doi: 10.1084/jem.171.1.339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Buchanan T. M., Nomaguchi H., Anderson D. C., Young R. A., Gillis T. P., Britton W. J., Ivanyi J., Kolk A. H., Closs O., Bloom B. R. Characterization of antibody-reactive epitopes on the 65-kilodalton protein of Mycobacterium leprae. Infect Immun. 1987 Apr;55(4):1000–1003. doi: 10.1128/iai.55.4.1000-1003.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burnie J. P., Matthews R. C. Heat shock protein 88 and Aspergillus infection. J Clin Microbiol. 1991 Oct;29(10):2099–2106. doi: 10.1128/jcm.29.10.2099-2106.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen I. R. Autoimmunity to chaperonins in the pathogenesis of arthritis and diabetes. Annu Rev Immunol. 1991;9:567–589. doi: 10.1146/annurev.iy.09.040191.003031. [DOI] [PubMed] [Google Scholar]
  9. Cromartie W. J., Craddock J. G., Schwab J. H., Anderle S. K., Yang C. H. Arthritis in rats after systemic injection of streptococcal cells or cell walls. J Exp Med. 1977 Dec 1;146(6):1585–1602. doi: 10.1084/jem.146.6.1585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cunningham M. W., Antone S. M., Gulizia J. M., McManus B. M., Fischetti V. A., Gauntt C. J. Cytotoxic and viral neutralizing antibodies crossreact with streptococcal M protein, enteroviruses, and human cardiac myosin. Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1320–1324. doi: 10.1073/pnas.89.4.1320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cunningham M. W., Beachey E. H. Peptic digestion of streptococcal M protein. I. Effect of digestion at suboptimal pH upon the biological and immunochemical properties of purified M protein extracts. Infect Immun. 1974 Feb;9(2):244–248. doi: 10.1128/iai.9.2.244-248.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cunningham M. W., McCormack J. M., Talaber L. R., Harley J. B., Ayoub E. M., Muneer R. S., Chun L. T., Reddy D. V. Human monoclonal antibodies reactive with antigens of the group A Streptococcus and human heart. J Immunol. 1988 Oct 15;141(8):2760–2766. [PubMed] [Google Scholar]
  13. Cunningham M. W., Swerlick R. A. Polyspecificity of antistreptococcal murine monoclonal antibodies and their implications in autoimmunity. J Exp Med. 1986 Oct 1;164(4):998–1012. doi: 10.1084/jem.164.4.998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dale J. B., Beachey E. H. Epitopes of streptococcal M proteins shared with cardiac myosin. J Exp Med. 1985 Aug 1;162(2):583–591. doi: 10.1084/jem.162.2.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. DeJoy S. Q., Ferguson K. M., Sapp T. M., Zabriskie J. B., Oronsky A. L., Kerwar S. S. Streptococcal cell wall arthritis. Passive transfer of disease with a T cell line and crossreactivity of streptococcal cell wall antigens with Mycobacterium tuberculosis. J Exp Med. 1989 Aug 1;170(2):369–382. doi: 10.1084/jem.170.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Esaguy N., Aguas A. P., van Embden J. D., Silva M. T. Mycobacteria and human autoimmune disease: direct evidence of cross-reactivity between human lactoferrin and the 65-kilodalton protein of tubercle and leprosy bacilli. Infect Immun. 1991 Mar;59(3):1117–1125. doi: 10.1128/iai.59.3.1117-1125.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fischetti V. A. Streptococcal M protein: molecular design and biological behavior. Clin Microbiol Rev. 1989 Jul;2(3):285–314. doi: 10.1128/cmr.2.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fox A., Brown R. R., Anderle S. K., Chetty C., Cromartie W. J., Gooder H., Schwab J. H. Arthropathic properties related to the molecular weight of peptidoglycan-polysaccharide polymers of streptococcal cell walls. Infect Immun. 1982 Mar;35(3):1003–1010. doi: 10.1128/iai.35.3.1003-1010.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Fung J. C., Wicher K., McCarty M. Immunochemical analysis of streptococcal group A, B, and C carbohydrates, with emphasis on group A. Infect Immun. 1982 Jul;37(1):209–215. doi: 10.1128/iai.37.1.209-215.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gaston J. S., Life P. F., Jenner P. J., Colston M. J., Bacon P. A. Recognition of a mycobacteria-specific epitope in the 65-kD heat-shock protein by synovial fluid-derived T cell clones. J Exp Med. 1990 Mar 1;171(3):831–841. doi: 10.1084/jem.171.3.831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gillis T. P., Buchanan T. M. Production and partial characterization of monoclonal antibodies to Mycobacterium leprae. Infect Immun. 1982 Jul;37(1):172–178. doi: 10.1128/iai.37.1.172-178.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hedstrom R., Culpepper J., Harrison R. A., Agabian N., Newport G. A major immunogen in Schistosoma mansoni infections is homologous to the heat-shock protein Hsp70. J Exp Med. 1987 May 1;165(5):1430–1435. doi: 10.1084/jem.165.5.1430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Holmdahl R., Rubin K., Klareskog L., Larsson E., Wigzell H. Characterization of the antibody response in mice with type II collagen-induced arthritis, using monoclonal anti-type II collagen antibodies. Arthritis Rheum. 1986 Mar;29(3):400–410. doi: 10.1002/art.1780290314. [DOI] [PubMed] [Google Scholar]
  24. Holoshitz J., Klajman A., Drucker I., Lapidot Z., Yaretzky A., Frenkel A., van Eden W., Cohen I. R. T lymphocytes of rheumatoid arthritis patients show augmented reactivity to a fraction of mycobacteria cross-reactive with cartilage. Lancet. 1986 Aug 9;2(8502):305–309. doi: 10.1016/s0140-6736(86)90003-6. [DOI] [PubMed] [Google Scholar]
  25. Holoshitz J., Naparstek Y., Ben-Nun A., Cohen I. R. Lines of T lymphocytes induce or vaccinate against autoimmune arthritis. Science. 1983 Jan 7;219(4580):56–58. doi: 10.1126/science.6336851. [DOI] [PubMed] [Google Scholar]
  26. Jindal S., Dudani A. K., Singh B., Harley C. B., Gupta R. S. Primary structure of a human mitochondrial protein homologous to the bacterial and plant chaperonins and to the 65-kilodalton mycobacterial antigen. Mol Cell Biol. 1989 May;9(5):2279–2283. doi: 10.1128/mcb.9.5.2279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Jones D. B., Coulson A. F., Duff G. W. Sequence homologies between hsp60 and autoantigens. Immunol Today. 1993 Mar;14(3):115–118. doi: 10.1016/0167-5699(93)90210-C. [DOI] [PubMed] [Google Scholar]
  28. KAPLAN M. H. IMMUNOLOGIC RELATION OF STREPTOCOCCAL AND TISSUE ANTIGENS. I. PROPERTIES OF AN ANTIGEN IN CERTAIN STRAINS OF GROUP A STREPTOCOCCI EXHIBITING AN IMMUNOLOGIC CROSS-REACTION WITH HUMAN HEART TISSUE. J Immunol. 1963 Apr;90:595–606. [PubMed] [Google Scholar]
  29. Karlsson-Parra A., Söderström K., Ferm M., Ivanyi J., Kiessling R., Klareskog L. Presence of human 65 kD heat shock protein (hsp) in inflamed joints and subcutaneous nodules of RA patients. Scand J Immunol. 1990 Jun;31(6):283–288. doi: 10.1111/j.1365-3083.1990.tb02770.x. [DOI] [PubMed] [Google Scholar]
  30. Kaufmann S. H., Schoel B., van Embden J. D., Koga T., Wand-Württenberger A., Munk M. E., Steinhoff U. Heat-shock protein 60: implications for pathogenesis of and protection against bacterial infections. Immunol Rev. 1991 Jun;121:67–90. doi: 10.1111/j.1600-065x.1991.tb00823.x. [DOI] [PubMed] [Google Scholar]
  31. LANCEFIELD R. C. Current knowledge of type-specific M antigens of group A streptococci. J Immunol. 1962 Sep;89:307–313. [PubMed] [Google Scholar]
  32. Lehner T., Lavery E., Smith R., van der Zee R., Mizushima Y., Shinnick T. Association between the 65-kilodalton heat shock protein, Streptococcus sanguis, and the corresponding antibodies in Behçet's syndrome. Infect Immun. 1991 Apr;59(4):1434–1441. doi: 10.1128/iai.59.4.1434-1441.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Manjula B. N., Trus B. L., Fischetti V. A. Presence of two distinct regions in the coiled-coil structure of the streptococcal Pep M5 protein: relationship to mammalian coiled-coil proteins and implications to its biological properties. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1064–1068. doi: 10.1073/pnas.82.4.1064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Morrison R. P., Belland R. J., Lyng K., Caldwell H. D. Chlamydial disease pathogenesis. The 57-kD chlamydial hypersensitivity antigen is a stress response protein. J Exp Med. 1989 Oct 1;170(4):1271–1283. doi: 10.1084/jem.170.4.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Mouw A. R., Beachey E. H., Burdett V. Molecular evolution of streptococcal M protein: cloning and nucleotide sequence of the type 24 M protein gene and relation to other genes of Streptococcus pyogenes. J Bacteriol. 1988 Feb;170(2):676–684. doi: 10.1128/jb.170.2.676-684.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Phillips G. N., Jr, Flicker P. F., Cohen C., Manjula B. N., Fischetti V. A. Streptococcal M protein: alpha-helical coiled-coil structure and arrangement on the cell surface. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4689–4693. doi: 10.1073/pnas.78.8.4689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Plikaytis B. B., Carlone G. M., Pau C. P., Wilkinson H. W. Purified 60-kilodalton Legionella protein antigen with Legionella-specific and nonspecific epitopes. J Clin Microbiol. 1987 Nov;25(11):2080–2084. doi: 10.1128/jcm.25.11.2080-2084.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Res P. C., Schaar C. G., Breedveld F. C., van Eden W., van Embden J. D., Cohen I. R., de Vries R. R. Synovial fluid T cell reactivity against 65 kD heat shock protein of mycobacteria in early chronic arthritis. Lancet. 1988 Aug 27;2(8609):478–480. doi: 10.1016/s0140-6736(88)90123-7. [DOI] [PubMed] [Google Scholar]
  39. Shinnick T. M. Heat shock proteins as antigens of bacterial and parasitic pathogens. Curr Top Microbiol Immunol. 1991;167:145–160. doi: 10.1007/978-3-642-75875-1_9. [DOI] [PubMed] [Google Scholar]
  40. Shinnick T. M. The 65-kilodalton antigen of Mycobacterium tuberculosis. J Bacteriol. 1987 Mar;169(3):1080–1088. doi: 10.1128/jb.169.3.1080-1088.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Stuart J. M., Cremer M. A., Townes A. S., Kang A. H. Type II collagen-induced arthritis in rats. Passive transfer with serum and evidence that IgG anticollagen antibodies can cause arthritis. J Exp Med. 1982 Jan 1;155(1):1–16. doi: 10.1084/jem.155.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Stuart J. M., Dixon F. J. Serum transfer of collagen-induced arthritis in mice. J Exp Med. 1983 Aug 1;158(2):378–392. doi: 10.1084/jem.158.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Svartman M., Potter E. V., Poon-King T., Earle D. P. Immunoglobulins and complement components in synovial fluid of patients with acute rheumatic fever. J Clin Invest. 1975 Jul;56(1):111–117. doi: 10.1172/JCI108059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Terato K., Hasty K. A., Reife R. A., Cremer M. A., Kang A. H., Stuart J. M. Induction of arthritis with monoclonal antibodies to collagen. J Immunol. 1992 Apr 1;148(7):2103–2108. [PubMed] [Google Scholar]
  45. Trentham D. E., Dynesius R. A., David J. R. Passive transfer by cells of type II collagen-induced arthritis in rats. J Clin Invest. 1978 Aug;62(2):359–366. doi: 10.1172/JCI109136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Trentham D. E., Townes A. S., Kang A. H. Autoimmunity to type II collagen an experimental model of arthritis. J Exp Med. 1977 Sep 1;146(3):857–868. doi: 10.1084/jem.146.3.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Watson K. Microbial stress proteins. Adv Microb Physiol. 1990;31:183–223. doi: 10.1016/s0065-2911(08)60122-8. [DOI] [PubMed] [Google Scholar]
  48. Zhang Z. Y., Lee C. S., Lider O., Weiner H. L. Suppression of adjuvant arthritis in Lewis rats by oral administration of type II collagen. J Immunol. 1990 Oct 15;145(8):2489–2493. [PubMed] [Google Scholar]
  49. van Eden W. Heat-shock proteins as immunogenic bacterial antigens with the potential to induce and regulate autoimmune arthritis. Immunol Rev. 1991 Jun;121:5–28. doi: 10.1111/j.1600-065x.1991.tb00821.x. [DOI] [PubMed] [Google Scholar]
  50. van Eden W., Holoshitz J., Nevo Z., Frenkel A., Klajman A., Cohen I. R. Arthritis induced by a T-lymphocyte clone that responds to Mycobacterium tuberculosis and to cartilage proteoglycans. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5117–5120. doi: 10.1073/pnas.82.15.5117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. van Eden W., Thole J. E., van der Zee R., Noordzij A., van Embden J. D., Hensen E. J., Cohen I. R. Cloning of the mycobacterial epitope recognized by T lymphocytes in adjuvant arthritis. Nature. 1988 Jan 14;331(6152):171–173. doi: 10.1038/331171a0. [DOI] [PubMed] [Google Scholar]
  52. van den Broek M. F., Hogervorst E. J., Van Bruggen M. C., Van Eden W., van der Zee R., van den Berg W. B. Protection against streptococcal cell wall-induced arthritis by pretreatment with the 65-kD mycobacterial heat shock protein. J Exp Med. 1989 Aug 1;170(2):449–466. doi: 10.1084/jem.170.2.449. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES