Skip to main content
NCBI home page
Annals of the Rheumatic Diseases logoLink to Annals of the Rheumatic Diseases
. 2001 Mar;60(3):269–274. doi: 10.1136/ard.60.3.269

Characterisation of Eubacterium cell wall: peptidoglycan structure determines arthritogenicity

X Zhang 1, M Rimpilainen 1, P Toivanen 1
PMCID: PMC1753583  PMID: 11171690

Abstract

OBJECTIVE—To elucidate factors involved in the arthritogenicity of bacterial cell walls.
METHODS—For characterisation of an arthritogenic Eubacterium aerofaciens cell wall, peptidoglycan-polysaccharide (PG-PS) polymers were isolated by removing cell wall associated proteins (CWPs), PG and PS moieties were separated, and an attempt was made to de-O-acetylate PG-PS. The cell wall of E limosum was used as a non-arthritogenic control. The chemical composition of these cell wall preparations was analysed by gas chromatography-mass spectrometry. Also, their ability to resist lysozyme degradation and to sustain experimental chronic arthritis was tested.
RESULTS—The observations made with the cell wall of E aerofaciens, an anaerobic habitant of the human intestine, were compared with those reported from a pathogenic Streptococcus, showing that in both strains a complex consisting of PG-PS is required for the induction of chronic arthritis. The PS moiety most probably protects PG from enzyme degradation, allowing prolonged tissue persistence and leading to the chronic synovial inflammation. CWPs attached to PG-PS are not necessary for this function. O-Acetylation of PG, which is required for arthritogenicity of the streptococcal cell wall, seems not to be present in the arthritogenic E aerofaciens PG or only occurs to a small degree; attempts to de-O-acylate the E aerofaciens cell wall did not affect its arthritogenicity or lysozyme resistance.
CONCLUSION—The results obtained indicate that the source of bacterial cell wall plays no part in the chemical or structural requirements for PG to induce chronic cell wall arthritis in the rats; the chemical structure of the PG moiety is decisive.



Full Text

The Full Text of this article is available as a PDF (174.8 KB).

Figure 1  .

Figure 1  

Open in a new tab

Gram positive bacterial cell wall contains mainly peptidoglycan (PG), polysaccharide (PS), and cell wall associated proteins (CWPs). The CWPs outside the PS and PG layers can be removed by proteolytic enzymes, whereas the proteins inside the PG layer are difficult to remove without breaking the PG structure. PS binds covalently to the PG layer, and protects PG polymers from the degrading enzymes. PG consists of several layers (up to 70) of sugar chains composed of N-acetylglucosamine and N-acetylmuramic acid alternately linked each other. Short peptides bound to the N-acetylmuramic acid (muramyl peptides) connect the sugar chains. PG and the muramyl peptides possess multiple biological activities.

Figure 2  .

Figure 2  

Open in a new tab

(A) A single intraperitoneal (IP) injection of E aerofaciens peptidoglycan-polysaccharide (PG-PS; 150 µg/g body weight of rat) induced a more severe chronic arthritis than that induced with a crude cell wall (140 µg/g body weight of rat) (p<0.02 by non-parametric Mann-Whitney U test). Each black symbol represents a mean (SEM) of six rats. Each white symbol represents a mean (SEM) of 11 rats. (B) Rats injected with E limosum PG-PS (200 µg/g body weight of rat) IP or with the crude cell wall (200 µg/g body weight of rat) developed only mild acute arthritis. Each black symbol represents a mean (SEM) of eight rats. Each white symbol represents a mean (SEM) of six to 12 rats.

Figure 3  .

Figure 3  

Open in a new tab

(A) Rats injected with E aerofaciens peptidoglycan (PG; 33 µg/g body weight of rat) intraperitoneally (IP) deprived of polysaccharide (PS), developed only acute arthritis. Rats injected with E aerofaciens PS alone (122 µg/g body weight of rat) did not develop any arthritis. Each symbol represents a mean (SEM) of five rats. (B) Rats injected with E aerofaciens de-O-acylated cell wall (140 µg/g body weight of rat) IP developed arthritis similar to that seen after injection of the crude cell wall (140 µg/g body weight of rat). Each black symbol represents a mean (SEM) of six rats. Each white symbol represents a mean (SEM) of 11 rats.    

Selected References

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

  1. 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]
  2. Barnes P. F., Mehra V., Hirschfield G. R., Fong S. J., Abou-Zeid C., Rook G. A., Hunter S. W., Brennan P. J., Modlin R. L. Characterization of T cell antigens associated with the cell wall protein-peptidoglycan complex of Mycobacterium tuberculosis. J Immunol. 1989 Oct 15;143(8):2656–2662. [PubMed] [Google Scholar]
  3. Chetty C., Brown R. R., Schwab J. H. Edema-producing activity of group A streptococcal polysaccharide and its possible role in the pathogenesis of cell wall-induced polyarthritis. J Exp Med. 1983 Apr 1;157(4):1089–1100. doi: 10.1084/jem.157.4.1089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clarke A. J., Dupont C. O-acetylated peptidoglycan: its occurrence, pathobiological significance, and biosynthesis. Can J Microbiol. 1992 Feb;38(2):85–91. doi: 10.1139/m92-014. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. DeJoy S. Q., Ferguson-Chanowitz K. M., Sapp T. M., Oronsky A. L., Lapierre L. A., Zabriskie J. B., Kerwar S. S. M protein deficient streptococcal cell walls can induce acute and chronic arthritis rats. Cell Immunol. 1990 Feb;125(2):526–534. doi: 10.1016/0008-8749(90)90105-z. [DOI] [PubMed] [Google Scholar]
  7. Doyle R. J., Streips U. N., Fan V. S., Brown W. C., Mobley H., Mansfield J. M. Cell wall protein in Bacillus subtilis. J Bacteriol. 1977 Jan;129(1):547–549. doi: 10.1128/jb.129.1.547-549.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Eerola E., Möttönen T., Hannonen P., Luukkainen R., Kantola I., Vuori K., Tuominen J., Toivanen P. Intestinal flora in early rheumatoid arthritis. Br J Rheumatol. 1994 Nov;33(11):1030–1038. doi: 10.1093/rheumatology/33.11.1030. [DOI] [PubMed] [Google Scholar]
  9. Fleming T. J., Wallsmith D. E., Rosenthal R. S. Arthropathic properties of gonococcal peptidoglycan fragments: implications for the pathogenesis of disseminated gonococcal disease. Infect Immun. 1986 May;52(2):600–608. doi: 10.1128/iai.52.2.600-608.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Foster T. J., McDevitt D. Surface-associated proteins of Staphylococcus aureus: their possible roles in virulence. FEMS Microbiol Lett. 1994 May 15;118(3):199–205. doi: 10.1111/j.1574-6968.1994.tb06828.x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Gilbart J., Harrison J., Parks C., Fox A. Analysis of the amino acid and sugar composition of streptococcal cell walls by gas chromatography-mass spectrometry. J Chromatogr. 1988 Jun 10;441(2):323–333. doi: 10.1016/s0021-9673(01)83875-9. [DOI] [PubMed] [Google Scholar]
  13. Hill S. A., Judd R. C. Identification and characterization of peptidoglycan-associated proteins in Neisseria gonorrhoeae. Infect Immun. 1989 Nov;57(11):3612–3618. doi: 10.1128/iai.57.11.3612-3618.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hoijer M. A., Melief M. J., Calafat J., Roos D., van den Beemd R. W., van Dongen J. J., Hazenberg M. P. Expression and intracellular localization of the human N-acetylmuramyl-L-alanine amidase, a bacterial cell wall-degrading enzyme. Blood. 1997 Aug 1;90(3):1246–1254. [PubMed] [Google Scholar]
  15. Jalava J., Mäntymaa M. L., Ekblad U., Toivanen P., Skurnik M., Lassila O., Alanen A. Bacterial 16S rDNA polymerase chain reaction in the detection of intra-amniotic infection. Br J Obstet Gynaecol. 1996 Jul;103(7):664–669. doi: 10.1111/j.1471-0528.1996.tb09835.x. [DOI] [PubMed] [Google Scholar]
  16. Lehman T. J., Allen J. B., Plotz P. H., Wilder R. L. Bacterial cell wall composition, lysozyme resistance, and the induction of chronic arthritis in rats. Rheumatol Int. 1985;5(4):163–167. doi: 10.1007/BF00541517. [DOI] [PubMed] [Google Scholar]
  17. Majcherczyk P. A., Langen H., Heumann D., Fountoulakis M., Glauser M. P., Moreillon P. Digestion of Streptococcus pneumoniae cell walls with its major peptidoglycan hydrolase releases branched stem peptides carrying proinflammatory activity. J Biol Chem. 1999 Apr 30;274(18):12537–12543. doi: 10.1074/jbc.274.18.12537. [DOI] [PubMed] [Google Scholar]
  18. Melancon-Kaplan J., Hunter S. W., McNeil M., Stewart C., Modlin R. L., Rea T. H., Convit J., Salgame P., Mehra V., Bloom B. R. Immunological significance of Mycobacterium leprae cell walls. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1917–1921. doi: 10.1073/pnas.85.6.1917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pancholi V., Fischetti V. A. Isolation and characterization of the cell-associated region of group A streptococcal M6 protein. J Bacteriol. 1988 Jun;170(6):2618–2624. doi: 10.1128/jb.170.6.2618-2624.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Peltonen R., Nenonen M., Helve T., Hänninen O., Toivanen P., Eerola E. Faecal microbial flora and disease activity in rheumatoid arthritis during a vegan diet. Br J Rheumatol. 1997 Jan;36(1):64–68. doi: 10.1093/rheumatology/36.1.64. [DOI] [PubMed] [Google Scholar]
  21. Rosenthal R. S., Dziarski R. Isolation of peptidoglycan and soluble peptidoglycan fragments. Methods Enzymol. 1994;235:253–285. doi: 10.1016/0076-6879(94)35146-5. [DOI] [PubMed] [Google Scholar]
  22. Rosenthal R. S., Folkening W. J., Miller D. R., Swim S. C. Resistance of O-acetylated gonococcal peptidoglycan to human peptidoglycan-degrading enzymes. Infect Immun. 1983 Jun;40(3):903–911. doi: 10.1128/iai.40.3.903-911.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sato K., Saito H., Tomioka H., Yokokura T. Enhancement of host resistance against Listeria infection by Lactobacillus casei: efficacy of cell wall preparation of Lactobacillus casei. Microbiol Immunol. 1988;32(12):1189–1200. doi: 10.1111/j.1348-0421.1988.tb01483.x. [DOI] [PubMed] [Google Scholar]
  24. Schwab J. H. Phlogistic properties of peptidoglycan-polysaccharide polymers from cell walls of pathogenic and normal-flora bacteria which colonize humans. Infect Immun. 1993 Nov;61(11):4535–4539. doi: 10.1128/iai.61.11.4535-4539.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Severijnen A. J., van Kleef R., Hazenberg M. P., van de Merwe J. P. Cell wall fragments from major residents of the human intestinal flora induce chronic arthritis in rats. J Rheumatol. 1989 Aug;16(8):1061–1068. [PubMed] [Google Scholar]
  26. Severijnen A. J., van Kleef R., Hazenberg M. P., van de Merwe J. P. Chronic arthritis induced in rats by cell wall fragments of Eubacterium species from the human intestinal flora. Infect Immun. 1990 Feb;58(2):523–528. doi: 10.1128/iai.58.2.523-528.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Simelyte E., Rimpiläinen M., Lehtonen L., Zhang X., Toivanen P. Bacterial cell wall-induced arthritis: chemical composition and tissue distribution of four Lactobacillus strains. Infect Immun. 2000 Jun;68(6):3535–3540. doi: 10.1128/iai.68.6.3535-3540.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Simelyte E., Rimpiläinen M., Rantakokko K., Lehtonen L., Zhang X., Aho H., Isomäki P., Toivanen P. Tissue distribution and persistence of arthritogenic and non-arthritogenic Eubacterium cell walls. Clin Exp Rheumatol. 1999 May-Jun;17(3):281–288. [PubMed] [Google Scholar]
  29. Stimpson S. A., Brown R. R., Anderle S. K., Klapper D. G., Clark R. L., Cromartie W. J., Schwab J. H. Arthropathic properties of cell wall polymers from normal flora bacteria. Infect Immun. 1986 Jan;51(1):240–249. doi: 10.1128/iai.51.1.240-249.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Stimpson S. A., Lerch R. A., Cleland D. R., Yarnall D. P., Clark R. L., Cromartie W. J., Schwab J. H. Effect of acetylation on arthropathic activity of group A streptococcal peptidoglycan-polysaccharide fragments. Infect Immun. 1987 Jan;55(1):16–23. doi: 10.1128/iai.55.1.16-23.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Striker R., Kline M. E., Haak R. A., Rest R. F., Rosenthal R. S. Degradation of gonococcal peptidoglycan by granule extract from human neutrophils: demonstration of N-acetylglucosaminidase activity that utilizes peptidoglycan substrates. Infect Immun. 1987 Nov;55(11):2579–2584. doi: 10.1128/iai.55.11.2579-2584.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tomai M., Kotb M., Majumdar G., Beachey E. H. Superantigenicity of streptococcal M protein. J Exp Med. 1990 Jul 1;172(1):359–362. doi: 10.1084/jem.172.1.359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ton-That H., Faull K. F., Schneewind O. Anchor structure of staphylococcal surface proteins. A branched peptide that links the carboxyl terminus of proteins to the cell wall. J Biol Chem. 1997 Aug 29;272(35):22285–22292. doi: 10.1074/jbc.272.35.22285. [DOI] [PubMed] [Google Scholar]
  34. Zhang X., Rimpiläinen M., Simelyte E., Toivanen P. What determines arthritogenicity of bacterial cell wall? A study on Eubacterium cell wall-induced arthritis. Rheumatology (Oxford) 2000 Mar;39(3):274–282. doi: 10.1093/rheumatology/39.3.274. [DOI] [PubMed] [Google Scholar]
  35. Zhang Y., Gripenberg-Lerche C., Söderström K. O., Toivanen A., Toivanen P. Antibiotic prophylaxis and treatment of reactive arthritis. Lessons from an animal model. Arthritis Rheum. 1996 Jul;39(7):1238–1243. doi: 10.1002/art.1780390725. [DOI] [PubMed] [Google Scholar]
  36. de Maagd R. A., Wientjes F. B., Lugtenberg B. J. Evidence for divalent cation (Ca2+)-stabilized oligomeric proteins and covalently bound protein-peptidoglycan complexes in the outer membrane of Rhizobium leguminosarum. J Bacteriol. 1989 Jul;171(7):3989–3995. doi: 10.1128/jb.171.7.3989-3995.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. van der Heijden I. M., Wilbrink B., Tchetverikov I., Schrijver I. A., Schouls L. M., Hazenberg M. P., Breedveld F. C., Tak P. P. Presence of bacterial DNA and bacterial peptidoglycans in joints of patients with rheumatoid arthritis and other arthritides. Arthritis Rheum. 2000 Mar;43(3):593–598. doi: 10.1002/1529-0131(200003)43:3<593::AID-ANR16>3.0.CO;2-1. [DOI] [PubMed] [Google Scholar]

Articles from Annals of the Rheumatic Diseases are provided here courtesy of BMJ Publishing Group

RESOURCES