Streptococcal Cell Wall Arthritis

Ronald L. Wilder1

1 National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, Maryland
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 15.10
DOI:  10.1002/0471142735.im1510s26
Online Posting Date:  May, 2001
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Abstract

Streptococcal cell wall (SCW) arthritis in rats is an experimentally‐induced inflammatory model with many features that resemble rheumatoid arthritis (RA) in humans. In this unit, Lewis rats are injected with an aqueous suspension of Group A SCW streptococcal cell wall peptidoglycan‐polysaccharide polymers (SCW PG‐PS) and observed for the development of arthritis. The resulting arthritis is biphasic. An acute phase typically develops within 48 hr, followed 10 to 21 days later by a chronic phase which persists for months. Support protocols are included for preparing Group A PG‐PS and measuring PG‐PS concentration.

     
 
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Table of Contents

  • Basic Protocol 1: Induction and Clinical Evaluation of Arthritis
  • Support Protocol 1: Preparation of Group a SCW 10S PG‐PS
  • Support Protocol 2: Measurement of Rhamnose Concentration to Estimate PG‐PS Concentration
  • Commentary
  • Tables
     
 
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Materials

Basic Protocol 1: Induction and Clinical Evaluation of Arthritis

  Materials
  • Lewis rats, female, 6‐ to 12‐weeks‐old or 100 to 200 g, specific‐pathogen‐free (Harlan Bioproducts)
  • 3 to 6 mg rhamnose/ml 10S PG‐PS suspension in sterile 0.85% NaCl (see protocol 2; alternatively, purchase from Lee Laboratories)
  • 0.85% NaCl, sterile
  • Scale appropriate for weighing rats
  • Calipers for measuring ankle diameter (optional)
  • Sonicator: Branson Sonifier 350 (VWR) or equivalent
  • 1‐ml syringes and 23‐G needles
  • Additional reagents and equipment for animal handling and restraint (unit 1.3), animal identification (unit 1.5), and intraperitoneal injection of rats (unit 1.6)

Support Protocol 1: Preparation of Group a SCW 10S PG‐PS

  Materials
  • Streptococcus pyogenes, Group A, D58 strain, (ATCC #43202)
  • Todd Hewett broth (Difco), pH 6.8 to 7.0, supplemented with 3 g dextrose/liter, autoclaved
  • 0.006 M PBS, pH 7.0: dilute PBS, pH 7.0 ( appendix 2A) 1/25 with H 2O (final NaCl concentration 0.006 M)
  • Dry ice/ethanol bath
  • 2% (w/v) SDS in 0.006 M PBS, pH 7.0
  • 2 M potassium chloride
  • 0.85% NaCl
  • Appropriately sized bacterial culture tubes and flasks
  • Blood agar plates (Difco)
  • 60°C water bath
  • Sonicator: Branson Sonifier 350 (VWR) or equivalent
  • Mid‐speed centrifuge and rotors (e.g., Sorvall, RC‐5B with SS‐34 or GSA rotors)
  • 0.45‐µm filters
  • Additional reagents and equipment for measuring rhamnose concentrations in cell wall pellet (see protocol 3)
CAUTION: Since Group A streptococci are human pathogens, precautions must be taken to avoid infecting oneself or others. Perform all sonications in a containment cabinet to prevent inhalation of aerosolized bacterial products.

Support Protocol 2: Measurement of Rhamnose Concentration to Estimate PG‐PS Concentration

  Materials
  • Rhamnose
  • PG‐PS suspensions for analysis (see protocol 2)
  • 6 vol concentrated sulfuric acid diluted with 1 vol H 2O (prepare fresh)
  • 3% (w/v) cysteine hydrochloride in H 2O
  • 16 × 150–mm test tubes
  • Boiling water bath
  • Spectrophotometer
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Literature Cited

Literature Cited
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   Lafyatis, R., Thompson, N.L., Remmers, E., Flanders, K., Roche, N.S., Kim, S.‐J., Case, J.P., Sporn, M.B., Roberts, A.B., and Wilder, R.L. 1989. Transforming growth factor–beta production by synovial tissues from rheumatoid patients and streptococcal cell wall arthritic rats: Studies on secretion by synovial fibroblast‐like cells and immunohistologic localization. J. Immunol. 143:1142‐1148.
   Lehman, T.J.A., Allen, J.B., Plotz, P., and Wilder, R.L. 1983. Polyarthritis in rats following systemic administration of cell walls from Lactobacillus casei in aqueous solution. Arthritis Rheum. 26:1259‐1265.
   McCartney‐Francis, N., Allen, J.B., Mizel, D.E., Albina, J.E., Xie, Q.‐W., Nathan, C.F., and Wahl, S.M. 1993. Suppression of arthritis by an inhibitor of nitric oxide synthase. J. Exp. Med. 178:749‐754.
   Sano, H., Forough, R., Maier, J.A.M., Case, J.P., Jackson, A., Engleka, K., Maciag, T., and Wilder, R.L. 1990. Detection of high levels of heparin binding growth factor‐1 (acidic FGF) in inflammatory arthritic joints. J. Cell Biol. 110:1417‐1426.
   Sano, H., Maciag, T., Maier, J.A.M., Crofford, L.J., Case, J.P., Maciag, T., and Wilder, R.L. 1992. In vivo cyclooxygenase expression in synovial tissues of patients with rheumatoid arthritis and osteoarthritis and rats with adjuvant and streptococcal cell wall arthritis. J. Clin. Invest. 89:97‐108.
   Sano, H., Engleka, K., Mathern, P., Hla, T., Crofford, L.J., Remmers, E.F., Jelsema, C., Goldmuntz, E., Maciag, T., and Wilder, R.L. 1993. Coexpression of tyrosine phosphoproteins, PDGF‐B and HBGF‐1 in situ in the synovial tissues of patients with rheumatoid arthritis and rats with adjuvant and streptococcal cell wall arthritis. J. Clin. Invest. 91:553‐565.
   Schleifer, K.H. and Kandler, O. 1972. Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bact. Rev. 36:407‐477.
   Schwab, J.H. and Smialowicz, R. 1975. Interaction of bacterial cell wall polymers and rat macrophages. Z. Immunitatsforsch. Exp. Klin. Immunol. 149:283‐288.
   Schwab, J.H., Allen, J.B., Anderle, S.K., Dalldorf, F., Eisenberg, R., and Cromartie, W.J. 1982. Relationship of complement to experimental arthritis induced in rats with streptococcal cell walls. Immunology 46:83‐88.
   Severijnen, A.J., Hazenberg, M.P., and Van de Merwe, J.P. 1988. Induction of chronic arthritis in rats by cell wall fragments of anaerobic coccoid rods isolated from the faecal flora of patients with Crohn's disease. Digestion 39:118‐125.
   Severijnen, A.J., Van Kleef, R., Hazenberg, M.P., and Van De Merwe, J.P. 1990. Chronic arthritis induced in rats by cell wall fragments of Eubacterium species from the human intestinal flora. Infect. Immun. 58:523‐528.
   Stein, H., Yarum, R., Levine, S., Dishon, T., and Ginsburg, I. 1973. Chronic self‐perpetuating arthritis induced in rabbits by a cell‐free extract of Group A streptoccoci. Proc. Soc. Exp. Biol. Med. 143:1106‐1112.
   Sternberg, E.M., Hill, J.M., Chrousos, G.P., Kamilaris, T., Listwak, S.J., Gold, P.W., and Wilder, R.L. 1989a. Inflammatory mediator‐induced hypothalamic‐pituitary‐adrenal axis activation is defective in streptococcal cell arthritis–susceptible Lewis rats. Proc. Natl. Acad. Sci. U.S.A. 86:2374‐2378.
   Sternberg, E.M., Young, W.S. III, Bernardini, R., Calogero, A.E., Chrousos, G.P., Gold, P.W., and Wilder, R.L. 1989b. A central nervous system defect in corticotropin‐releasing hormone biosynthesis is associated with susceptibility to streptococcal cell wall arthritis in Lewis rats. Proc. Natl. Acad. U.S.A. 86:4771‐4775.
   Stimpson, S.A., Brown, R.R., Anderle, S.K., Klapper, K.G., Clark, W.J., Cromartie, W.J., and Schwab, J.H. 1986. Arthropathic properties of cell wall polymers from normal flora bacteria. Infect. Immun. 51:240‐249.
   Taylor, J.E., Ross, D.A., and Goodacre, J.A. 1994. Group A streptococcal antigens and superantigens in the pathogenesis of autoimmune arthritis. Eur. J. Clin. Invest. 24:511‐521.
   Van De Langerijt, A.G.M., Kingston, A.E., Van Lent, P.L.E.M., Billingham, M.E.J., and Van Den Berg, W.B. 1994. Cross‐reactivity to proteoglycans in bacterial arthritis: Lack of evidence for in vivo role in induction of disease. Clin. Immunol. Immunopathol. 71:273‐280.
   Van Den Broek, M.F., Van Bruggen, M.C., Koopman, J.P., and Van Den Berg, W.B. 1992a. Gut flora induces and maintains resistance against streptococcal cell wall induced arthritis in F344 rats. Clin. Exp. Immunol. 88:313‐317.
   Van den Broek, M.F., Van De Langerijt, L.G.M., Van Bruggen, M.C.J., Billingham, M.E.J., and Van Den Berg, W.B. 1992b. Treatment of rats with monoclonal anti‐CD4 induces long‐term resistance to streptococcal cell wall–induced arthritis. Eur. J. Immunol. 22:57‐61.
   Wahl, S.M., Hunt, D.A., Allen, J.B., Wilder, R.L., Paglia, L., and Hunt, A.R. 1986. Bacterial cell wall‐induced hepatic granulomas: An in vivo model of T cell dependent fibrosis. J. Exp. Med. 163:884‐902.
   Wahl, S.M., Allen, J.B., Hines, K.L., Imamichi, T., Wahl, L.M., Furcht, L.T., and McCarthy, J.B. 1994. Synthetic fibronectin peptides suppress arthritis in rats by interrupting leukocyte adhesion and recruitment. J. Clin. Invest. 94:655‐662.
   Wilder, R.L., Calandra, G.B., Garvin, J., Hansen, C., and Wright, K. 1982. Strain and sex variation in the susceptibility to streptococcal cell wall induced polyarthritis in the rat. Arthritis Rheum. 25:1064‐1072.
   Wilder, R.L., Allen, J.B., and Hansen, C. 1987. Thymus‐dependent and ‐independent regulation of Ia antigen expression in situ by cells in the synovium of rats with streptococcal cell wall–induced arthritis: Differences in site and intensity of expression in euthymic, athymic and cyclosporin‐a‐treated LEW and F344 rats. J. Clin. Invest. 79:1160‐1171.
   Yocum, D.E., Allen, J.B., Wahl, S.M., and Wilder, R.L. 1986. Inhibition by cyclosporin of a streptococcal cell wall induced arthritis and hepatic granuloma formation in rats. Arthritis Rheum. 29:262‐273.
   Yocum, D.E., Lafyatis, R., Remmers, E., Schumacher, H.R., and Wilder, R.L. 1988. Hyperplastic synoviocytes from rats with streptococcal cell wall arthritis exhibit a transformed phenotype that is thymus‐dependent and retinoid inhibitable. Amer. J. Pathol. 132:38‐48.
   Yoshino, S., Cleland, L.G., Mayrhofer, G., Brown, R.R., and Schwab, J.H. 1991. Prevention of chronic erosive streptococcal cell wall‐induced arthritis in rats by treatment with a monoclonal antibody against the T cell antigen receptor αβ. J. Immunol. 146:4187‐4189.
Key Reference
   Crofford, L.J. and Wilder, R.L. 1996. Arthritis and autoimmunity in animals. In Arthritis and Allied Conditions (W.J. Koopman, ed.) pp. 565‐583. Lea & Febiger, Philadelphia.
  Contains an updated summary of various animal models of arthritis.
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