Journal of Molecular Medicine

, Volume 83, Issue 7, pp 535–541

TLR-2 gene Arg753Gln polymorphism is strongly associated with acute rheumatic fever in children

  • Afig Berdeli
  • Handan Ak Celik
  • Ruhi Özyürek
  • Buket Dogrusoz
  • Hikmet Hakan Aydin
Rapid Communication


The recently described family of toll-like receptors (TLRs) is a key player in host immunity by mediating inflammatory reactions against a wide range of pathogens. Mutations and polymorphisms in TLRs have revealed the importance of TLRs in human defence against diseases. TLR-2 is reported to interact with different bacterial structures, including lipoproteins, peptidoglycan and lipoteichoic acid. To assess the role of TLR-2 gene polymorphism in acute rheumatic fever (ARF) etiopathology, 61 independent Caucasian Turkish patients and 91 child and 116 adult controls were studied. Antistreptolycin O, C-reactive protein, sedimentation and white blood cell counts were studied to evaluate the clinical characteristics of the patients. Genomic DNA was extracted from peripheral blood using a standard column extraction technique. The Arg753Gln and Arg677Trp polymorphisms were genotyped by polymerase chain reaction (PCR) restriction fragment length polymorphism. The PCR products for the TLR-2 gene were analysed on 1.5% agarose gel pre-stained with ethidium bromide. Compared with healthy adult controls, the Arg753Arg genotype was significantly decreased in the entire group of ARF cases [odds ratio (OR) 0.01, 95% confidence interval (95% CI) 0.0034–0.031, p<0.0001]. Significantly, ARF patients were just 16 times more frequent with Gln allele (OR 15.6, 95% CI 7.87–30.8, p<0.0001). Moreover, evidence for an intensifying effect of the Gln allele was noteworthy when patients with Arg753Gln genotype were compared with healthy controls (OR 97.1, 95% CI 32.5–290, p<0.0001). However, no Arg677Trp polymorphism was detected in either patients or controls. Our data suggest that there is strong evidence for the biological role of TLR-2 in ARF. The common TLR-2 Arg to Gln polymorphism at position 753 significantly contributes to the pathogenesis of ARF. These results will allow the construction of a profile of individuals prone to ARF and may assist in developing new therapies.


Acute rheumatic fever TLR-2 Gene polymorphism Single nucleotide polymorphism Infections 



Acute rheumatic fever


Rheumatic heart disease


Toll-like receptor


Polymerase chain reaction


Restriction fragment length polymorphism


Antistreptolysin O


C-reactive protein


Erythrocyte sedimentation rate


White blood cells count


Nuclear transcription factor kappa B


  1. 1.
    Kaplan E (1996) Recent epidemiology of group A streptococcal infections in North America and abroad: an overview. Pediatrics 97:S945–S948Google Scholar
  2. 2.
    Krishna Kumar R et al (1999) Epidemiology of streptococcal pharyngitis, rheumatic fever and rheumatic heart disease. In: Narula J et al (eds) Rheumatic fever. American Registry of Pathology, Washington, D.C., pp 41–78Google Scholar
  3. 3.
    Bisno AL (1996) Acute pharyngitis: etiology and diagnosis. Pediatrics 97:S949–S954Google Scholar
  4. 4.
    Shulman ST et al (2000) Streptococcal infections. In: Stevens D, Kaplan E (eds) Clinical aspects, microbiology, and molecular pathogenesis. Oxford University Press, New York, pp 76–101Google Scholar
  5. 5.
    World Health Organization (2004) Rheumatic fever and rheumatic heart disease: report of a WHO expert consultation. Geneva, 20 October–1 November 2001. World Health Organization, Albany, NY, USAGoogle Scholar
  6. 6.
    Stollerman GH (1997) Rheumatic fever. Lancet 349:935–942CrossRefPubMedGoogle Scholar
  7. 7.
    Kaplan EL (1980) The group A streptococcal upper respiratory tract carrier state: an enigma. J Pediatr 97:337–345PubMedGoogle Scholar
  8. 8.
    Stollerman GH (2001) Rheumatic fever in the 21st century. Clin Infect Dis 33:806–814CrossRefPubMedGoogle Scholar
  9. 9.
    Karaaslan S, Oran B, Reisli I, Erkul I (2000) Acute rheumatic fever in Konya, Turkey. Pediatr Int 42:71–75CrossRefPubMedGoogle Scholar
  10. 10.
    Olgunturk R, Aydin GB, Tunaoglu FS, Akalin N (1999) Rheumatic heart disease prevalence among schoolchildren in Ankara, Turkey. Turk J Pediatr 41:201–206PubMedGoogle Scholar
  11. 11.
    Guilherme L, Kalil J (2002) Rheumatic fever: the T cell response leading to autoimmune aggression in the heart. Autoimmun Rev 1:261–266Google Scholar
  12. 12.
    Brewerton DA, Joseph J (1976) Bunim memorial lecture. HLA-B27 and the inheritance of susceptibility to rheumatic disease. Arthritis Rheum 19:656–668PubMedGoogle Scholar
  13. 13.
    Ebringer A, Wilson C (2000) HLA molecules, bacteria and autoimmunity. J Med Microbiol 49:305–311Google Scholar
  14. 14.
    Senitzer D, Freimer EH (1984) Autoimmune mechanisms in the pathogenesis of rheumatic fever. Rev Infect Dis 6:832–839PubMedGoogle Scholar
  15. 15.
    Weidebach W, Goldberg AC, Chiarella JM, Guilherme L, Snitcowsky R, Pileggi F, Kalil J (1994) HLA class II antigens in rheumatic fever. Analysis of the DR locus by restriction fragment-length polymorphism and oligotyping. Hum Immunol 40:253–258CrossRefPubMedGoogle Scholar
  16. 16.
    Yoshinoya S, Pope RM (1980) Detection of immune complexes in acute rheumatic fever and their relationship to HLA-B5. J Clin Invest 65:136–145PubMedGoogle Scholar
  17. 17.
    Zwillich SH, Lipsky PE (1987) Molecular mimicry in the pathogenesis of rheumatic diseases. Rheum Dis Clin North Am 13:339–352PubMedGoogle Scholar
  18. 18.
    Chou HT, Chen CH, Tsai CH, Tsai FJ (2004) Association between transforming growth factor-beta 1 gene C-509T and T869C polymorphisms and rheumatic heart disease. Am Heart J 148:181–186CrossRefPubMedGoogle Scholar
  19. 19.
    Berdeli A, Celik HA, Ozyurek R, Aydin HH (2004) Involvement of immunoglobulin FcgammaRIIA and FcgammaRIIIB gene polymorphisms in susceptibility to rheumatic fever. Clin Biochem 37:925–929CrossRefPubMedGoogle Scholar
  20. 20.
    Malley R, Henneke P, Morse SC, Cieslewicz MJ, Lipsitch M, Thompson CM, Kurt-Jones E, Paton JC, Wessels MR, Golenbock DT (2003) Recognition of pneumolysin by toll-like receptor 4 confers resistance to pneumococcal infection. Proc Natl Acad Sci U S A 100:1966–1971CrossRefPubMedGoogle Scholar
  21. 21.
    Knapp S, Wieland CW, Van Veer C, Takeuchi O, Akira S, Florquin S, van der Poll T (2004) Toll-like receptor 2 plays a role in the early inflammatory response to murine pneumococcal pneumonia but does not contribute to antibacterial defense. J Immunol 172:3132–3138PubMedGoogle Scholar
  22. 22.
    O’Neill LA (2004) TLRs: Professor Mechnikov, sit on your hat. Trends Immunol 25:687–693Google Scholar
  23. 23.
    Arbour NC, Lorenz E, Schutte BC, Zabner J, Kline JN, Jones M, Frees K, Watt JL, Schwartz DA (2000) TLR4 mutations are associated with endotoxin hyporesponsiveness in humans. Nat Genet 25:187–191CrossRefPubMedGoogle Scholar
  24. 24.
    Kiechl S, Lorenz E, Reindl M, Wiedermann CJ, Oberhollenzer F, Bonora E, Willeit J, Schwartz DA (2002) Toll-like receptor 4 polymorphisms and atherogenesis. N Engl J Med 347:185–192CrossRefPubMedGoogle Scholar
  25. 25.
    Fitzgerald KA, Palsson-McDermott EM, Bowie AG, Jefferies CA, Mansell AS, Brady G, Brint E, Dunne A, Gray P, Harte MT, McMurray D, Smith DE, Sims JE, Bird TA, O’Neill LA (2001) Mal (MyD88-adapter-like) is required for toll-like receptor 4 signal transduction. Nature 413:78–83CrossRefPubMedGoogle Scholar
  26. 26.
    Horng T, Barton GM, Medzhitov R (2001) TIRAP: an adapter molecule in the toll signaling pathway. Nat Immunol 2:835–841Google Scholar
  27. 27.
    Schroder NW, Schumann RR (2005) Single nucleotide polymorphisms of toll-like receptors and susceptibility to infectious disease. Lancet Infect Dis 5:156–164PubMedGoogle Scholar
  28. 28.
    Lorenz E, Mira JP, Cornish KL, Arbour NC, Schwartz DA (2000) A novel polymorphism in the toll-like receptor 2 gene and its potential association with staphylococcal infection. Infect Immun 68:6398–6401CrossRefPubMedGoogle Scholar
  29. 29.
    Hawn TR, Verbon A, Lettinga KD, Zhao LP, Li SS, Laws RJ, Skerrett SJ, Beutler B, Schroeder L, Nachman A, Ozinsky A, Smith KD, Aderem A (2003) A common dominant TLR5 stop codon polymorphism abolishes flagellin signaling and is associated with susceptibility to Legionnaires’ disease. J Exp Med 198:1563–1572CrossRefPubMedGoogle Scholar
  30. 30.
    Medvedev AE, Lentschat A, Kuhns DB, Blanco JC, Salkowski C, Zhang S, Arditi M, Gallin JI, Vogel SN (2003) Distinct mutations in IRAK-4 confer hyporesponsiveness to lipopolysaccharide and interleukin-1 in a patient with recurrent bacterial infections. J Exp Med 198:521–531CrossRefPubMedGoogle Scholar
  31. 31.
    Picard C, Puel A, Bonnet M, Ku CL, Bustamante J, Yang K, Soudais C, Dupuis S, Feinberg J, Fieschi C, Elbim C, Hitchcock R, Lammas D, Davies G, Al-Ghonaium A, Al-Rayes H, Al-Jumaah S, Al-Hajjar S, Al-Mohsen IZ, Frayha HH, Rucker R, Hawn TR, Aderem A, Tufenkeji H, Haraguchi S, Day NK, Good RA, Gougerot-Pocidalo MA, Ozinsky A, Casanova JL (2003) Pyogenic bacterial infections in humans with IRAK-4 deficiency. Science 299:2076–2079CrossRefPubMedGoogle Scholar
  32. 32.
    Committee on Rheumatic Fever and Bacterial Endocarditis of the American Heart Association (1982) Jones criteria (revised) for guidance in the diagnosis of rheumatic fever. American Heart Association, Dallas, TX, USAGoogle Scholar
  33. 33.
    Schröder NW, Hermann C, Hamann L, Göbel UB, Hartung T, Schumann RR (2003) High frequency of polymorphism Arg753Gln of the toll-like receptor 2 gene detected by a novel allele-specific PCR. J Mol Med 81:368–372PubMedGoogle Scholar
  34. 34.
    World Health Organization (1988) Rheumatic fever and rheumatic heart disease. Report of a WHO Study Group. World Health Organization, Geneva (Technical Report Series No. 764)Google Scholar
  35. 35.
    Taranta A, Markowitz M (1989) Rheumatic fever. Kluwer, Boston, pp 19–25Google Scholar
  36. 36.
    Stevens D, Kaplan E (2000) Streptococcal infections. Clinical aspects, microbiology and molecular pathogenesis. Oxford University Press, New York, pp 102–132Google Scholar
  37. 37.
    Simpson WA, Courtney HS, Ofek I (1987) Interactions of fibronectin with streptococci: the role of fibronectin as a receptor for Streptococcus pyogenes. Rev Infect Dis 9:S351–S359Google Scholar
  38. 38.
    Kotb M, Watanabe-Ohnishi R, Wang B (1993) Analysis of the TCR V beta specificities of bacterial superantigens using PCR. ImmunoMethods 2:33–40CrossRefGoogle Scholar
  39. 39.
    Underhill DM (2004) Toll-like receptors and microbes take aim at each other. Curr Opin Immunol 16:483–487CrossRefPubMedGoogle Scholar
  40. 40.
    Schwandner R, Dziarski R, Wesche H, Rothe M, Kirschning CJ (1999) Peptidoglycan and lipoteichoic acid-induced cell activation is mediated by toll-like receptor 2. J Biol Chem 274:17406–17409CrossRefPubMedGoogle Scholar
  41. 41.
    Schröder NW, Opitz B, Lamping N, Michelsen KS, Zähringer U, Göbel UB, Schumann RR (2000) Involvement of lipopolysaccharide binding protein, CD14, and toll-like receptors in the initiation of innate immune responses by Treponema glycolipids. J Immunol 165:2683–2693PubMedGoogle Scholar
  42. 42.
    Hirschfeld M, Kirschning CJ, Schwandner R, Wesche H, Weis JH, Wooten RM, Weis JJ (1999) Cutting edge: inflammatory signaling by Borrelia burgdorferi lipoprotein is mediated by toll-like receptor 2. J Immunol 163:2382–2386PubMedGoogle Scholar
  43. 43.
    Takeuchi O, Kaufmann A, Grote K, Kawai T, Hoshino K, Morr M, Mühlradt PF, Akira S (2000) Cutting edge: preferentially the R-stereoisomer of the mycoplasmal lipopeptide macrophage-activating lipopeptide-2 activates immune cells through a toll-like receptor 2 and MYD88-dependent signaling pathway. J Immunol 164:554–557PubMedGoogle Scholar
  44. 44.
    Takeuchi O, Akira S (2002) Genetic approaches to the study of toll-like receptor function. Microbes Infect 4:887–895CrossRefPubMedGoogle Scholar
  45. 45.
    Kang TJ, Chae GT (2001) Detection of toll-like receptor 2 (TLR2) mutation in the lepromatous leprosy patients. FEMS Immunol Med Microbiol 31:53–58CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Afig Berdeli
    • 1
  • Handan Ak Celik
    • 2
  • Ruhi Özyürek
    • 3
  • Buket Dogrusoz
    • 3
  • Hikmet Hakan Aydin
    • 2
  1. 1.Laboratory of Molecular Medicine, Department of PediatricsEge University School of MedicineBornovaTurkey
  2. 2.Department of BiochemistryEge University School of MedicineBornovaTurkey
  3. 3.Department of Pediatric CardiologyEge University School of MedicineBornovaTurkey

Personalised recommendations