Molecular Biology Reports

, Volume 38, Issue 1, pp 379–385 | Cite as

Association of polymorphisms in the human IL-10 and IL-18 genes with rheumatoid arthritis

  • Binwu Ying
  • Yunying Shi
  • Xiaofu Pan
  • Xingbo Song
  • Zhunchun Huang
  • Qian Niu
  • Bei Cai
  • Lanlan Wang


The decrease of anti-inflammatory cytokine and increase of pro-inflammatory cytokine was observed in rheumatoid arthritis (RA). Interleukin-10 (IL-10), a potent anti-inflammatory cytokine, has been demonstrated to suppress joint swelling and deformation in RA animal model. Interleukin-18 (IL-18), a widely distributed pro-inflammatory cytokine, induces the production of IFN-γ, activate NK cells, and promote inflammation. Recent studies demonstrated that the serum IL-10 and IL-18 levels may be influenced by genetics and related to susceptibility to several autoimmune diseases. In the present study, using polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) and DNA sequencing techniques, we analyzed the genotype and allele distributions of two single nucleotide polymorphisms (SNP) loci in the promoter region of IL-10 and IL-18 genes (IL-10-592 A/C and IL-18-607 A/C loci, respectively). Our results indicated that IL-10-592 allelic and genotypic frequencies were significantly different between the RA patients and normal subjects (P < 0.05). In addition, significant differences of IL-10-592 allelic and genotypic frequencies were also detected between the patients with or without anti-cyclic citrullinated peptide antibody (anti-CCP) (P < 0.05). In contrast, allelic and genotypic frequencies of IL-18-607 did not show significant difference between RA patients and normal subjects (P > 0.05) or between anti-CCP-positive and anti-CCP-negative RA patients (P > 0.05). Furthermore, ELISA detection of IL-10 and IL-18 serum levels revealed that the genotype of IL-10-592 was associated with IL-10 serum level (P < 0.05), but the genotype and allele frequency of IL-18-607 was not associated with IL-18 serum level (P > 0.05). Taken together, our findings provide new insight for the polymorphism of IL-10 gene in the pathogenesis of RA.


IL-10 IL-18 Single nucleotide polymorphisms Rheumatoid arthritis 







Rheumatoid arthritis


Single nucleotide polymorphisms


Anti-cyclic citrullinated peptide antibody


Enzyme-linked immunosorbent assay


T-helper 1


T-helper 2

Supplementary material

11033_2010_119_MOESM1_ESM.doc (28 kb)
Supplementary material 1 (DOC 27 kb)
11033_2010_119_MOESM2_ESM.ppt (936 kb)
Supplementary Fig. 1Reverse sequencing results of PCR products of IL-10-592 and IL-18-607 SNP loci. Representative results for genotypes AA (a), CC (b) and AC (c) for IL-10-592 (A) and IL-18-607 (B) SNP loci, respectively. Arrow indicates the SNP position in PCR products of IL-10 and IL-18 genes (PPT 935 kb)


  1. 1.
    Dimitrova PA, Skapenko ML, Herrmann R et al (2002) Restriction of de novo pyrimidine biosynthesis inhibits Th1 cell activation and promotes Th2 cell differentiation. J Immunol 169:3392–3399PubMedGoogle Scholar
  2. 2.
    Simon AK, Seipelt E, Wu P et al (1994) Divergent T-cell cytokine patterns in inflammatory arthritis. Proc Natl Acad Sci USA 18:8562–8566CrossRefGoogle Scholar
  3. 3.
    Cai LQ, Wang ZX, Lu WS et al (2010) A single-nucleotide polymorphism of the TNFAIP3 gene is associated with systemic lupus erythematosus in Chinese Han population. Mol Biol Rep 1:389–394CrossRefGoogle Scholar
  4. 4.
    Ji JD, Lee WJ, Kong KA et al (2010) Association of STAT4 polymorphism with rheumatoid arthritis and systemic lupus erythematosus: a meta-analysis. Mol Biol Rep 1:141–147CrossRefGoogle Scholar
  5. 5.
    Kanil KS, Hagiwara E, Yarboro CH et al (1998) Distinct patterns of cytokine secretion characterize new onset synovitis versus chronic rheumatoid arthritis. J Rheumatol 1:16–22Google Scholar
  6. 6.
    Whalen JD, Lechman EL, Carlos CA et al (1999) Adenoviral transfer of the viral IL-10 gene periarticularly to mouse paws suppresses development of collagen-induced arthritis in both injected and uninjected paws. J Immunol 162:3625–3632PubMedGoogle Scholar
  7. 7.
    Liew FY, McInnes IB (2002) Role of interleukin 18 in inflammatory response. Ann Rheum Dis 61:100–102CrossRefGoogle Scholar
  8. 8.
    Plenge RM, Seielstad M, Padyukov L et al (2007) TRAF1-C5 as a risk locus for rheumatoid arthritis—a genome wide study. N Engl J Med 12:1199–1209CrossRefGoogle Scholar
  9. 9.
    Westendorp RG, Langermans JA, Huizinga TW et al (1997) Genetic influence on cytokine production and fatal meningococcal disease. Lancet 9046:170–173CrossRefGoogle Scholar
  10. 10.
    Bai J, Jiang L, Lin M et al (2009) Association of polymorphisms in the tumor necrosis factor-alpha and interleukin-10 genes with oral lichen planus: a study in a Chinese cohort with Han ethnicity. J Interferon Cytokine Res 7:381–388CrossRefGoogle Scholar
  11. 11.
    Hikami K, Ehara Y, Hasegawa M et al (2008) Association of IL-10 receptor 2 (IL10RB) SNP with systemic sclerosis. Biochem Biophys Res Commun 3:403–407CrossRefGoogle Scholar
  12. 12.
    Tamura K, Fukuda Y, Sashio H et al (2002) IL-18 polymorphism is associated with an increased risk of Crohn’s disease. J Gastroenterol 14:111–116Google Scholar
  13. 13.
    Samsami Dehaghani A, Shahriary K et al (2009) Interleukin-18 gene promoter and serum level in women with ovarian cancer. Mol Biol Rep 8:2393–2397CrossRefGoogle Scholar
  14. 14.
    Arnett FC, Edworthy SM, Bloch DA et al (1988) The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum 31:315–324CrossRefPubMedGoogle Scholar
  15. 15.
    Jorgensen KT, Wiik A, Pedersen M et al (2008) Cytokines, autoantibodies and viral antibodies in premorbid and postdiagnostic sera from patients with rheumatoid arthritis: case control study nested in a cohort of Norwegian blood donors. Ann Rheum Dis 6:860–866CrossRefGoogle Scholar
  16. 16.
    Kawashima M, Missec P (2005) mRNA quantification of T-bet, GATA-3, IFN-gamma, and IL-4 shows a defective Th1 immune response in the peripheral blood from rheumatoid arthritis patients: link with disease activity. J Clin Immunol 3:209–214CrossRefGoogle Scholar
  17. 17.
    Nistala K, Wedderburn LR (2009) Th17 and regulatory T cells: rebalancing pro- and anti-inflammatory forces in autoimmune arthritis. Rheumatology 6:602–606CrossRefGoogle Scholar
  18. 18.
    Charbonnier LM, Han WGH, Quentin J et al. (2009) Adoptive transfer of IL-10-secreting CD4+CD49b+ regulatory T cells suppresses ongoing arthritis. J Autoimmun. doi:10.1016/j.jaut.2009.10.003
  19. 19.
    Panoulas VF, Smith JP, Stavropoulos-Kalinoglou A et al (2009) Lack of an association of GNB3 C825T polymorphism and blood pressure in patients with rheumatoid arthritis. Clin Exp Hypertens 5:428–439CrossRefGoogle Scholar
  20. 20.
    Assmann G, Voswinkel J, Mueller M et al (2009) Association of rheumatoid arthritis with Mdm2 SNP309 and genetic evidence for an allele-specific interaction between MDM2 and p53 P72R variants: a case control study. Clin Exp Rheumatol 4:615–619Google Scholar
  21. 21.
    Burkhardt J, Petit-Teixeira E, Teixeira VH et al (2009) Association of the X-chromosomal genes TIMP1 and IL9R with rheumatoid arthritis. J Rheumatol 10:2149–2157CrossRefGoogle Scholar
  22. 22.
    Suarez-Gestal M, Calaza M, Dieguez-Gonzalez R et al (2009) Rheumatoid arthritis does not share most of the newly identified systemic lupus erythematosus genetic factors. Arthritis Rheum 9:2558–2564CrossRefGoogle Scholar
  23. 23.
    Dieguez-Gonzalez R, Akar S, Calaza M et al (2009) Lack of association with rheumatoid arthritis of selected polymorphisms in 4 candidate genes: CFH, CD209, eotaxin-3, and MHC2TA. J Rheumatol 8:1590–1595CrossRefGoogle Scholar
  24. 24.
    Kirsten H, Petit-Teixeira E, Scholz M et al (2009) Association of MICA with rheumatoid arthritis independent of known HLA-DRB1 risk alleles in a family-based and a case control study. Arthritis Res Ther 3:R60CrossRefGoogle Scholar
  25. 25.
    Fan LY, Zong M, Lu TB et al (2009) Association of the PADI4 gene polymorphism and HLA-DRB1 shared epitope alleles with rheumatoid arthritis. Zhonghua Yi Xue Yi Chuan Xue Za Zhi 1:57–61Google Scholar
  26. 26.
    Moore KW, Malefyt R, Coffinan RL et al (2001) Interleukin-10 and the interleukin-10 receptor. Annu Rev Immunol 19:683–765CrossRefPubMedGoogle Scholar
  27. 27.
    Turner DM, Williams DM, Sankaran D et al (1997) An investigation of polymorphism in the interleukin-10 gene promoter. Eur J Immunogenet 1:1–8Google Scholar
  28. 28.
    Rees LE, Wood NA, Gillespie KM et al (2002) The interleukin-10–1082 G/A polymorphism: allele frequency in different populations and functional significance. Cell Mol Life Sci 3:559–560Google Scholar
  29. 29.
    Tarzi M, Klunker S, Texier C et al (2006) Induction of interleukin-10 and suppressor of cytokine signalling-3 gene expression following peptide immunotherapy. Clin Exp Allergy 4:465–474CrossRefGoogle Scholar
  30. 30.
    Li XD, Dong TH, Ni H (2006) Research progress of relation between single nucleotide polymorphisms in IL-10 Gene and disease. Int Genet J 5:368–371Google Scholar
  31. 31.
    Crawley E, Kay R, Siliborne J et al (1999) Polymorphic haplotypes of the IL-10 5’ flanking region determine valuable IL-10 transcription and are associated with particular phenotypes of juvenile rheumatoid arthritis. Arthritis Rheum 42:1101–1108CrossRefPubMedGoogle Scholar
  32. 32.
    Dinarello CA, Fantuzzi G (2003) Interleukin-18 and host defense against infection. J Infect Dis 187:370–384CrossRefGoogle Scholar
  33. 33.
    Favilli F, Anzilotti C, Martinelli L et al (2009) IL-18 activity in systemic lupus erythematosus. Ann N Y Acad Sci 1173:301–309CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Binwu Ying
    • 1
  • Yunying Shi
    • 2
  • Xiaofu Pan
    • 1
  • Xingbo Song
    • 1
  • Zhunchun Huang
    • 1
  • Qian Niu
    • 1
  • Bei Cai
    • 1
  • Lanlan Wang
    • 1
  1. 1.Department of Laboratory Medicine, West China HospitalSichuan UniversityChengduPeople’s Republic of China
  2. 2.Department of Nephrology, West China HospitalSichuan UniversityChengduPeople’s Republic of China

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