Z Gastroenterol 2005; 43 - 68
DOI: 10.1055/s-2005-869715

Toll-like receptor 4 and NOD2/CARD15 mutations in Hungarian patients with Crohn's disease: phenotype-genotype correlations

L Lakatos 1, P Lakatos 2, F Szalay 2, C Willheim-Polli 3, C Österreicher 3, Z Tulassay 4, T Molnar 5, W Reinisch 3, J Papp 2, Hungarian IBD Study Group 2, G Mozsik 6 P Ferenci 3,
  • 11st Department of Medicine, Csolnoky F. County Hospital, Veszprem, Hungary
  • 21st Department of Medicine, Semmelweis University, Budapest, Hungary
  • 3Department of Internal Medicine 4, University of Vienna, Austria
  • 41st Department of Medicine, Semmelweis University, Budapest, Hungary
  • 51st Department of Medicine, University of Szeged, Szeged, Hungary
  • 61st Department of Medicine, University of Pecs, Pecs, Hungary

Aim: Mutations of NOD2/CARD15 gene predispose for Crohn disease (CD) and are associated with fibrostenosing behaviour. Since NOD2/CARD15 is involved in the recognition of bacterial antigens including the pathway of toll like receptor 4 (TLR4) functional D299G polymorphism of TLR4 may be another genetic modifier for CD. In view of the large geographical differences in frequency of these genetic markers and absence of data in Central-European patients, common NOD2/CARD15 mutations and D299G-TLR4 polymorphism were determined in Hungarian CD patients.

Methods: DNA of 527 unrelated patients with CD (m/f: 265/262, age: 37.1 (SD 7.6) years) and of 200 healthy subjects was screened for possible NOD2/CARD15 mutations by denaturing-HPLC (confirmed by direct sequencing). TLR4-D299G was tested by PCR-RFLP. Comparisons between groups were made by Chi2 test.

Results: NOD2/CARD15 mutations were found in 185 patients (35.1%) and in 33 controls (16.5%, p<0.0001). SNP8/R702W (10.8% vs. 6%, P=0.02), SNP13/3020insC (19.4% vs. 5%, P<0.0001) and exon4 R703C (2.1% vs. 0%, p=0.02) mutations were more frequent in CD, while the frequency of SNP12/G908R was not increased. The frequency of TLR4 D299G was not different in CD and controls (9.9% vs. 12.0%). Variant NOD2/CARD15 allele was associated with an increased risk for CD (ORhet=1.71, 95%CI=1.12–2.6, P=0.0001, ORtwo-risk-alleles=25.2, 95%CI=4.37–00, p<0.0001), younger disease onset (carrier:26.4 vs. non-carrier:29.8years, P=0.0006), ileal involvement (81.9% vs. 69.5%, OR=1.99, 95%CI=1.29–3.08, P=0.02, in presence of NOD2/CARD15 and TLR4: 86.7% vs. 64.8%), stricturing behavior (OR=1.69, 95%CI=1.13–2.55, P=0.026) and increased need for resection (OR=1.71, 95% CI: 1.13–2.62, P=0.01), but not with duration, extraintestinal manifestations, familial disease or smoking. TLR4 exhibited a modifier effect: age of onset in wt/TLR4 D299G carriers: 27.4 years vs. NOD2mut/TLR D299G: 23 years (p=0.06), in NOD2mut/wt: 26.4 years.

Conclusion: These results confirm in large cohorts of Hungarian CD patients the association of variant NOD2/CARD15 (R702W, R703C and 3020 insC) alleles with younger disease onset, ileal disease, stricturing disease behavior. In contrast, although the frequency of TLR4 D299G polymorphism was not different from controls, NOD2/TLR mutation carriers tended to present at earlier age.