Linkage of Crohn's disease-related serological phenotypes: NFKB1 haplotypes are associated with anti-CBir1 and ASCA, and show reduced NF-kappaB activation

Gut. 2009 Jan;58(1):60-7. doi: 10.1136/gut.2008.156422. Epub 2008 Oct 2.


Background and aims: Genetics studies of the serum expression of antibodies to microbial antigens may yield important clues to the pathogenesis of Crohn's disease. Our aim was to conduct a linkage study using expression of anti-CBir1, anti-I2, anti-OmpC and ASCA as quantitative traits.

Methods: Expression of antibodies to microbial antigens was measured by enzyme-linked immunosorbant assay (ELISA) and a standard approximately 10 cM whole genome microsatellite study was conducted. Single nucleotide polymorphism genotyping was performed using either Illumina or TaqMan MGB technology. Nuclear factor Kappa B (NF-kappaB) activation in cells from Epstein-Barr virus (EBV)-transformed cell lines was assessed using an electrophoretic mobility shift assay and protein was measured using ELISA and western blotting.

Results: Evidence for linkage to anti-CBir1 expression was detected on human chromosome 4 (logarithm of odds (LOD) 1.82 at 91 cM). We therefore directly proceeded to test the association of haplotypes in NFKB1, a candidate gene. One haplotype, H1, was associated with anti-CBir1 (p = 0.003) and another, H3, was associated with ASCA (p = 0.023). Using cell lines from Crohn's disease patients with either H1 or H3, NF-kappaB activation and NF-kappaB p105 and p50 production were significantly lower for patients with H1 compared to patients with H3.

Conclusions: These results suggest that NFKB1 haplotypes induce dysregulation of innate immune responses by altering NF-kappaB expression. The results also show the use of EBV-transformed lymphoblastoid cell lines to conduct phenotypic studies of genetic variation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Bacterial / blood*
  • Antigens, Bacterial / immunology
  • Case-Control Studies
  • Cell Line, Transformed
  • Cell Transformation, Viral
  • Chromosomes, Human, Pair 4 / genetics
  • Crohn Disease / genetics*
  • Crohn Disease / immunology
  • Down-Regulation
  • Flagellin / immunology
  • Genetic Linkage
  • Haplotypes
  • Humans
  • NF-kappa B / metabolism*
  • NF-kappa B p50 Subunit / genetics*
  • Phenotype
  • Polymorphism, Single Nucleotide
  • Saccharomyces cerevisiae / immunology


  • Antibodies, Bacterial
  • Antigens, Bacterial
  • CBir1 flagellin
  • NF-kappa B
  • NF-kappa B p50 Subunit
  • NFKB1 protein, human
  • Flagellin