Linkage of Crohn's disease-related serological phenotypes: NFKB1 haplotypes are associated with anti-CBir1 and ASCA, and show reduced NF-kappaB activation.
OBJECTIVE
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.
Takedatsu
H
H
Division of Gastroenterology, Inflammatory Bowel Disease Center, Immunobiology Research Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA. targans@cshs.org
Taylor
K D
KD
Mei
L
L
McGovern
D P B
DP
Landers
C J
CJ
Gonsky
R
R
Cong
Y
Y
Vasiliauskas
E A
EA
Ippoliti
A
A
Elson
C O
CO
Rotter
J I
JI
Targan
S R
SR
D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000276:immunology; D016022:Case-Control Studies; D002461:Cell Line, Transformed; D002472:Cell Transformation, Viral; D002894:Chromosomes, Human, Pair 4 / Q000235:genetics; D003424:Crohn Disease / Q000235:genetics* / Q000276:immunology; D015536:Down-Regulation; D005408:Flagellin / Q000276:immunology; D008040:Genetic Linkage; D006239:Haplotypes; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism*; D052002:NF-kappa B p50 Subunit / Q000235:genetics*; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D012441:Saccharomyces cerevisiae / Q000276:immunology
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; C502160:CBir1 flagellin; D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; C489548:NFKB1 protein, human; D005408:Flagellin