Cd14, Gbp1, and Pla2g2a: three major candidate genes for experimental IBD identified by combining QTL and microarray analyses.
Induction of inflammatory bowel (IBD)-like disease in mice by a targeted mutation in the Il10 gene (Il10(-/-)) is inbred strain dependent. C3H/HeJBir (C3) mice are colitis susceptible, whereas C57BL/6J (B6) mice are resistant. Genetic dissection of this susceptibility revealed 10 colitogenic quantitative trait loci (QTL). The aim of this study was to identify valuable candidate genes by a combination of QTL mapping and microarray analyses. Sixteen genes were differentially expressed between B6- and C3-Il10(-/-) mice and were located within the QTL intervals. Three major candidate genes (Cd14, Gbp1, Pla2g2a) showed prominent expression differences between B6- and C3-Il10(-/-) as well as between B6 and C3 wild-type mice, which was confirmed by semiquantitative or real-time RT-PCR. Because strain differences are known for Gbp1 and Pla2g2a, further analyses focused on Cd14. Western blot analysis revealed strain differences also on the protein level. Cd14 expression in animals with defective and intact Toll-like receptor (TLR)4 signaling (C3, C3H/HeN, B6, B6-Tlr4(tm1Aki)) make the TLR4 defect of C3 mice unlikely to be the reason for higher Cd14 expression. Less Cd14 expression in germ-free mice indicates a contribution of the microflora on Cd14 expression. Stimulation of naive peritoneal macrophages with bacterial antigens showed lower CD14 surface expression in B6 than in C3 mice. In conclusion, the large number of candidate genes was reduced to three major candidates that play an important role in inflammatory processes and immune response. Strain differences for them are already known or are shown in this study.
de Buhr
Maike F
MF
Central Animal Facility, Institute for Laboratory Animal Science, Hannover Medical School, Hannover, Germany.
Mähler
Michael
M
Geffers
Robert
R
Hansen
Wiebke
W
Westendorf
Astrid M
AM
Lauber
Jörg
J
Buer
Jan
J
Schlegelberger
Brigitte
B
Hedrich
Hans J
HJ
Bleich
Andre
A
D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology; D002478:Cells, Cultured; D003093:Colitis, Ulcerative / Q000235:genetics / Q000378:metabolism; D003106:Colon / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D054501:Group II Phospholipases A2; D015212:Inflammatory Bowel Diseases / Q000235:genetics* / Q000378:metabolism; D016753:Interleukin-10; D018950:Lipopolysaccharide Receptors / Q000235:genetics* / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D020411:Oligonucleotide Array Sequence Analysis; D010741:Phospholipases A / Q000235:genetics* / Q000378:metabolism; D040641:Quantitative Trait Loci; D012333:RNA, Messenger / Q000378:metabolism; D015203:Reproducibility of Results; D013045:Species Specificity
D000942:Antigens, Bacterial; C490288:Gbp2b protein, mouse; D018950:Lipopolysaccharide Receptors; D012333:RNA, Messenger; D016753:Interleukin-10; D010741:Phospholipases A; D054501:Group II Phospholipases A2; C516288:Pla2g2a protein, mouse; D019204:GTP-Binding Proteins