Defective IL-1A expression in patients with Crohn's disease is related to attenuated MAP3K4 signaling.
Crohn's disease (CD) is characterized by an aberrant immune response to bacterial products stimulating TLR, in genetically susceptible hosts. Next to mutations in the TLR signaling molecule NOD2, several other immune response- and autophagy-genes contribute to CD. Since only 10-20% of cases can be explained by a NOD2 defect, we searched for additional TLR-related disease-causing factors. We analyzed the LPS response of peripheral blood mononuclear cells from 23 CD patients in remission, compared to 16 controls in a time course experiment. Individuals with any of the three major contributing NOD2 mutations were excluded. Overall, the LPS-responsive gene transcript levels, determined by low density arrays, were significantly lower in CD patients. In particular IL-1A expression was severely reduced in CD patients (ninefold reduction, p=0.001). Quantification of several important TLR4 signal transducers and cytokines identified MAP3K4 as a candidate signaling molecule with reduced expression in CD patients, which might explain the low IL-1A expression. Silencing of MAP3K4 by lentiviral shRNA transduction indeed showed that the expression of IL-1A was specifically dependent on this kinase. Furthermore, the expression of GSK3β, an inhibitor of MAP3K4, was increased in CD patients. In conclusion, we identified a novel TLR signaling defect in CD patients involving MAP3K4 and IL-1A. This confirms the hypothesis that CD patients, despite their massive intestinal inflammation, suffer from a relative immune deficiency in TLR-mediated cytokine production.
VU University Medical Center, Dept. of Molecular Cell Biology & Immunology, Amsterdam, The Netherlands. t.vanderpouwkraan@vumc.nl
C T M
CT
van der Pouw Kraan
J M
JM
Baggen
A A
AA
van Bodegraven
C J J
CJ
Mulder
A
A
Zwiers
D
D
Geerts
G
G
Kraal
A J
AJ
Horrevoets
G
G
Bouma
D000328:Adult; D003424:Crohn Disease / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D016207:Cytokines / Q000235:genetics / Q000276:immunology; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D053582:Interleukin-1alpha / Q000235:genetics* / Q000378:metabolism; D007700:Kinetics; D008070:Lipopolysaccharides / Q000276:immunology; D048768:MAP Kinase Kinase Kinase 4 / Q000235:genetics / Q000378:metabolism*; D008297:Male; D034622:RNA Interference; D015398:Signal Transduction; D051197:Toll-Like Receptor 4 / Q000276:immunology / Q000378:metabolism
D016207:Cytokines; D053582:Interleukin-1alpha; D008070:Lipopolysaccharides; D051197:Toll-Like Receptor 4; D048768:MAP Kinase Kinase Kinase 4