Abstract
MD-2 is an accessory protein of the Toll-like receptor (TLR)-4, necessary for assembling a receptor complex to sense low quantities of lipopolysaccharide in order to subsequently trigger innate immune responses. MD-2 and TLR-4 are expressed on a variety of immunocompetent cells. Mutations within the TLR-4 gene have been shown to attenuate immune responses against lipopolysaccharide in mice. In humans, a TLR-4 polymorphism has been associated with a higher risk for developing severe Gram-negative sepsis and with a lower risk for atherosclerosis. Since MD-2 is an essential part of the lipopolysaccharide receptor complex, we screened 20 patients that underwent surgical cancer therapy for novel MD-2 mutations by a single-strand conformation polymorphism technique. In one patient we found an A → G substitution at position 103, resulting in an amino-acid exchange from Thr 35 to Ala. Reporter gene assays revealed that this mutation resulted in a reduced lipopolysaccharide-induced signaling. The patient displayed an uneventful postoperative course, with the exception of slightly decreased TNF-α levels after in vitro stimulation with LPS as compared to wt patients. Genotyping of a further 41 patients by a newly developed Lightcycler/FRET method failed to detect any additional polymorphism carriers, indicating that this is a rare mutation.
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Acknowledgements
This work was supported by the Bundesministerium für Bildung und Forschung (BMBF, CAPNetz, Project C5) and the Deutsche Forschungsgemeinschaft (DFG) to RRS (Schr 726/1-1). We would like to thank Olfert Landt (TIB MOLBIOL, Berlin, Germany) for designing primers and fluorescent probes for Lightcycler/FRET genotyping.
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Hamann, L., Kumpf, O., Müller, M. et al. A coding mutation within the first exon of the human MD-2 gene results in decreased lipopolysaccharide-induced signaling. Genes Immun 5, 283–288 (2004). https://doi.org/10.1038/sj.gene.6364068
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DOI: https://doi.org/10.1038/sj.gene.6364068
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