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TLR4 mutations are associated with endotoxin hyporesponsiveness in humans

Abstract

There is much variability between individuals in the response to inhaled toxins, but it is not known why certain people develop disease when challenged with environmental agents and others remain healthy. To address this, we investigated whether TLR4 (encoding the toll-like receptor-4), which has been shown to affect lipopolysaccharide (LPS) responsiveness in mice1,2, underlies the variability in airway responsiveness to inhaled LPS in humans3. Here we show that common, co-segregating missense mutations (Asp299Gly and Thr399Ile) affecting the extracellular domain of the TLR4 receptor are associated with a blunted response to inhaled LPS in humans. Transfection of THP-1 cells demonstrates that the Asp299Gly mutation (but not the Thr399Ile mutation) interrupts TLR4-mediated LPS signalling. Moreover, the wild-type allele of TLR4 rescues the LPS hyporesponsive phenotype in either primary airway epithelial cells or alveolar macrophages obtained from individuals with the TLR4 mutations. Our findings provide the first genetic evidence that common mutations in TLR4 are associated with differences in LPS responsiveness in humans, and demonstrate that gene-sequence changes can alter the ability of the host to respond to environmental stress.

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Figure 1: A common missense mutation in human TLR4.
Figure 2: Airway responsiveness to inhaled LPS and TLR4 genotype.
Figure 3: Functional significance of TLR4 mutations in THP-1 cells.
Figure 4: Functional significance of TLR4 mutations in primary human epithelial cells.
Figure 5: Rescue of the LPS hyporesponsive phenotype.

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References

  1. Poltorak, A. et al. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 282, 2085–2088 (1998).

    CAS  Article  Google Scholar 

  2. Hoshino, K. et al. Cutting edge: toll-like receptor 4 (TLR4)-deficient mice are hyporesponsive to lipopolysacchaide: evidence for TLR4 as the Lps gene product. J. Immunol. 162, 3749–3752 (1999).

    CAS  Google Scholar 

  3. Kline, J.N. et al. Variable airway responsiveness to inhaled lipopolysaccharide. Am. J. Respir. Crit. Care Med. 160, 297–303 (1999).

    CAS  Article  Google Scholar 

  4. Michel, O. et al. Severity of asthma is related to endotoxin in house dust. Am. J. Respir. Crit. Care Med. 154, 1641–1646 (1996).

    CAS  Article  Google Scholar 

  5. Schwartz, D.A. et al. The role of endotoxin in grain dust-induced lung disease. Am. J. Respir. Crit. Care Med. 152, 603–608 (1995).

    CAS  Article  Google Scholar 

  6. Schwartz, D.A. et al. Determinants of longitudinal changes in spirometric functions among swine confinement operators and farmers. Am. J. Respir. Crit. Care Med. 151, 47–53 (1995).

    CAS  Article  Google Scholar 

  7. Bonner, J.C. et al. Induction of the lung myofibroblast PDGF receptor system by urban ambient particles from Mexico City. Am. J. Respir. Cell Mol. Biol. 19, 672–680 (1998).

    CAS  Article  Google Scholar 

  8. Santamaria, P., Gehrz, R.C., Bryan, M.K. & Barbosa, J.J. Involvement of class II MHC molecules in the LPS-induction of IL-1/TNF secretions by human monocytes. Quantitative differences at the polymorphic level. J. Immunol. 143, 913–922 (1989).

    CAS  PubMed  Google Scholar 

  9. Kuhns, D.B., Long Priel, D.A. & Gallin, J.I. Endotoxin and IL-1 hyporesponsiveness in a patient with recurrent bacterial infections. J. Immunol. 158, 3959–3964 (1997).

    CAS  Google Scholar 

  10. Chow, J.C., Young, D.W., Golenbock, D.T., Christ, W.J. & Gusovsky, F. Toll-like receptor-4 mediates lipopolysaccharide-induced signal transduction. J. Biol. Chem. 274, 10689–10692 (1999).

    CAS  Article  Google Scholar 

  11. Shimazu, R. et al. MD-2, a molecule that confers lipopolysaccharide responsiveness on toll-like receptor 4. J. Exp. Med. 189, 1777–1782 (1999).

    CAS  Article  Google Scholar 

  12. Medzhitov, R., Preston-Hurlburt, P. & Janeway, C.A. Jr A human homologue of the Drosophila Toll protein signals activation of adaptive immunity. Nature 388, 394–397 (1997).

    CAS  Article  Google Scholar 

  13. Lidral, A.C. et al. Association of MSX1 and TGFB3 with nonsyndromic clefting in humans. Am. J. Hum. Genet. 63, 557–568 (1998).

    CAS  Article  Google Scholar 

  14. NIH-CEPH (Collaborative Mapping Group) A comprehensive genetic linkage map of the human genome. Science 258, 67–68 (1992).

  15. Becker, S. et al. Compartmentalization of the inflammatory response to inhaled grain dust. Am. J. Respir. Crit. Care Med. 160, 1309–1318 (1999).

    CAS  Article  Google Scholar 

  16. Brigham, K.L. & Meyrick, B. State of the art: endotoxin and lung injury. Am. Rev. Respir. Dis. 133, 913–927 (1986).

    CAS  PubMed  Google Scholar 

  17. OBrien, A.D., Rosenstreich, D.L. & Taylor, B.A. Control of natural resistance to Salmonella typhimurium and Leishmania donovani in mice by closely linked but distinct genetic loci. Nature 287, 440–442 (1980).

    CAS  Article  Google Scholar 

  18. Gibrat, J.F., Garnier, J. & Robson, B. Further developments of protein secondary structure prediction using information theory. New parameters and consideration of residue pairs. J. Mol. Biol. 198, 425–443 (1987).

    CAS  Article  Google Scholar 

  19. Rask-Andersen, A., Malmberg, P. & Lundholm, M. Endotoxin levels in farming: absence of symptoms despite high exposure level. Br. J. Ind. Med. 46, 412–416 (1989).

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Warren, W. et al. Detection of mutations by single-strand conformation polymorphism (SSCP) analysis and SSCP-hybrid methods. in Current Protocols in Human Genetics (eds Dracopoli, N.C. et al.) 7.41–7.4.23 (John Wiley & Sons, New York, 1997).

    Chapter  Google Scholar 

  21. Zabner, J., Zeiher, B.G., Friedman, E. & Welsh, M.J. Adenovirus-mediated gene transfer to ciliated airway epithelia requires prolonged incubation time. J. Virol. 70, 6694–7003 (1996).

    Google Scholar 

  22. Rock, F.L., Hardiman, G., Timans, J.C., Kastelein, R.A. & Bazan, J.F. A family of human receptors structurally related to Drosophila Toll. Proc. Natl Acad. Sci. USA 95, 588–593 (1998).

    CAS  Article  Google Scholar 

  23. Davidson, B.L. et al. Expression of Escherichia coli β-galactosidease and rat HPRT in the CNS of Macaca mulatta following adenoviral mediated gene transfer. Exp. Neurol. 125, 258–267 (1994).

    CAS  Article  Google Scholar 

  24. Fasbender, A. et al. Incorporation of adenovirus in calcium phosphate precipitates enhances gene transfer to airway epithelia in vitro and in vivo. J. Clin. Invest. 102, 184–193 (1998).

    CAS  Article  Google Scholar 

  25. Fisher, L.D. & van Belle, G. Biostatistics—A Methodology for the Health Sciences (Wiley, New York, 1993).

    Google Scholar 

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Acknowledgements

We thank C. Galer, T. Grunst, M. Malik, N. Meyer, M. Monick, K. Schneider and S. Swartz for technical assistance; the University of Iowa Airway Epithelia Cell Culture Core and Gene Transfer Vector Core; and M. Welsh and J. Murray for discussions. This study was supported by grants from the Department of Veterans' Affairs (Merit Review), the National Institute of Environmental Health Sciences (ES06537, ES07498 and ES09607), the National Heart Lung and Blood Institute (HL62628 and HL64855) and the General Clinical Research Centers Program (RR00059).

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Correspondence to David A. Schwartz.

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Arbour, N., Lorenz, E., Schutte, B. et al. TLR4 mutations are associated with endotoxin hyporesponsiveness in humans. Nat Genet 25, 187–191 (2000). https://doi.org/10.1038/76048

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