Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Toll-like receptor-4 (TLR4) signaling augments chemokine-induced neutrophil migration by modulating cell surface expression of chemokine receptors

Nature Medicine volume 9, pages 315–321 (2003)Cite this article

An Erratum to this article was published on 01 April 2003

Abstract

Polymorphonuclear leukocytes (PMNs) are critical effector cells of the innate immune system that protect the host by migrating to inflammatory sites and killing pathogenic microbes. We addressed the role of chemokine receptor desensitization induced by G-protein-coupled receptor kinases (GRKs) in the feedback control of PMN migration. We show that the chemokine macrophage inflammatory protein-2 (MIP-2) induces GRK2 and GRK5 expression in PMNs through phosphoinositide-3-kinase (PI3K)-γ signaling. We also show that lipopolysaccharide (LPS)-activated signaling through the Toll-like receptor (TLR)-4 pathway transcriptionally downregulates the expression of GRK2 and GRK5 in response to MIP-2. The reduced expression of GRKs lowers chemokine receptor desensitization and markedly augments the PMN migratory response. These data indicate that TLR4 modulation of PMN surface chemokine receptor expression subsequent to the downregulation of GRK2 and GRK5 expression is a critical determinant of PMN migration.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Effect of LPS on GRK2 and GRK5 expression.
Figure 2: Effect of LPS on MIP-2–induced CXCR2 internalization.
Figure 3: Effect of LPS on MIP-2–induced PMN migration and in vivo expression of GRK2 and GRK5.
Figure 4: Effects of LPS treatment on chemokine-induced GRK2 and GRK5 expression, CXCR2 internalization, and migration of human PMNs.
Figure 5: Effect of inhibitors on LPS and chemokine regulation of GRK2 and GRK5 expression.
Figure 6: Model of TLR4 and chemokine receptor crosstalk.

Similar content being viewed by others

References

  1. Ulevitch, R.J. Toll gates for pathogen selection. Nature 401, 755–756 (1999).

    Article  CAS  Google Scholar 

  2. Baggiolini, M. Chemokines and leukocyte traffic. Nature 392, 565–568 (1998).

    Article  CAS  Google Scholar 

  3. Lefkowitz, R.J. G protein-coupled receptors. III. New roles for receptor kinases and β-arrestins in receptor signaling and desensitization. J. Biol. Chem. 273, 18677–18680 (1998).

    Article  CAS  Google Scholar 

  4. Gainetdinov, R.R. et al. Muscarinic supersensitivity and impaired receptor desensitization in G protein-coupled receptor kinase 5-deficient mice. Neuron 24, 1029–1036 (1999).

    Article  CAS  Google Scholar 

  5. Benovic, J.L., DeBlasi, A., Stone, W.C., Caron, M.G. & Lefkowitz, R.J. β-adrenergic receptor kinase: primary structure delineates a multigene family. Science 246, 235–240 (1989).

    Article  CAS  Google Scholar 

  6. Hisatomi, O. et al. A novel subtype of G-protein-coupled receptor kinase, GRK7, in teleost cone photoreceptors. FEBS Lett. 424, 159–164 (1998).

    Article  CAS  Google Scholar 

  7. Penn, R.B., Pronin, A.N. & Benovic, J.L. Regulation of G protein-coupled receptor kinases. Trends Cardiovasc. Med. 10, 81–89 (2000).

    Article  CAS  Google Scholar 

  8. Penn, R.B. & Benovic, J.L. Structure of the human gene encoding the β-adrenergic receptor kinase. J. Biol. Chem. 269, 14924–14930 (1994).

    CAS  PubMed  Google Scholar 

  9. McGraw, D.W. & Liggett, S.B. Heterogeneity in β-adrenergic receptor kinase expression in the lung accounts for cell-specific desensitization of the β2-adrenergic receptor. J. Biol. Chem. 272, 7338–7344 (1997).

    Article  CAS  Google Scholar 

  10. Eckhart, A.D., Ozaki, T., Tevaearai, H., Rockman, H.A. & Koch, W.J. Vascular-targeted overexpression of G protein-coupled receptor kinase-2 in transgenic mice attenuates β-adrenergic acceptor signaling and increases resting blood pressure. Mol. Pharmacol. 61, 749–758 (2002).

    Article  CAS  Google Scholar 

  11. Rietschel, E.T. et al. Bacterial endotoxin: molecular relationships of structure to activity and function. FASEB J. 8, 217–225 (1994).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  13. Qureshi, S.T. et al. Endotoxin-tolerant mice have mutations in Toll-like receptor 4 (Tlr4). J. Exp. Med. 189, 615–625 (1999).

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  15. Sasaki, T. et al. Function of PI3Kγ in thymocyte development, T cell activation, and neutrophil migration. Science 287, 1040–1046 (2000).

    Article  CAS  Google Scholar 

  16. Huang, J., Wang, M.D., Lenz, S., Gao, D. & Kaltenboeck, B. IL-12 administered during Chlamydia psittaci lung infection in mice confers immediate and long-term protection and reduces macrophage inflammatory protein-2 level and neutrophil infiltration in lung tissue. J. Immunol. 162, 2217–2226 (1999).

    CAS  PubMed  Google Scholar 

  17. Xing, Z. et al. IL-6 is an antiinflammatory cytokine required for controlling local or systemic acute inflammatory responses. J. Clin. Invest. 101, 311–320 (1998).

    Article  CAS  Google Scholar 

  18. Medzhitov, R. Toll-like receptors and innate immunity. Nat. Rev. Immunol. 1, 135–145 (2001).

    Article  CAS  Google Scholar 

  19. May, M.J. et al. Selective inhibition of NFκB activation by a peptide that blocks the interaction of NEMO with the IκB kinase complex. Science 289, 1550–1554 (2000).

    Article  CAS  Google Scholar 

  20. Favata, M.F. et al. Identification of a novel inhibitor of mitogen-activated protein kinase kinase. J. Biol. Chem. 273, 18623–18632 (1998).

    Article  CAS  Google Scholar 

  21. Lloyd, A.R. et al. Granulocyte-colony stimulating factor and lipopolysaccharide regulate the expression of interleukin 8 receptors on polymorphonuclear leukocytes. J. Biol. Chem. 270, 28188–28192 (1995).

    Article  CAS  Google Scholar 

  22. Khandaker, M.H. et al. CXCR1 and CXCR2 are rapidly down-modulated by bacterial endotoxin through a unique agonist-independent, tyrosine kinase-dependent mechanism. J. Immunol. 161, 1930–1938 (1998).

    CAS  PubMed  Google Scholar 

  23. Kaplan, A.P. Chemokines, chemokine receptors and allergy. Int. Arch. Allergy Immunol. 124, 423–431 (2001).

    Article  CAS  Google Scholar 

  24. Lira, S. Lessons from gene modified mice. Forum (Genova) 9, 286–298 (1999).

    CAS  Google Scholar 

  25. Cacalano, G. et al. Neutrophil and B cell expansion in mice that lack the murine IL-8 receptor homolog. Science 265, 682–684 (1994).

    Article  CAS  Google Scholar 

  26. Wiekowski, M.T. et al. Disruption of neutrophil migration in a conditional transgenic model: evidence for CXCR2 desensitization in vivo. J. Immunol. 167, 7102–7110 (2001).

    Article  CAS  Google Scholar 

  27. Khandaker, M.H. et al. Metalloproteinases are involved in lipopolysaccharide- and tumor necrosis factor-α-mediated regulation of CXCR1 and CXCR2 chemokine receptor expression. Blood 93, 2173–2185 (1999).

    CAS  PubMed  Google Scholar 

  28. Kalmar, J.R. & Van Dyke, T.E. Effect of bacterial products on neutrophil chemotaxis. Methods Enzymol. 236, 58–87 (1994).

    Article  CAS  Google Scholar 

  29. Hunter, R. Standardization of the chloramine-T method of protein iodination. Proc. Soc. Exp. Biol. Med. 133, 989–992 (1970).

    Article  CAS  Google Scholar 

  30. Yoshizumi, M., Abe, J., Haendeler, J., Huang, Q. & Berk, B.C. Src and Cas mediate JNK activation but not ERK1/2 and p38 kinases by reactive oxygen species. J. Biol. Chem. 275, 11706–11712 (2000).

    Article  CAS  Google Scholar 

  31. Bennett, B.L. et al. SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase. Proc. Natl. Acad. Sci. USA 98, 13681–13686 (2001).

    Article  CAS  Google Scholar 

  32. Szucs, S., Varga, C., Ember, I. & Kertai, P. The separation of the granulocytes from different rat strains. A comparative study. J. Immunol. Methods 167, 245–251 (1994).

    Article  CAS  Google Scholar 

  33. Fan, J., Frey, R.S., Rahman, A. & Malik, A.B. Role of neutrophil NADPH oxidase in the mechanism of tumor necrosis factor-α-induced NFκB activation and intercellular adhesion molecule-1 expression in endothelial cells. J. Biol. Chem. 277, 3404–3411 (2002).

    Article  CAS  Google Scholar 

  34. Fan, J. et al. Regulation of Toll-like receptor 4 expression in the lung following hemorrhagic shock and lipopolysaccharide. J. Immunol. 168, 5252–5259 (2002).

    Article  CAS  Google Scholar 

  35. Johnston, J.A. et al. Human T lymphocyte chemotaxis and adhesion induced by vasoactive intestinal peptide. J. Immunol. 153, 1762–1768 (1994).

    CAS  PubMed  Google Scholar 

  36. Edwards, J.C., Sedgwick, A.D. & Willoughby, D.A. The formation of a structure with the features of synovial lining by subcutaneous injection of air: an in vivo tissue culture system. J. Pathol. 134, 147–156 (1981).

    Article  CAS  Google Scholar 

  37. Fan, G.H., Yang, W., Wang, X.J., Qian, Q. & Richmond, A. Identification of a motif in the carboxyl terminus of CXCR2 that is involved in adaptin 2 binding and receptor internalization. Biochemistry 40, 791–800 (2001).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by US National Institutes of Health grants T32 HL07829, HL45638, HL60678 and HL27016.

Author information

Authors and Affiliations

  1. Department of Pharmacology, University of Illinois College of Medicine, Chicago, Illinois, USA

    Jie Fan & Asrar B. Malik

Authors
  1. Jie Fan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Asrar B. Malik
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Asrar B. Malik.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fan, J., Malik, A. Toll-like receptor-4 (TLR4) signaling augments chemokine-induced neutrophil migration by modulating cell surface expression of chemokine receptors. Nat Med 9, 315–321 (2003). https://doi.org/10.1038/nm832

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm832

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing