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. 1995 Sep 26;92(20):9288–9292. doi: 10.1073/pnas.92.20.9288

CD14 enhances cellular responses to endotoxin without imparting ligand-specific recognition.

R L Delude 1, R Savedra Jr 1, H Zhao 1, R Thieringer 1, S Yamamoto 1, M J Fenton 1, D T Golenbock 1
PMCID: PMC40970  PMID: 7568119

Abstract

Binding of the lipid A portion of bacterial lipopolysaccharide (LPS) to leukocyte CD14 activates phagocytes and initiates the septic shock syndrome. Two lipid A analogs, lipid IVA and Rhodobacter sphaeroides lipid A (RSLA), have been described as LPS-receptor antagonists when tested with human phagocytes. In contrast, lipid IVA activated murine phagocytes, whereas RSLA was an LPS antagonist. Thus, these compounds displayed a species-specific pharmacology. To determine whether the species specificity of these LPS antagonists occurred as a result of interactions with CD14, the effects of lipid IVA and RSLA were examined by using human, mouse, and hamster cell lines transfected with murine or human CD14 cDNA expression vectors. These transfectants displayed sensitivities to lipid IVA and RSLA that reflected the sensitivities of macrophages of similar genotype (species) and were independent of the source of CD14 cDNA. For example, hamster macrophages and hamster fibroblasts transfected with either mouse or human-derived CD14 cDNA responded to lipid IVA and RSLA as LPS mimetics. Similarly, lipid IVA and RSLA acted as LPS antagonists in human phagocytes and human fibrosarcoma cells transfected with either mouse or human-derived CD14 cDNA. Therefore, the target of these LPS antagonists, which is encoded in the genomes of these cells, is distinct from CD14. Although the expression of CD14 is required for macrophage-like sensitivity to LPS, CD14 cannot discriminate between the lipid A moieties of these agents. We hypothesize that the target of the LPS antagonists is a lipid A recognition protein which functions as a signaling receptor that is triggered after interaction with CD14-bound LPS.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arditi M., Zhou J., Dorio R., Rong G. W., Goyert S. M., Kim K. S. Endotoxin-mediated endothelial cell injury and activation: role of soluble CD14. Infect Immun. 1993 Aug;61(8):3149–3156. doi: 10.1128/iai.61.8.3149-3156.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Delude R. L., Fenton M. J., Savedra R., Jr, Perera P. Y., Vogel S. N., Thieringer R., Golenbock D. T. CD14-mediated translocation of nuclear factor-kappa B induced by lipopolysaccharide does not require tyrosine kinase activity. J Biol Chem. 1994 Sep 2;269(35):22253–22260. [PubMed] [Google Scholar]
  3. Dendorfer U., Oettgen P., Libermann T. A. Multiple regulatory elements in the interleukin-6 gene mediate induction by prostaglandins, cyclic AMP, and lipopolysaccharide. Mol Cell Biol. 1994 Jul;14(7):4443–4454. doi: 10.1128/mcb.14.7.4443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Frey E. A., Miller D. S., Jahr T. G., Sundan A., Bazil V., Espevik T., Finlay B. B., Wright S. D. Soluble CD14 participates in the response of cells to lipopolysaccharide. J Exp Med. 1992 Dec 1;176(6):1665–1671. doi: 10.1084/jem.176.6.1665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Golenbock D. T., Hampton R. Y., Qureshi N., Takayama K., Raetz C. R. Lipid A-like molecules that antagonize the effects of endotoxins on human monocytes. J Biol Chem. 1991 Oct 15;266(29):19490–19498. [PubMed] [Google Scholar]
  6. Golenbock D. T., Liu Y., Millham F. H., Freeman M. W., Zoeller R. A. Surface expression of human CD14 in Chinese hamster ovary fibroblasts imparts macrophage-like responsiveness to bacterial endotoxin. J Biol Chem. 1993 Oct 15;268(29):22055–22059. [PubMed] [Google Scholar]
  7. Han J., Lee J. D., Tobias P. S., Ulevitch R. J. Endotoxin induces rapid protein tyrosine phosphorylation in 70Z/3 cells expressing CD14. J Biol Chem. 1993 Nov 25;268(33):25009–25014. [PubMed] [Google Scholar]
  8. Haziot A., Chen S., Ferrero E., Low M. G., Silber R., Goyert S. M. The monocyte differentiation antigen, CD14, is anchored to the cell membrane by a phosphatidylinositol linkage. J Immunol. 1988 Jul 15;141(2):547–552. [PubMed] [Google Scholar]
  9. Haziot A., Rong G. W., Silver J., Goyert S. M. Recombinant soluble CD14 mediates the activation of endothelial cells by lipopolysaccharide. J Immunol. 1993 Aug 1;151(3):1500–1507. [PubMed] [Google Scholar]
  10. Hemmi S., Böhni R., Stark G., Di Marco F., Aguet M. A novel member of the interferon receptor family complements functionality of the murine interferon gamma receptor in human cells. Cell. 1994 Mar 11;76(5):803–810. doi: 10.1016/0092-8674(94)90355-7. [DOI] [PubMed] [Google Scholar]
  11. Hemmi S., Peghini P., Metzler M., Merlin G., Dembic Z., Aguet M. Cloning of murine interferon gamma receptor cDNA: expression in human cells mediates high-affinity binding but is not sufficient to confer sensitivity to murine interferon gamma. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9901–9905. doi: 10.1073/pnas.86.24.9901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kirkland T. N., Qureshi N., Takayama K. Diphosphoryl lipid A derived from lipopolysaccharide (LPS) of Rhodopseudomonas sphaeroides inhibits activation of 70Z/3 cells by LPS. Infect Immun. 1991 Jan;59(1):131–136. doi: 10.1128/iai.59.1.131-136.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kitchens R. L., Munford R. S. Enzymatically deacylated lipopolysaccharide (LPS) can antagonize LPS at multiple sites in the LPS recognition pathway. J Biol Chem. 1995 Apr 28;270(17):9904–9910. doi: 10.1074/jbc.270.17.9904. [DOI] [PubMed] [Google Scholar]
  14. Kitchens R. L., Ulevitch R. J., Munford R. S. Lipopolysaccharide (LPS) partial structures inhibit responses to LPS in a human macrophage cell line without inhibiting LPS uptake by a CD14-mediated pathway. J Exp Med. 1992 Aug 1;176(2):485–494. doi: 10.1084/jem.176.2.485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kovach N. L., Yee E., Munford R. S., Raetz C. R., Harlan J. M. Lipid IVA inhibits synthesis and release of tumor necrosis factor induced by lipopolysaccharide in human whole blood ex vivo. J Exp Med. 1990 Jul 1;172(1):77–84. doi: 10.1084/jem.172.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. LeGrand C. B., Thieringer R. CD14-dependent induction of protein tyrosine phosphorylation by lipopolysaccharide in murine B-lymphoma cells. Biochim Biophys Acta. 1994 Aug 11;1223(1):36–46. doi: 10.1016/0167-4889(94)90071-x. [DOI] [PubMed] [Google Scholar]
  17. Lee J. D., Kato K., Tobias P. S., Kirkland T. N., Ulevitch R. J. Transfection of CD14 into 70Z/3 cells dramatically enhances the sensitivity to complexes of lipopolysaccharide (LPS) and LPS binding protein. J Exp Med. 1992 Jun 1;175(6):1697–1705. doi: 10.1084/jem.175.6.1697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lee J. D., Kravchenko V., Kirkland T. N., Han J., Mackman N., Moriarty A., Leturcq D., Tobias P. S., Ulevitch R. J. Glycosyl-phosphatidylinositol-anchored or integral membrane forms of CD14 mediate identical cellular responses to endotoxin. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):9930–9934. doi: 10.1073/pnas.90.21.9930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Libermann T. A., Baltimore D. Activation of interleukin-6 gene expression through the NF-kappa B transcription factor. Mol Cell Biol. 1990 May;10(5):2327–2334. doi: 10.1128/mcb.10.5.2327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lynn W. A., Golenbock D. T. Lipopolysaccharide antagonists. Immunol Today. 1992 Jul;13(7):271–276. doi: 10.1016/0167-5699(92)90009-V. [DOI] [PubMed] [Google Scholar]
  21. Lynn W. A., Raetz C. R., Qureshi N., Golenbock D. T. Lipopolysaccharide-induced stimulation of CD11b/CD18 expression on neutrophils. Evidence of specific receptor-based response and inhibition by lipid A-based antagonists. J Immunol. 1991 Nov 1;147(9):3072–3079. [PubMed] [Google Scholar]
  22. Martin M. A. Epidemiology and clinical impact of gram-negative sepsis. Infect Dis Clin North Am. 1991 Dec;5(4):739–752. [PubMed] [Google Scholar]
  23. Morrison D. C. Bacterial endotoxins and pathogenesis. Rev Infect Dis. 1983 Sep-Oct;5 (Suppl 4):S733–S747. doi: 10.1093/clinids/5.supplement_4.s733. [DOI] [PubMed] [Google Scholar]
  24. Morrison D. C., Ryan J. L. Endotoxins and disease mechanisms. Annu Rev Med. 1987;38:417–432. doi: 10.1146/annurev.me.38.020187.002221. [DOI] [PubMed] [Google Scholar]
  25. Pugin J., Heumann I. D., Tomasz A., Kravchenko V. V., Akamatsu Y., Nishijima M., Glauser M. P., Tobias P. S., Ulevitch R. J. CD14 is a pattern recognition receptor. Immunity. 1994 Sep;1(6):509–516. doi: 10.1016/1074-7613(94)90093-0. [DOI] [PubMed] [Google Scholar]
  26. Pugin J., Schürer-Maly C. C., Leturcq D., Moriarty A., Ulevitch R. J., Tobias P. S. Lipopolysaccharide activation of human endothelial and epithelial cells is mediated by lipopolysaccharide-binding protein and soluble CD14. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2744–2748. doi: 10.1073/pnas.90.7.2744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Setoguchi M., Nasu N., Yoshida S., Higuchi Y., Akizuki S., Yamamoto S. Mouse and human CD14 (myeloid cell-specific leucine-rich glycoprotein) primary structure deduced from cDNA clones. Biochim Biophys Acta. 1989 Jul 7;1008(2):213–222. doi: 10.1016/0167-4781(80)90012-3. [DOI] [PubMed] [Google Scholar]
  28. Sibley C. H., Terry A., Raetz C. R. Induction of kappa light chain synthesis in 70Z/3 B lymphoma cells by chemically defined lipid A precursors. J Biol Chem. 1988 Apr 15;263(11):5098–5103. [PubMed] [Google Scholar]
  29. Takayama K., Qureshi N., Beutler B., Kirkland T. N. Diphosphoryl lipid A from Rhodopseudomonas sphaeroides ATCC 17023 blocks induction of cachectin in macrophages by lipopolysaccharide. Infect Immun. 1989 Apr;57(4):1336–1338. doi: 10.1128/iai.57.4.1336-1338.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ulevitch R. J., Tobias P. S. Recognition of endotoxin by cells leading to transmembrane signaling. Curr Opin Immunol. 1994 Feb;6(1):125–130. doi: 10.1016/0952-7915(94)90043-4. [DOI] [PubMed] [Google Scholar]
  31. Wright S. D., Ramos R. A., Tobias P. S., Ulevitch R. J., Mathison J. C. CD14, a receptor for complexes of lipopolysaccharide (LPS) and LPS binding protein. Science. 1990 Sep 21;249(4975):1431–1433. doi: 10.1126/science.1698311. [DOI] [PubMed] [Google Scholar]
  32. Zhang Y., Doerfler M., Lee T. C., Guillemin B., Rom W. N. Mechanisms of stimulation of interleukin-1 beta and tumor necrosis factor-alpha by Mycobacterium tuberculosis components. J Clin Invest. 1993 May;91(5):2076–2083. doi: 10.1172/JCI116430. [DOI] [PMC free article] [PubMed] [Google Scholar]

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