Skip to main content
PMC home page
Immunology logoLink to Immunology
. 1993 Sep;80(1):78–83.

Anti-CD14 antibodies reduce responses of cultured human endothelial cells to endotoxin.

E J von Asmuth 1, M A Dentener 1, V Bazil 1, M G Bouma 1, J F Leeuwenberg 1, W A Buurman 1
PMCID: PMC1422125  PMID: 7503948

Abstract

Lipopolysaccharide (LPS) activates both myeloid and endothelial cells. Whereas CD14 has been shown to be involved in LPS recognition by myeloid cells, the mechanism responsible for the strong response of endothelial cells to LPS remains to be elucidated. The role of CD14 in this process was studied using CD14-specific antibodies (Ab). Anti-CD14 Ab inhibited LPS-induced interleukin-6 (IL-6) release and E-selectin expression by cultured human umbilical vein endothelial cells (HUVEC). Messenger RNA encoding IL-6 and E-selectin was reduced in parallel. The inhibitory effect of anti-CD14 Ab was epitope dependent, maximal at low LPS concentrations and dropping with increasing LPS doses. Anti-CD14 Ab did not affect endothelial cell activation induced by IL-1 beta, tumour necrosis factor-alpha (TNF-alpha) and phorbol 12-myristate 13-acetate (PMA). IL-6 release and E-selectin expression of HUVEC were strongly reduced when LPS activation was performed in the absence of serum, indicating involvement of serum components in LPS activation of HUVEC. Nevertheless, anti-CD14 Ab also blocked LPS-induced HUVEC activation in the absence of serum. Although the role of serum components in LPS activation remains to be elucidated, CD14 seems to be a key mediator in LPS-induced activation of endothelial cells.

Full text

PDF

Images in this article

80Figure 1
on p.80

Selected References

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

  1. Bazil V., Horejsí V., Baudys M., Kristofová H., Strominger J. L., Kostka W., Hilgert I. Biochemical characterization of a soluble form of the 53-kDa monocyte surface antigen. Eur J Immunol. 1986 Dec;16(12):1583–1589. doi: 10.1002/eji.1830161218. [DOI] [PubMed] [Google Scholar]
  2. Bevilacqua M. P., Pober J. S., Mendrick D. L., Cotran R. S., Gimbrone M. A., Jr Identification of an inducible endothelial-leukocyte adhesion molecule. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9238–9242. doi: 10.1073/pnas.84.24.9238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bogman M. J., Dooper I. M., van de Winkel J. G., Tax W. J., Hoitsma A. J., Assmann K. J., Ruiter D. J., Koene R. A. Diagnosis of renal allograft rejection by macrophage immunostaining with a CD14 monoclonal antibody, WT14. Lancet. 1989 Jul 29;2(8657):235–238. doi: 10.1016/s0140-6736(89)90427-3. [DOI] [PubMed] [Google Scholar]
  4. Dentener M. A., Bazil V., Von Asmuth E. J., Ceska M., Buurman W. A. Involvement of CD14 in lipopolysaccharide-induced tumor necrosis factor-alpha, IL-6 and IL-8 release by human monocytes and alveolar macrophages. J Immunol. 1993 Apr 1;150(7):2885–2891. [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Heumann D., Gallay P., Barras C., Zaech P., Ulevitch R. J., Tobias P. S., Glauser M. P., Baumgartner J. D. Control of lipopolysaccharide (LPS) binding and LPS-induced tumor necrosis factor secretion in human peripheral blood monocytes. J Immunol. 1992 Jun 1;148(11):3505–3512. [PubMed] [Google Scholar]
  8. Hogg N., MacDonald S., Slusarenko M., Beverley P. C. Monoclonal antibodies specific for human monocytes, granulocytes and endothelium. Immunology. 1984 Dec;53(4):753–767. [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Krüger C., Schütt C., Obertacke U., Joka T., Müller F. E., Knöller J., Köller M., König W., Schönfeld W. Serum CD14 levels in polytraumatized and severely burned patients. Clin Exp Immunol. 1991 Aug;85(2):297–301. doi: 10.1111/j.1365-2249.1991.tb05722.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Labeta M. O., Landmann R., Obrecht J. P., Obrist R. Human B cells express membrane-bound and soluble forms of the CD14 myeloid antigen. Mol Immunol. 1991 Jan-Feb;28(1-2):115–122. doi: 10.1016/0161-5890(91)90094-z. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Leeuwenberg J. F., Jeunhomme G. M., Buurman W. A. Characterization of two monoclonal antibodies directed against an adhesion molecule on human endothelial cells. Transplant Proc. 1990 Aug;22(4):1991–1993. [PubMed] [Google Scholar]
  14. Loppnow H., Libby P. Adult human vascular endothelial cells express the IL6 gene differentially in response to LPS or IL1. Cell Immunol. 1989 Sep;122(2):493–503. doi: 10.1016/0008-8749(89)90095-6. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. May L. T., Torcia G., Cozzolino F., Ray A., Tatter S. B., Santhanam U., Sehgal P. B., Stern D. Interleukin-6 gene expression in human endothelial cells: RNA start sites, multiple IL-6 proteins and inhibition of proliferation. Biochem Biophys Res Commun. 1989 Mar 31;159(3):991–998. doi: 10.1016/0006-291x(89)92206-7. [DOI] [PubMed] [Google Scholar]
  17. Medeiros L. J., Herrington R. D., Gonzalez C. L., Jaffe E. S., Cossman J. My4 antibody staining of non-Hodgkin's lymphomas. Am J Clin Pathol. 1991 Mar;95(3):363–368. doi: 10.1093/ajcp/95.3.363. [DOI] [PubMed] [Google Scholar]
  18. Muller W. A., Ratti C. M., McDonnell S. L., Cohn Z. A. A human endothelial cell-restricted, externally disposed plasmalemmal protein enriched in intercellular junctions. J Exp Med. 1989 Aug 1;170(2):399–414. doi: 10.1084/jem.170.2.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schleimer R. P., Rutledge B. K. Cultured human vascular endothelial cells acquire adhesiveness for neutrophils after stimulation with interleukin 1, endotoxin, and tumor-promoting phorbol diesters. J Immunol. 1986 Jan;136(2):649–654. [PubMed] [Google Scholar]
  20. Simmons D. L., Tan S., Tenen D. G., Nicholson-Weller A., Seed B. Monocyte antigen CD14 is a phospholipid anchored membrane protein. Blood. 1989 Jan;73(1):284–289. [PubMed] [Google Scholar]
  21. Van Voorhis W. C., Steinman R. M., Hair L. S., Luban J., Witmer M. D., Koide S., Cohn Z. A. Specific antimononuclear phagocyte monoclonal antibodies. Application to the purification of dendritic cells and the tissue localization of macrophages. J Exp Med. 1983 Jul 1;158(1):126–145. doi: 10.1084/jem.158.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wright S. D., Ramos R. A., Hermanowski-Vosatka A., Rockwell P., Detmers P. A. Activation of the adhesive capacity of CR3 on neutrophils by endotoxin: dependence on lipopolysaccharide binding protein and CD14. J Exp Med. 1991 May 1;173(5):1281–1286. doi: 10.1084/jem.173.5.1281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wright S. D., Ramos R. A., Patel M., Miller D. S. Septin: a factor in plasma that opsonizes lipopolysaccharide-bearing particles for recognition by CD14 on phagocytes. J Exp Med. 1992 Sep 1;176(3):719–727. doi: 10.1084/jem.176.3.719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Ziegler-Heitbrock H. W., Passlick B., Flieger D. The monoclonal antimonocyte antibody My4 stains B lymphocytes and two distinct monocyte subsets in human peripheral blood. Hybridoma. 1988 Dec;7(6):521–527. doi: 10.1089/hyb.1988.7.521. [DOI] [PubMed] [Google Scholar]
  26. von Asmuth E. J., Leeuwenberg J. F., Ceska M., Buurman W. A. LPS and cytokine-induced endothelial cell IL-6 release and ELAM-1 expression; involvement of serum. Eur Cytokine Netw. 1991 Aug-Sep;2(4):291–297. [PubMed] [Google Scholar]

Articles from Immunology are provided here courtesy of British Society for Immunology

RESOURCES