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Related Experiment Videos

Lipopolysaccharide complexed with soluble CD14 binds to normal human monocytes

C Blondin1, A Le Dur, B Cholley

  • 1INSERM U430, Hôpital Broussais, Paris, France.

European Journal of Immunology
|February 17, 1998
PubMed
Summary

Neisseria meningitidis lipopolysaccharide (LPS) binding to human monocytes involves both membrane CD14 and soluble CD14 (sCD14) from serum. Soluble CD14 facilitates LPS binding to monocytes via an unknown molecule, not just as a shuttle to membrane CD14.

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Area of Science:

  • Immunology
  • Cell Biology
  • Microbiology

Background:

  • Neisseria meningitidis lipopolysaccharide (LPS) is a key component of the bacterial outer membrane.
  • CD14 is a cell surface receptor involved in LPS recognition by immune cells.
  • Soluble CD14 (sCD14) is present in serum and its role in LPS binding is not fully understood.

Purpose of the Study:

  • To compare the binding of fluorescein isothiocyanate-labeled LPS (FITC-LPS) to human monocytes and CD14-transfected Chinese hamster ovary (CHO) cells.
  • To investigate the role of membrane CD14 and soluble CD14 (sCD14) in LPS binding to monocytes.

Main Methods:

  • Flow cytometry was used to quantify FITC-LPS binding.
  • Monocytes and hCD14-CHO cells were treated with anti-CD14 monoclonal antibodies (mAbs) or phosphatidylinositol phospholipase C (PI-PLC) to modulate CD14 expression.

Related Experiment Videos

  • Two-color flow cytometry was employed to study the interaction of FITC-LPS with sCD14 on monocytes.

    • Main Results:

    • FITC-LPS binding to both cell types was dose-dependent, saturable, and enhanced by serum.
    • Monoclonal antibody My4 against CD14 partially inhibited LPS binding to monocytes but completely inhibited binding to hCD14-CHO cells.
    • Removal of membrane CD14 by PI-PLC partially decreased LPS binding to monocytes but totally inhibited it on hCD14-CHO cells.
    • FITC-LPS was found to bind to monocytes in association with sCD14 present in serum.
    • The binding of FITC-LPS/sCD14 complexes was inhibited by anti-CD14 mAb 10G33, suggesting involvement of sCD14 or a different epitope.

    Conclusions:

    • Monocyte binding of LPS is mediated by both membrane CD14 and sCD14.
    • sCD14 facilitates LPS binding to monocytes through an unidentified molecule, not solely acting as a shuttle for membrane CD14.
    • These findings elucidate a more complex mechanism of LPS-monocyte interaction involving soluble factors.