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Related Experiment Videos

Quantifying complement-mediated phagocytosis by human monocyte-derived macrophages.

H T Chan1, K Kedzierska, J O'Mullane

  • 1AIDS Pathogenesis Research Unit, Macfarlane Burnet Centre for Medical Research, Fairfield, Victoria, Australia.

Immunology and Cell Biology
|September 21, 2001
PubMed
Summary
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Sheep red blood cells (SRBC) opsonized with human serum trigger complement-mediated phagocytosis by macrophages. HIV-1 infection impairs this crucial immune response, offering a new tool for research.

Area of Science:

  • Immunology
  • Cell Biology
  • Virology

Background:

  • Complement-mediated phagocytosis is essential for immune surveillance.
  • Human immunodeficiency virus type 1 (HIV-1) infects macrophages, key players in the immune system.
  • The impact of HIV-1 on complement receptor-mediated phagocytosis by macrophages remains incompletely understood.

Purpose of the Study:

  • To investigate the role of complement receptors in the phagocytosis of opsonized sheep red blood cells (SRBC) by human monocyte-derived macrophages.
  • To determine if HIV-1 infection affects complement-mediated phagocytosis.
  • To establish a quantitative assay for studying phagocytosis in adherent macrophages.

Main Methods:

  • Opsonization of SRBC with untreated or heat-inactivated human serum.

Related Experiment Videos

  • Quantification of SRBC phagocytosis by human monocyte-derived macrophages using a colorimetric assay.
  • Assessment of the role of complement receptors via inhibition studies with a protein kinase C (PKC)-specific inhibitor (RO 31-8220) and augmentation with phorbol ester (PMA).
  • Evaluation of phagocytosis in macrophages infected with HIV-1.

    • Main Results:

    • SRBC opsonized with untreated human serum were efficiently phagocytosed, with minimal lysis.
    • Phagocytosis was mediated exclusively by complement receptors, as evidenced by the lack of uptake with heat-inactivated serum and augmentation by PMA.
    • Inhibition of protein kinase C (PKC) significantly reduced phagocytosis.
    • HIV-1 infection of macrophages markedly inhibited complement-mediated phagocytosis of SRBC.

    Conclusions:

    • Complement receptors are the primary mediators of SRBC phagocytosis by human monocyte-derived macrophages.
    • HIV-1 infection impairs complement-mediated phagocytosis in these cells.
    • The developed assay is a valuable tool for studying complement-mediated phagocytosis and its modulation by pathogens or pharmacological agents.