A versatile, high through-put, bead-based phagocytosis assay for Plasmodium falciparum

Sci Rep. 2017 Oct 31;7(1):14705. doi: 10.1038/s41598-017-13900-4.

Abstract

Antibody-mediated phagocytosis is an important immune effector mechanism against Plasmodium falciparum-infected erythrocytes (IE); however, current phagocytosis assays use IE collected from infected individuals or from in vitro cultures of P. falciparum, making them prone to high variation. A simple, high-throughput flow cytometric assay was developed that uses THP-1 cells and fluorescent beads covalently-coupled with the malarial antigen VAR2CSA. The assay is highly repeatable, provides both the overall percent phagocytosis and semi-quantitates the number of antigen-coupled beads internalized.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antibodies, Protozoan / metabolism
  • Antigens, Protozoan / metabolism
  • Erythrocytes / parasitology
  • Erythrocytes / physiology*
  • Female
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • High-Throughput Screening Assays / methods*
  • Humans
  • Malaria, Falciparum / diagnosis*
  • Male
  • Microspheres
  • Monocytes / parasitology
  • Monocytes / physiology*
  • Phagocytosis
  • Plasmodium falciparum / physiology*
  • Reproducibility of Results
  • THP-1 Cells

Substances

  • Antibodies, Protozoan
  • Antigens, Protozoan
  • VAR2CSA protein, Plasmodium falciparum