Flow Cytometry Assay of Plasmodium Falciparum-Specific B-Cell Proportions

Methods Mol Biol. 2022:2470:681-688. doi: 10.1007/978-1-0716-2189-9_51.

Abstract

Detection of P. falciparum-specific subpopulations of B-cells is important for studies of immunity in malaria. This protocol relies on the photostability and protein loading capacity of carboxylated quantum dots to detect a broad range of different P. falciparum-specific B-cells. Infected red blood cell ghosts, obtained by permeabilization of infected cells with Streptolysin O, are coupled with carboxyl quantum dots using N-ethyl-N-dimethylaminopropyl-carbodiimide condensation. Immunophenotyping of P. falciparum-specific B-cells is performed by flow cytometry using Fc-receptor block, quantum dot-infected red blood cell ghost conjugates, and fluorochrome-conjugated anti-human CD19 mouse monoclonal antibodies.

Keywords: B-cells; Flow cytometry; Ghost infected red blood cells; Malaria; Plasmodium falciparum; Quantum dots.

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Erythrocytes
  • Flow Cytometry / methods
  • Immunophenotyping
  • Malaria*
  • Malaria, Falciparum*
  • Mice
  • Plasmodium falciparum

Substances

  • Antibodies, Monoclonal