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Clinical and Diagnostic Laboratory Immunology, January 2000, p. 9-13, Vol. 7, No. 1
1071-412X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Opsonization and Phagocytosis of Plasmodium falciparum Merozoites Measured by Flow Cytometry

Lakshmi M. Kumaratilake1,* and Antonio Ferrante1,2

Department of Immunopathology1 and University of Adelaide Department of Paediatrics,2 Women's and Children's Hospital, Adelaide, South Australia 5006, Australia

Received 10 May 1999/Returned for modification 9 August 1999/Accepted 16 September 1999

A flow cytometric phagocytosis assay was established to investigate the role of anti-merozoite antibody, complement, and cytokines on the phagocytosis of Plasmodium falciparum merozoites by human neutrophils. This assay involved allowing fluorescein isothiocyanate-labeled merozoites to interact with phagocytes and analysis of the cells on a FACScan with Lysis II software. To differentiate the proportion of neutrophil surface-bound merozoites from the merozoites ingested by neutrophils, the fluorescence of bound merozoites was quenched by adding trypan blue. The data showed that sera from malaria-immune individuals in the Solomon Islands and Papua New Guinea promoted merozoite engulfment by neutrophils. The cytokines tumor necrosis factor alpha, gamma interferon, granulocyte-macrophage colony-stimulating factor, and interleukin-1beta significantly increased the amount and the rate of merozoite phagocytosis by neutrophils. Optimum merozoite phagocytosis occurred when both cytokines and anti-malarial antibody were present.

* Corresponding author. Mailing address: Department of Immunopathology, Women's and Children's Hospital, North Adelaide, South Australia 5006, Australia. Phone: 61-8-82046637 or 61-8-82047216. Fax: 61-8-8204637031. E-mail: lkumarat{at}medicine.adelaide.edu.au.

Clinical and Diagnostic Laboratory Immunology, January 2000, p. 9-13, Vol. 7, No. 1
1071-412X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.




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