Flow cytometric quantification of Toxoplasma gondii cellular infection and replication

The invasion and replication of Toxoplasma gondii are usually analyzed through either optical microscopy or incorporation of tritiated uracil. A new method has been developed using flow cytometric analysis to examine the entry and replication of T. gondii RH strain in Saimiri brain endothelial cells...

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Bibliographic Details
Published inThe Journal of parasitology Vol. 85; no. 3; p. 545
Main Authors Gay-Andrieu, F, Cozon, G J, Ferrandiz, J, Kahi, S, Peyron, F
Format Journal Article
LanguageEnglish
Published United States 01.06.1999
Subjects
Animals
Brain - cytology
Brain - parasitology
Cells, Cultured
Endothelium - cytology
Endothelium - parasitology
Flow Cytometry
Kinetics
Microscopy, Fluorescence
Saimiri
Toxoplasma - physiology
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Summary:The invasion and replication of Toxoplasma gondii are usually analyzed through either optical microscopy or incorporation of tritiated uracil. A new method has been developed using flow cytometric analysis to examine the entry and replication of T. gondii RH strain in Saimiri brain endothelial cells. After cell fixation and permeabilization using saponin, intracellular T. gondii were labeled with a monoclonal antibody against T. gondii SAG-1 (P30; the major cell-surface antigen) followed by fluorescein-conjugated rabbit anti-mouse IgG. The percentage of infected cells obtained using flow cytometry correlated directly with that obtained by UV light microscopy (r = 0.97). The mean fluorescence intensity of infected cells reflects intracellular P30 and assesses intracellular replication. The distribution of fluorescence per infected cell, considered with the percentage of infected cells, also allows a qualitative analysis of replication. Such a method is rapid, easy, and does not require specialized equipment for radioactive labeling.
ISSN:0022-3395
DOI:10.2307/3285793