Use of mouse macrophage cell lines for in vitro propagation of Toxoplasma gondii RH tachyzoites.
In most laboratories, Toxoplasma gondii is maintained in mice and is studied in vitro using nonlymphoid cell lines or primary mouse macrophages. In this study, three rapidly dividing mouse macrophage cell lines (J774 A.1, P388D1, RAW264.7) were evaluated for their suitability for studying the RH strain of T. gondii. For comparison, tachyzoites were also grown in two slowly dividing epithelial cell types: a rat lung cell line (L2) and a bovine turbinate cell line (BT). Various inocula of T. gondii were added to the above cells and tachyzoites were harvested from the culture supernatants after 2-8 days of infection. The mouse macrophage cell lines supported rapid growth of T. gondii RH allowing up to a 300-fold increase of the inoculum in 2-4 days. L2 and BT supported slower growth of T. gondii (10- to 90-fold increase of inoculum in 5 to 8 days) and, thus, may be more suitable for assessment of host cell-parasite interactions and drug activity. Toxoplasma gondii RH isolated from each of the cell cultures described were able to multiply in all cell types used. Protein profiles of whole tachyzoite isolated from mice or cell cultures and protein profiles of the corresponding soluble and membrane fractions of the intraphagosomal membrane network were similar as seen after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In mice, intraperitoneal injection of 10(6), 10(5), and 10(3) tachyzoites isolated from the cell cultures or from infected mice caused death after 4, 5, and 8 days, respectively, indicating that parasites grown in vitro retained virulence.
Chamberland
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Service d'Infectiologie, Université Laval, Sainte-Foy, Quebec, Canada.
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D000818:Animals; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008264:Macrophages / Q000469:parasitology*; D051379:Mice; D015800:Protozoan Proteins / Q000032:analysis*; D011739:Pyrimethamine / Q000494:pharmacology; D013416:Sulfamerazine / Q000494:pharmacology; D014122:Toxoplasma / Q000032:analysis / Q000187:drug effects / Q000254:growth & development* / Q000472:pathogenicity; D014124:Toxoplasmosis, Animal / Q000469:parasitology; D014774:Virulence
D015800:Protozoan Proteins; D013416:Sulfamerazine; D011739:Pyrimethamine