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Cloning of African Trypanosomes in Mice Immunosuppressed by Cyclophosphamide Treatment
Chris J. Smith
Chris J. Smith
Department of Microbiology and Immunology, School of Medicine, University of Louisville, Louisville, Kentucky 40292
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Rita F. Levine
Rita F. Levine
Department of Microbiology and Immunology, School of Medicine, University of Louisville, Louisville, Kentucky 40292
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John M. Mansfield
John M. Mansfield
Department of Microbiology and Immunology, School of Medicine, University of Louisville, Louisville, Kentucky 40292
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abstract
Abstract. The use of cyclophosphamide-treated mice for purposes of cloning the African trypanosomes was assessed. C3HeB/FeJ mice were injected with 200 mg/kg cyclophosphamide (CY) 24–72 hours prior to infection with a single trypanosome isolated from Trypanosoma rhodesiense infected blood samples. All CY-treated mice exhibited depressed parasite-specific and antigen nonspecific B lymphocyte responses for a period of 10 days after CY exposure, which was a period of time sufficient to grow trypanosomes from a single organism to a fulminating parasitemia. Cloning efficiency was routinely 45% in these animals. Thus, our study demonstrates that CY-treated mice are a convenient and efficient vehicle for cloning African trypanosomes. Techniques which facilitate the selection of single trypanosomes from infected blood are also described in this report.
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