Multiple oral inoculations with Cryptosporidium parvum as a means of immunization for production of monoclonal antibodies |
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Authors: | Michael Tilley Mark T Eggleston Steve J Upton |
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Institution: | Division of Biology, Kansas State University, Manhattan, KS 66506, USA |
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Abstract: | Abstract Oral immunization of suckling mice with Cryptosporidium parvum results in a humoral response to a limited set of antigens. Six-day-old BALB/c mice were each inoculated orally with 1 × 106 viable oocysts and subsequently administered oral inoculations of 2 × 106 viable oocysts at 30 and 60 days following the primary infection. After 45 days, mice were boosted with 1 × 106 oocysts orally, plus soluble extracts equivalent to 2 × 106 and 1 × 106 oocysts given intravenously and intraperitoneally, respectively. Four days later, splenic lymphocytes were fused to Ag8 myeloma cells. Using this method, we have been able to select for monoclonal antibodies that predominately recognize sporozoite surface and apical complex antigens. |
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Keywords: | Cryptosporidium parvum Coccidia Monoclonal antibody Cell culture Western blotting |
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