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Immunogenicity and protective efficacy of solubilized merozoite-enriched Theileria sergenti immunogens. I: Protection against homologous stabilate challenge
B K Baek,I H Choi,B S Kim,R Hansen,* and I Kakoma*
College of Veterinary Medicine, Chonbuk National University, Chonju 560-756, Korea.
Abstract
Theileria sergenti were isolated from infected erythrocytes by hypotonic lysis, and soluble merozoite antigens were purified by sonication and differential centrifugation. The preparation contained 29, 34, 35 and 105 kD immuno-dominant poly-peptides. The soluble antigens (0.5 mg/ml) were prepared and fortified with Freund's adjuvant. Five month old naive Korean calves were subcutaneously inoculated with the preparation and a booster dose was administered 4 weeks later. Nine weeks after the booster dose, vaccinates and controls were challenged with a homologous stabilate (5.6 × 106 RBC/dose, 40% Parasitemia). All animals were monitored for hematocrit, total erythrocyte count, parasitemia and for the specific antibody by Western immunoblot (WB) and indirect immuno-fluorescent antibody (IFA) test. By 18 weeks after vaccination (6 weeks after the challenge), vaccinated cattle had an average IFA titer of 1:10,240 compared with 1:1,280 of the controls. The vaccinates showed negligible change in hematocrit and total RBC count whereas control animals showed significant (p < 0.05) hematological changes and associated anemia. After vaccination and challenge, the antibody responses demonstrated that vaccination had induced significant production of antibody to the 29 and 35 kD polypeptides. The latter polypeptide was much more strongly recognized by the vaccinated animals, and thus it may be a potential candidate for the vaccine.
Figures
Fig. 1 SDS-PAGE results on T. sergenti antigen (Coomassie stain).
Fig. 2 Western immunoblot analysis of T. sergenti antigens.
Key : PO = Positive serum from a cow experimentally infected with T. sergenti., NE = Normal pre-infection serum
Fig. 3 Intensity of immunfluorescence with immune anti-T. sergenti serum (insert shows background for negative bovine serum).
Fig. 4 IFA response in vaccinated(-▪-) and control (-•-) animals.
Fig. 5 Total RBC(-) and HT(--) profile in vaccinated (-▪-) and control (-•-) animals.
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