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MICROBIAL IMMUNOLOGY

High-Level Expression of Truncated Surface Antigen P50 of Babesia gibsoni in Insect Cells by Baculovirus and Evaluation of Its Immunogenicity and Antigenicity

Shinya Fukumoto, Xuenan Xuan, Kimie Kadota, Ikuo Igarashi, Chihiro Sugimoto, Kozo Fujisaki, Hideyuki Nagasawa, Takeshi Mikami, Hiroshi Suzuki
Shinya Fukumoto
1National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555
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Xuenan Xuan
1National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555
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Kimie Kadota
1National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555
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Ikuo Igarashi
1National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555
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Chihiro Sugimoto
1National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555
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Kozo Fujisaki
1National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555
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Hideyuki Nagasawa
1National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555
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Takeshi Mikami
2Laboratory of Veterinary Public Health, College of Bioresource Science, Nihon University, Fujisawa, Kanagawa 252-8510, Japan
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Hiroshi Suzuki
1National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555
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  • For correspondence: hisuzuki@obihiro.ac.jp
DOI: 10.1128/CDLI.10.4.596-601.2003
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ABSTRACT

Previously, we identified an immunodominant antigen, P50 of Babesia gibsoni. In the present study, the gene encoding the truncated P50 (rP50t) without a C-terminal hydrophobic region (29 amino acids [aa]) was expressed in insect cells by a recombinant baculovirus. The highly hydrophobic C-terminal 20-aa regions seems to be a transmembrane region, which was evidenced by the fact that rP50t was effectively secreted into the supernatant of insect cells infected with the recombinant baculovirus. N-terminal amino acid sequence analysis of rP50t indicated that N-terminal 19 aa function as a signal peptide. The expression level of rP50t reached up to 2 mg per 108 cells infected with the recombinant baculovirus. The immunogenic property of rP50t was evaluated by an immunization test in mice. Mice immunized with rP50t induced a high-level antibody titer against the B. gibsoni merozoite. Monoclonal antibodies (MAbs) to rP50t were produced in mice to determine the immunogenic regions of P50. The epitope(s) recognized by all five MAbs were located between aa 190 and 273, suggesting that the central part of P50 is a highly immunogenic region. The diagnostic potential of rP50t was evaluated using an enzyme-linked immunosorbent assay (ELISA). The ELISA was able to differentiate clearly (P < 0.0001) between B. gibsoni-infected dog serum and B. canis-infected dog serum or noninfected dog serum. Our results indicated that the rP50t may provide a useful potential immunogenic reagent for use in diagnosis and as a subunit vaccine to control B. gibsoni infection in dogs.

  • Copyright © 2003 American Society for Microbiology
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High-Level Expression of Truncated Surface Antigen P50 of Babesia gibsoni in Insect Cells by Baculovirus and Evaluation of Its Immunogenicity and Antigenicity
Shinya Fukumoto, Xuenan Xuan, Kimie Kadota, Ikuo Igarashi, Chihiro Sugimoto, Kozo Fujisaki, Hideyuki Nagasawa, Takeshi Mikami, Hiroshi Suzuki
Clinical and Diagnostic Laboratory Immunology Jul 2003, 10 (4) 596-601; DOI: 10.1128/CDLI.10.4.596-601.2003

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High-Level Expression of Truncated Surface Antigen P50 of Babesia gibsoni in Insect Cells by Baculovirus and Evaluation of Its Immunogenicity and Antigenicity
Shinya Fukumoto, Xuenan Xuan, Kimie Kadota, Ikuo Igarashi, Chihiro Sugimoto, Kozo Fujisaki, Hideyuki Nagasawa, Takeshi Mikami, Hiroshi Suzuki
Clinical and Diagnostic Laboratory Immunology Jul 2003, 10 (4) 596-601; DOI: 10.1128/CDLI.10.4.596-601.2003
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