Evaluation of recombinant Neospora caninum antigens purified from silkworm larvae for the protection of N. caninum infection in mice

Mai Yoshimoto; Takahiro Otsuki; Kohei Itagaki; Tatsuya Kato; Tetsuya Kohsaka; Yumino Matsumoto; Kazunori Ike; Enoch Y Park

doi:10.1016/j.jbiosc.2015.04.002

Evaluation of recombinant Neospora caninum antigens purified from silkworm larvae for the protection of N. caninum infection in mice

J Biosci Bioeng. 2015 Dec;120(6):715-9. doi: 10.1016/j.jbiosc.2015.04.002. Epub 2015 Apr 29.

Authors

Mai Yoshimoto¹, Takahiro Otsuki¹, Kohei Itagaki¹, Tatsuya Kato², Tetsuya Kohsaka³, Yumino Matsumoto⁴, Kazunori Ike⁴, Enoch Y Park⁵

Affiliations

¹ Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
² Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Laboratory of Biotechnology, Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
³ Laboratory of Animal Reproduction & Physiology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, Suruga-ku, Shizuoka 422-8529, Japan.
⁴ Laboratory of Veterinary Parasitology, Nippon Veterinary and Life University, Musashino, Tokyo 180-8602, Japan.
⁵ Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan; Laboratory of Biotechnology, Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan. Electronic address: park.enoch@shizuoka.ac.jp.

PMID: 25935502
DOI: 10.1016/j.jbiosc.2015.04.002

Abstract

Three antigens (NcSAG1, NcSRS2 and NcMIC3) from Neospora caninum were expressed using the BmNPV bacmid system in silkworm larvae and purified from the hemolymph. From 20 silkworm larvae, 1.5, 1.2 and 1.4 mg of purified recombinant NcSAG1, NcSRS2 and NcMIC3 were obtained, respectively. When each purified recombinant antigen was immunized with Freund's incomplete adjuvant (FIA) to mice, recombinant NcSAG1 induced a Th2 immune response in immunized mice and produced a SAG1-specific antibody. In the experiment where NcSAG1-immunized mice were challenged with N. caninum, the cerebral N. caninum burden was significantly reduced compared with that of either the FIA- or PBS-immunized mice. Recombinant NcSRS2 or NcMIC3 induced both Th1 and Th2 immune responses, but NcMIC3-immunization did not induce significant production of NcMIC3-specific antibodies. These results suggest that the silkworm can produce recombinant antigens of N. caninum, which can be used as a recombinant vaccine against N. caninum.

Keywords: Antigen; BmNPV bacmid; Neospora caninum; Silkworm; Subunit vaccine.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antigens, Protozoan / genetics
Antigens, Protozoan / immunology*
Antigens, Protozoan / isolation & purification*
Bombyx
Coccidiosis / immunology*
Coccidiosis / prevention & control*
Female
Hemolymph / chemistry
Immunization
Larva
Mice
Mice, Inbred BALB C
Neospora / chemistry*
Neospora / genetics
Neospora / immunology*
Protozoan Vaccines / chemistry
Protozoan Vaccines / immunology
Recombinant Proteins / isolation & purification
Th1 Cells / immunology
Th2 Cells / immunology
Vaccines, Subunit / chemistry
Vaccines, Subunit / immunology

Substances

Antigens, Protozoan
Protozoan Vaccines
Recombinant Proteins
Vaccines, Subunit