Recombinant bovine herpesvirus-1 expressing p23 protein of Cryptosporidium parvum induces neutralizing antibodies in rabbits.
In order to develop a vaccine against cryptosporidiosis in cattle, we constructed a recombinant bovine herpesvirus-1 (BHV-1) expressing an immunodominant surface protein, p23, of Cryptosporidium parvum sporozoites. In the recombinant virus, the p23 gene under the control of a CAG promoter and a gene coding for an enhanced green fluorescent protein were integrated into the gG gene of BHV-1. Despite a low frequency of homologous recombination, cloning of the recombinants was easy because of the specific fluorescence of the plaques formed by recombinants. These plaques were among the plaques of the nonfluorescent parental virus. All clones selected for fluorescence also contained the p23 gene. In MDBK cells infected with the recombinant BHV-1, the antibody against the p23 protein recognized the p23 protein as an approximately 23-kDa specific band in Western blotting analysis. Rabbits immunized with the recombinant produced IgG against the p23 protein. It was also demonstrated that the sera of immunized rabbits reduced infection of C. parvum sporozoites in HCT-8 cells. The serum of an immunized rabbit reduced infection compared with the normal rabbit serum control. These results indicate that the recombinant BHV-1 induces neutralizing antibodies in rabbits.
Takashima
Yasuhiro
Y
Department of Global Agricultural Science, Graduate School of Agriculture and Life Science, The University of Tokyo, Tokyo, Japan.
Xuan
Xuenan
X
Kimata
Isao
I
Iseki
Motohiro
M
Kodama
Yoshikatsu
Y
Nagane
Noriko
N
Nagasawa
Hideyuki
H
Matsumoto
Yasunobu
Y
Mikami
Takeshi
T
Otsuka
Haruki
H
D000818:Animals; D000913:Antibodies, Protozoan / Q000096:biosynthesis / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D002417:Cattle; D002460:Cell Line; D003457:Cryptosporidiosis / Q000517:prevention & control*; D016785:Cryptosporidium parvum / Q000276:immunology*; D005260:Female; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D049452:Green Fluorescent Proteins; D007242:Herpesvirus 1, Bovine / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D007106:Immune Sera / Q000276:immunology; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D016052:Protozoan Vaccines; D011817:Rabbits; D014614:Vaccines, Synthetic
D000913:Antibodies, Protozoan; D000914:Antibodies, Viral; D006023:Glycoproteins; D007106:Immune Sera; D008164:Luminescent Proteins; D015800:Protozoan Proteins; D016052:Protozoan Vaccines; D014614:Vaccines, Synthetic; D049452:Green Fluorescent Proteins