Author/Authors :
JIA, Lijun Dept. of Veterinary Medicine - Agricultural College of Yanbian University, Yanji, China , GUO, Huanping National Research Center for Protozoan Diseases - Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan , LIU, Mingming National Research Center for Protozoan Diseases - Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan , GAO, Yang National Research Center for Protozoan Diseases - Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan , ZHANG, Lei Dept. of Veterinary Medicine - Agricultural College of Yanbian University, Yanji, China , LI, Hang Dept. of Veterinary Medicine - Agricultural College of Yanbian University, Yanji, China , XIE, Suzhu Dept. of Veterinary Medicine - Agricultural College of Yanbian University, Yanji, China , ZHANG, Ningning Dept. of Veterinary Medicine - Agricultural College of Yanbian University, Yanji, China
Abstract :
Background: We aimed to construct an adenovirus expressing a cross-reactive fragment
of the apical membrane antigen 1 (AMA1) antigen and evaluated the concomitant
immune response in BABL/c mice, allowing protection against N. caninum and T. gondii
infection.
Methods: The study was conducted in Agricultural College of Yanbian University, Yanji,
Jilin, China In 2015-2016. Primers were designed using the AMA1 gene sequences of
N. caninum (AB265823.1) and T. gondii (AF010264.1). After linearization of the plasmid
ADV4-NcAMA1 and the framework plasmid pacAd5, a total of 293T cells were cotransfected
and Ad5-NcAMA1 recombinant adenovirus were packed. BALB/c mice
were inoculated. Simultaneously serum IgG antibody levels and IFN-γand IL-4 cytokine
levels were determined by ELISA. After immunization three times in two weeks, each
group of BABL/c mice were divided into two groups, respectively given intraperitoneal
inoculation by the Neospora tachyzoite and Toxoplasma tachyzoite. Then we observed the
clinical symptoms and statistical survival rate of mice.
Results: The level of IgG in BABL/c mice immunized with Ad5-NcAMA1 was significantly
increased when compared with that of pVAX1-NcAMA1 and PBS groups
(P<0.01). At the same time, the cytokine levels of IFN-γ and IL-4 were also higher in
the Ad4-NcAMA1 group than in the control groups (P<0.01). Moreover, BABL/c mice
immunized with Ad5-NcAMA1, pVAX1-NcAMA1, and PBS showed survival rates of
75%, 45% and 20% after N. caninum infection, and 45%, 10% and 0% after T. gondii
infection, respectively.
Conclusion: The adenovirus vaccineAd5-NcAMA1 could provide protective immunity
against N. caninum and T. gondii infection.
Keywords :
Neospora caninum , Toxoplasma gondii , AMA1 gene , Recombinant adenovirus , Immune response
