TransgenicEimeria magnaPérard, 1925 Displays Similar Parasitological Properties to the Wild-type Strain and Induces an Exogenous Protein-Specific Immune Response in Rabbits (Oryctolagus cuniculusL.).
Rabbit coccidiosis causes great economic losses to world rabbitries. Little work has been done considering genetic manipulation on the etiological agents, rabbitEimeriaspp. In this study, we constructed a transgenic line ofEimeria magna(EmagER) expressing enhanced yellow fluorescent protein (EYFP) and red fluorescent protein (RFP) using regulatory sequences ofEimeria tenellaandToxoplasma gondii. We observed the life cycle ofEmagERand confirmed that the transgenic parasites express exogenous proteins targeted to different cellular compartments throughout the entire life cycle. EYFP was expressed mainly in the nucleus and RFP both in the nucleus and cytoplasm. Then, coccidia-free, laboratory-reared 40-day-old rabbits were primarily infected with eitherEmagERor wild-type strain oocysts and challenged with the wild-type strain.EmagERshowed similar reproductivity and immunogenicity to the wild-type strain. Finally, we examined the foreign protein-specific immune response elicited byEmagER. Rabbits were immunized with either transgenic or wild-type oocysts. Immune response against parasite-soluble antigen, EYFP and RFP in spleen, and mesenteric lymph nodes were detected by quantitative real-time PCR. The relative expression level of IFN-γ, IL-2, and TNF-α were higher inEmagER-immunized rabbits than wild-type parasites-immunized rabbits after stimulation with EYFP and RFP. Our study confirmed that a specific immune response was induced by the exogenous protein expressed byEmagERand favored future studies on application of transgenic rabbit coccidia as recombinant vaccine vectors.
State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, China; National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China.
Geru
G
Tao
Department of Animal Parasitology, Institute of Animal Husbandry and Veterinary Medicine, Zhejiang Academy of Agricultural Science , Hangzhou , China.
Tuanyuan
T
Shi
State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, China; National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China.
Xinming
X
Tang
Department of Biology, University of New Mexico , Albuquerque, NM , USA.
Donald W
DW
Duszynski
State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, China; National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China.
Yunzhou
Y
Wang
State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, China; National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China.
Chao
C
Li
State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, China; National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China.
Jingxia
J
Suo
State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, China; National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China.
Xiuling
X
Tian
State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, China; National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China.
Xianyong
X
Liu
State Key Laboratory of Agrobiotechnology, China Agricultural University, Beijing, China; National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, China.
Xun
X
Suo
Eimeria magna; immunity; life cycle; rabbit coccidia; transgenic coccidia