title: Transgenic Eimeria tenella expressing enhanced yellow fluorescent protein targeted to different cellular compartments stimulated dichotomic immune responses in chickens.
abstract: Eimeria tenella, one of the seven species of chicken coccidia, elicits protective immunity against challenge infection with both homologous and heterologous strains. We endeavor to use recombinant E. tenella as a vaccine vehicle for expressing and delivering pathogen Ags and investigate immune responses against these foreign Ags. In this study, two lines of transgenic E. tenella expressing a model Ag, enhanced yellow fluorescent protein (EYFP), targeted to the micronemes and to the cytoplasm of the recombinant parasites were constructed to study the impact of Ag compartmentalization on immunogenicity. The MTT assay, intracellular cytokine staining, and real-time PCR were performed to detect the EYFP-specific proliferation and effector functions of splenic lymphocytes of immunized chickens. ELISA was used to measure anti-EYFP IgG and IgA responses. The results showed that both lines of transgenic parasites stimulated EYFP-specific lymphocyte proliferation and IFN-γ expression in CD4 and CD8 T cells, whereas a higher level of Ag-specific lymphocyte proliferation was elicited by the transgenic line expressing microneme-targeted EYFP. Furthermore, this line stimulated stronger IgA response than the one expressing cytoplasm-targeted EYFP after the second immunization. Our findings are encouraging for further investigation of the effect of Ag compartmentalization in transgenic Eimeria on immunogenicity and for the development of a eukaryotic vaccine vector using genetically modified Apicomplexa parasites.
authors.affiliation: National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China;
authors.forename: Xiaoxi
authors.initials: X
authors.lastname: Huang
authors.forename: Jun
authors.initials: J
authors.lastname: Zou
authors.forename: Hanqian
authors.initials: H
authors.lastname: Xu
authors.forename: Ye
authors.initials: Y
authors.lastname: Ding
authors.forename: Guangwen
authors.initials: G
authors.lastname: Yin
authors.forename: Xianyong
authors.initials: X
authors.lastname: Liu
authors.forename: Xun
authors.initials: X
authors.lastname: Suo
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology; D000953:Antigens, Protozoan / Q000276:immunology; D001426:Bacterial Proteins / Q000276:immunology*; D002645:Chickens / Q000276:immunology* / Q000469:parasitology*; D003048:Coccidiosis / Q000276:immunology / Q000382:microbiology / Q000662:veterinary*; D016786:Eimeria tenella / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D008164:Luminescent Proteins / Q000276:immunology*; D016253:Microscopy, Immunoelectron; D011201:Poultry Diseases / Q000276:immunology* / Q000469:parasitology; D016052:Protozoan Vaccines / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D001426:Bacterial Proteins; D008164:Luminescent Proteins; D016052:Protozoan Vaccines; D011994:Recombinant Proteins; C054562:yellow fluorescent protein, Bacteria