title: Production of a functional chicken single-chain variable fragment antibody derived from caecal tonsils B lymphocytes against macrogamonts of Eimeria tenella.
abstract: Avian coccidiosis is due to a protozoan intracellular parasite belonging to the genus Eimeria which multiplies in the intestine of the host. In order to identify Eimeria antigens which reflect the natural avian humoral immune response, chicken hybridomas were produced by fusion of myeloma MuH1 with B lymphocytes from Eimeria tenella infected chicken. B lymphocytes used for fusions were isolated from tonsils at the basis of caeca where the parasite develops. One of the clones (G1F5) recognised oocyst antigens and the macrogamont stage of the parasite in ELISA and immunofluorescence assay. A single-chain variable fragment (scFv) antibody was cloned from the light chain variant region (VL) and heavy chain variant region (VH) genes of the hybridoma. This recombinant antibody (scFv G1F5) exhibited antigen binding specificity to oocysts and macrogamonts of E. tenella equivalent to the mAb produced by the clone G1F5. Nucleotide sequence analysis of VL genes from scFv G1F5 compared to the germ-line revealed vestiges of gene conversion. scFv derived from chicken B lymphocytes isolated from the gut-associated lymphoid tissue following experimental infection can reveal specific antigens recognised by the avian immune response.
authors.affiliation: Laboratoire de Virologie et Immunologie Moléculaires, INRA, F-78352 Jouy-en-Josas, France.
authors.forename: Susana
authors.initials: S
authors.lastname: Réfega
authors.forename: Marianne
authors.initials: M
authors.lastname: Cluzeaud
authors.forename: Pierre
authors.initials: P
authors.lastname: Péry
authors.forename: Marie
authors.initials: M
authors.lastname: Labbé
authors.forename: Fabienne
authors.initials: F
authors.lastname: Girard-Misguich
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D001483:Base Sequence; D002432:Cecum / Q000276:immunology; D002645:Chickens / Q000276:immunology*; D003001:Cloning, Molecular; D003048:Coccidiosis / Q000276:immunology / Q000469:parasitology / Q000662:veterinary*; D016786:Eimeria tenella / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005455:Fluorescent Antibody Technique / Q000662:veterinary; D006825:Hybridomas / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000276:immunology*; D008221:Lymphoid Tissue / Q000276:immunology; D008969:Molecular Sequence Data; D011201:Poultry Diseases / Q000276:immunology* / Q000469:parasitology; D016415:Sequence Alignment
chemical_list: D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region