Recombinant Chicken Interferon-Gamma-Mediated Inhibition of Eimeria tenella Development in vitro and Reduction of Oocyst Production and Body Weight Loss Following Eimeria acervulina Challenge Infection

• Published in: Avian diseases 1998-04, Vol.42 (2), p.307-314
• Main Authors: Lillehoj, H. S., Choi, K. D.
• Format: Article
• Language: English
• Subjects: Animals; Blotting, Western - veterinary; Body weight; Cell Line; Chickens; Chickens - parasitology; Cloning, Molecular; Coccidiosis; Coccidiosis - parasitology; Coccidiosis - prevention & control; Coccidiosis - veterinary; Complementary DNA; Eimeria - drug effects; Eimeria - immunology; Eimeria acervulina; Eimeria tenella; Eimeria tenella - drug effects; Eimeria tenella - growth & development; Eimeria tenella - immunology; Immunity; Infections; Interferon-gamma - chemistry; Interferon-gamma - pharmacology; Interferon-gamma - therapeutic use; Oocysts; Parasites; Poultry Diseases - immunology; Poultry Diseases - parasitology; Poultry Diseases - prevention & control; Recombinant Proteins; Sporozoites; T lymphocytes; Transfection; Weight Loss - drug effects; Weight Loss - immunology
• ISSN: 0005-2086; 1938-4351
• DOI: 10.2307/1592481

Recombinant chicken interferon-gamma (chIFN-γ) was produced in CHO-K1 or Spodoptera frugiperda (SF9) insect cells by transfection with a pcDNA vector or recombinant baculovirus (SF9-interferon-gamma [IFN-γ]) carrying the chIFN-γ gene. A rabbit antibody against a synthetic peptide corresponding to an immunogenic portion of chIFN-γ recognized a 22-23-kDa band in SF9-IFN-γ cell extracts by western blot analysis. Biological activity of recombinant chIFN-γ was shown by its inhibition of vesicular stomatitis virus-induced cytotoxicity of chicken embryonic fibroblast cells in vitro. To investigate the role of chIFN-γ during Eimeria infection, CHCC-OU2 chicken cells either pretreated with chIFN-γ or stably transfected with the chIFN-γ gene were infected with Eimeria tenella sporozoites. IFN-γ demonstrated significant reductions in intracellular sporozoite development without affecting sporozoite invasion of host cells. Furthermore, chickens treated with recombinant chIFN-γ showed decreased oocyst production and significant improvement in body weight gain following Eimeria acervulina challenge infection. These results provide the first direct evidence that chIFN-γ exerts an inhibitory effect against Eimeria and provides a rational basis for use of this cytokine as a vaccine adjuvant against coccidiosis. /// Se produjo un interferón gamma recombinante de pollo (chIFN-γ) en células CHO-K1 o células de insecto Spodoptera frugiperda (SF9) por medio de transfección con el vector pcDNA ó baculovirus recombinante (SF9-interferón gamma [IFN-γ]) que tiene el gen chIFN-γ. Un anticuerpo de conejo contra un péptido sintético correspondiente a una porción inmunogénica de chIFN-γ reconocieron una banda de 22-23 kilodaltons en extractos de células SF9-IFN-γ mediante el análisis de hibridación puntual western. La actividad biológica del recombinante chIFN-g fue demostrada in vitro por la inhibición de la citotoxicidad inducida por el virus de la estomatitis vesicular en fibroblastos de embrión de pollo. Para investigar el papel de chIFN-γ durante la infección con Eimeria, células de pollo CHCC-OU2 pretratadas con chIFN-γ ó transfectadas de forma estable con el gen chIFN-γ se infectaron con esporozoitos de Eimeria tenella. El IFN-γ produjo reducción significante en el desarrollo de esporozoitos intracelulares sin afectar la invasión de la células del huésped por los esporozoitos. Además, los pollos tratados con el recombinante chITF-γ mostraron disminución en la producción de oocis