Analysis of interferon gamma protein expression in zebrafish (Danio rerio).
Sohye Yoon, Ayham Alnabulsi, Ting Yu Wang, Po Tsang Lee, Tzong-Yueh Chen, Steve Bird, Jun Zou, Christopher J Secombes
Author Information
Sohye Yoon: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK. Electronic address: sohyeyoon@abdn.ac.uk.
Ting Yu Wang: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK.
Po Tsang Lee: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK.
Tzong-Yueh Chen: Laboratory of Molecular Genetics, College of Bioscience and Biotechnology, National Cheng Kung University, Taiwan.
Steve Bird: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK; Molecular Genetics, Department of Biological Sciences, University of Waikato, Hamilton, New Zealand.
Jun Zou: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK.
Christopher J Secombes: Scottish Fish Immunology Research Centre, School of Biological Sciences, University of Aberdeen, UK. Electronic address: c.secombes@abdn.ac.uk.
IFN-γ is a major effector cytokine, produced to induce type I immune responses. It has been cloned in several fish species including zebrafish, however to date few studies have looked at IFN-γ protein expression and bioactivity in fish. Hence, the current study focused on developing a monoclonal antibody (moAb) against zfIFN-γ. We show that the zfIFN-γ moAb specifically recognises E. coli produced recombinant IFN-γ protein and zfIFN-γ produced in transfected HEK293 cells, by Western blot analysis. Next we analysed the production of the native protein following expression induced by PHA stimulation of leukocytes in vitro or antigen re-stimulation in vivo. We show the IFN-γ protein is produced as a dimer, and that a good correlation exists between transcript expression levels and protein levels.
