ZFIN ID: ZDB-PUB-160820-7
Analysis of interferon gamma protein expression in zebrafish (Danio rerio)
Yoon, S., Alnabulsi, A., Wang, T.Y., Lee, P.T., Chen, T.Y., Bird, S., Zou, J., Secombes, C.J.
Date: 2016
Source: Fish & shellfish immunology   57: 79-86 (Journal)
Registered Authors: Zou, Jun
Keywords: Antigen re-stimulation, IFN-╬│, Monoclonal antibody, PHA, Protein expression, Zebrafish
MeSH Terms:
  • Animals
  • Antibodies, Monoclonal/immunology
  • Cytokines/immunology
  • Escherichia coli/genetics
  • HEK293 Cells
  • Humans
  • Interferon-gamma/genetics*
  • Interferon-gamma/immunology
  • Interferon-gamma/metabolism
  • Leukocytes/immunology
  • Recombinant Proteins/genetics
  • Recombinant Proteins/immunology
  • Recombinant Proteins/metabolism
  • Zebrafish/genetics*
  • Zebrafish/immunology*
  • Zebrafish/metabolism
PubMed: 27539703 Full text @ Fish Shellfish Immunol.
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┬ácells, 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.