TY - JOUR
T1 - Analysis of interferon gamma protein expression in zebrafish (Danio rerio)
AU - Yoon, Sohye
AU - Alnabulsi, Ayham
AU - Wang, Ting Yu
AU - Lee, Po Tsang
AU - Chen, Tzong Yueh
AU - Bird, Steve
AU - Zou, Jun
AU - Secombes, Christopher J.
N1 - Funding Information:
SY was supported by the Scottish Overseas Research Scholarship Awards Scheme (SORSAS) to undertake a PhD program at the University of Aberdeen. PTL was supported by a PhD studentship from the Ministry of Education, Republic of China (Taiwan). TYW was supported by grants from the Ministry of Science and Technology, Taiwan (NSC 100-2313-B-006-002-MY3 and MOST 103-2313-B-006-004-MY3 ) to visit Aberdeen. This research was also funded by the European Commission under the 7th Framework Programme for Research and Technological Development (FP7) of the European Union (grant agreement No. 311993 TARGETFISH).
Publisher Copyright:
© 2016 Elsevier Ltd
PY - 2016/10/1
Y1 - 2016/10/1
N2 - 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.
AB - 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.
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U2 - 10.1016/j.fsi.2016.08.023
DO - 10.1016/j.fsi.2016.08.023
M3 - Article
C2 - 27539703
AN - SCOPUS:84983520381
VL - 57
SP - 79
EP - 86
JO - Fish and Shellfish Immunology
JF - Fish and Shellfish Immunology
SN - 1050-4648
ER -