Analysis of interferon gamma protein expression in zebrafish (Danio rerio)

IFN-gamma 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-gamma protein expression and bioactivity in fish. Hence, the current study focused on developing a monocl...

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Bibliographic Details
Published in:Fish & Shellfish Immunology
Main Authors: Yoon, Sohye, Alnabulsi, Ayham, Wang, Ting Yu, Lee, Po Tsang, Chen, Tzong-Yueh, Bird, Steve, Zou, Jun, Secombes, Christopher J.
Format: Article in Journal/Newspaper
Language:English
Published: Academic Press 2016
Subjects:
Trout Oncorhynchus-Mykiss
Salmon Salmo-Salar
Ginbuna Crucian Carp
Ifn-Gamma
Molecular Characterization
Takifugu-Rubripes
Immune-Responses
Gene-Expression
T-Bet
Identification
Salmo salar
Online Access:https://espace.library.uq.edu.au/view/UQ:97e9d3d
Description
Summary:IFN-gamma 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-gamma protein expression and bioactivity in fish. Hence, the current study focused on developing a monoclonal antibody (moAb) against zfiFN-gamma. We show that the zfIFN-gamma moAb specifically recognises E. coli produced recombinant IFN-gamma protein and zfIFN-gamma 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-gamma protein is produced as a dimer, and that a good correlation exists between transcript expression levels and protein levels. (C) 2016 Elsevier Ltd. All rights reserved.

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