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Expression and functional characterization of recombinant chicken interferon-gamma
K D Song1, H S Lillehoj, K D Choi
1Department of Animal Science and Technology, College of Agriculture and Life Sciences, Seoul National University, Suweon, South Korea.
Veterinary Immunology and Immunopathology
|January 22, 1998
Summary
Chicken interferon-gamma (IFN-gamma) was cloned and expressed, demonstrating antiviral properties and the ability to regulate macrophage activity. This research provides insights into avian immune responses and potential therapeutic applications.
Area of Science:
- Immunology
- Virology
- Molecular Biology
Background:
- Interferon-gamma (IFN-gamma) is a critical cytokine involved in immune responses.
- Understanding avian IFN-gamma is essential for poultry health and disease control.
Purpose of the Study:
- To clone, express, and characterize chicken interferon-gamma (chIFN-gamma).
- To investigate the antiviral and immunoregulatory functions of chIFN-gamma.
Main Methods:
- Cloning of chIFN-gamma cDNA using reverse transcription-polymerase chain reaction (RT-PCR).
- Expression of recombinant chIFN-gamma in bacterial (E. coli) and eukaryotic (COS, CEC-32) cell lines.
- Analysis of gene expression kinetics via Northern blot and RT-PCR.
- Immunoprecipitation assays to identify protein products.
- In vitro assays to assess antiviral activity against vesicular stomatitis virus (VSV) and macrophage antigen expression.
Main Results:
- Recombinant chIFN-gamma (rchIFN-gamma) exhibited high bioactivity in vitro.
- IFN-gamma mRNA expression peaked at 2 hours post-activation in Con A-stimulated lymphocytes.
- rchIFN-gamma demonstrated significant inhibition of VSV-mediated cytotoxicity in chicken embryonic fibroblast (CEF) cells.
- chIFN-gamma upregulated macrophage cell surface antigens, including MHC class I and II proteins.
Conclusions:
- Chicken interferon-gamma possesses potent antiviral activity.
- chIFN-gamma plays a role in immunoregulating macrophage functions.
- The findings contribute to understanding avian immune mechanisms and potential therapeutic strategies.