期刊
VETERINARY WORLD
卷 12, 期 4, 页码 496-503出版社
VETERINARY WORLD
DOI: 10.14202/vetworld.2019.496-503
关键词
cell proliferation; cytokines; enzyme-linked immunosorbent assay; real-time polymerase chain reaction
资金
- Science and Engineering Research Board (SERB), Department of Science and Technology, New Delhi, India [SERB/F/3326/2012-2013]
Background and Aim: Interleukin (IL)-4 and IL-10 activate plethora of immune cells and induce the humoral immune response. However, recombinant version of horse IL-4 and IL-10 has not been investigated to understand their immunomodulating activities. This study aimed to produce recombinant horse mature IL-4 and IL-10 in Escherichia coli. Immune-modulating activities of recombinant horse IL-4 and IL-10 were investigated in peripheral blood mononuclear cells (PBMCs). Materials and Methods: Equine PBMCs were stimulated with recombinant IL-4 and IL-10. A proliferation of PBMCs was measured by XTT assay and cytokines induction was measured by enzyme-linked immunosorbent assay and real-time polymerase chain reaction. Results: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis displayed a molecular weight of 15 kDa for IL-4 and 19 kDa for IL-10. Recombinant IL-4 and IL-10 significantly induced cell proliferation at 250 ng/ml. The results demonstrated that IL-4 enhanced expression of interferon-gamma (IFN-gamma), IL-6, tumor necrosis factor-alpha (TNF-alpha), and IL-10, while recombinant horse IL-10 induced expression of IL-6, IFN-gamma, and TNF-alpha. Conclusion: The present study demonstrated that biologically active horse IL-4 and IL-10 could be produced in E. coli.
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