Vet World Vol.12 April-2019 Article-4
Research Article
Veterinary World, 12(4): 496-503
https://doi.org/10.14202/vetworld.2019.496-503
Recombinant horse interleukin-4 and interleukin-10 induced a mixed inflammatory cytokine response in horse peripheral blood mononuclear cells
2. Department of Biotechnology, Chaudhary Devi Lal University, Sirsa, Haryana, India.
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-γ), IL-6, tumor necrosis factor-alpha (TNF-α), and IL-10, while recombinant horse IL-10 induced expression of IL-6, IFN-γ, and TNF-α.
Conclusion: The present study demonstrated that biologically active horse IL-4 and IL-10 could be produced in E. coli. Keywords: cell proliferation, cytokines, enzyme-linked immunosorbent assay, real-time polymerase chain reaction.
Keywords: cell proliferation, cytokines, enzyme-linked immunosorbent assay, real-time polymerase chain reaction.
How to cite this article: Saini S, Singha H, Siwach P, Tripathi BN (2019) Recombinant horse interleukin-4 and interleukin-10 induced a mixed inflammatory cytokine response in horse peripheral blood mononuclear cells, Veterinary World, 12(4): 496-503.
Received: 29-11-2018 Accepted: 20-02-2019 Published online: 03-04-2019
Corresponding author: Harisankar Singha E-mail: hsssankarbty@gmail.com
DOI: 10.14202/vetworld.2019.496-503
Copyright: Saini, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.