Abstract
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.
Highlights
Interleukin (IL)-4 and IL-10 are considered as important Th2 cytokines that promote growth and differentiation for a variety of immune cells such as T cells, NK cells, B cells [1,2], and augments antibody-mediated immune response against extracellular bacteria and parasites
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
The present study demonstrated that biologically active horse IL-4 and IL-10 could be produced in E. coli
Summary
Interleukin (IL)-4 and IL-10 are considered as important Th2 cytokines that promote growth and differentiation for a variety of immune cells such as T cells, NK cells, B cells [1,2], and augments antibody-mediated immune response against extracellular bacteria and parasites. IL-4 promotes growth and differentiation of B-cells, tissue repair, maintains allergic responses and homeostasis [3,4,5]. IL-10 is a multifunctional cytokine produced by a variety of cell types including Th2 cells, dendritic cells, activated macrophages, B cells, and mast cells [6,7]. Recombinant version of horse IL-4 and IL-10 has not been investigated to understand their immunomodulating activities. Immune-modulating activities of recombinant horse IL-4 and IL-10 were investigated in peripheral blood mononuclear cells (PBMCs)
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