Production of biologically active equine interleukin 12 through expression of p35, p40 and single chain IL-12 in mammalian and baculovirus expression systems

McMonagle, E.L.J. et al. (2001) Production of biologically active equine interleukin 12 through expression of p35, p40 and single chain IL-12 in mammalian and baculovirus expression systems. Equine Veterinary Journal, 33(7), pp. 693-698. (doi:10.2746/042516401776249426)

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Abstract

Interleukin-12 (IL-12) is a key cytokine in the development of cell-mediated immune responses. Bioactive IL-12 is a heterodimeric cytokine composed of disulphide linked p35 and p40 subunits. The aim of this study was to verify biologically activity of the products expressed from equine interleukin-12 (IL-12) p35 and p40 cDNAs and to establish whether equine IL-12 could be expressed as a p35/p40 fusion polypeptide, as has been reported for IL-12a of several mammalian species. We report production of equine IL-12 through expression of p35 and p40 subunits in mammalian and insect cells and of a p35:p40 fusion polypeptide in mammalian cells. Conditioned medium recovered from cultures transiently transfected with constructs encoding equine p35 and p40 subunits or single chain IL-12 enhanced IFN-γ production in cells derived from equine lymph nodes. Preincubation of IFN-γ inducing preparations with anti-p40 monoclonal antibody resulted in a significant decrease in IFN-γ induction capacity. Medium recovered from p35 and p40-expressing baculovirus infected cultures enhanced target cell IFN-γ production and proliferation. Experimental studies in mice and other animals have revealed a therapeutic benefit of IL-12 in cancer, inflammatory and infectious disease and an adjuvant effect in prophylactic regimes. Production of a bioactive species-specific IL-12 is a first step towards an investigation of its potential application in equine species.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Onions, Professor David and McMonagle, Mrs Elizabeth and Nicolson, Dr Lesley
Authors: McMonagle, E.L.J., Taylor, S., van Zuilekom, H., Sanders, L., Scholtes, N., Keanie, L.J., Hopkins, C.A., Logan, N.A., Bain, D., Argyle, D.J., Onions, D.E., Schijns, V.E., and Nicolson, L.
College/School:College of Medical Veterinary and Life Sciences > School of Veterinary Medicine
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Equine Veterinary Journal
Publisher:Blackwell Publishing
ISSN:0425-1644
ISSN (Online):2042-3306
Published Online:05 January 2010

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