Aim: To clone human IL-1Ra gene and express it in E.coli.
Methods: Human IL-1Ra cDNA was obtained by RT-PCR with the the total RNA extracted from human peripheral blood leucocytes as template. The cDNA was cloned into pBV220 vector and expressed. The expressed product was renatured and purified. Separation, purification and bioactivity analysis of the expressed products were performed.
Results: IL-1Ra gene was successfully expressed in E.coli and the expression level reached to about 40% of total bacteria protein. The purity of the final product was over 98%. The product could obviously suppress the secretion of IL-2 by EL-4 cells stimulated with IL-1beta.
Conclusion: The expression of hIL-1Ra gene with bioactivity in E.coli lays experimental foundation for further development and utilisation.
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Immunol Lett
April 2016
Rheumatism Research Center, Catholic Research Institute of Medical Science, The Catholic University of Korea, Seoul 137-040, South Korea; Division of Rheumatology, Department of internal Medicine, School of Medicine, The Catholic University of Korea, Seoul 137-070, South Korea. Electronic address:
Introduction: IL-1β signalling has a critical role in the pathogenesis of various types of inflammatory arthritis including rheumatoid arthritis (RA). We aimed to investigate the therapeutic effects of human IL-1 receptor antagonist with Fc fragment (hIL-1Ra-Fc) on autoimmune arthritis and to identify the possible mechanisms by which hIL-1RA-Fc exerts anti-arthritic effects in a murine model of RA and patients with rheumatoid arthritis.
Methods: Collagen-induced arthritis (CIA) murine model was established in DBA/1J mice.
PLoS One
August 2014
Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, Tennessee, United States of America.
The objective of this study was to determine the potential of human bone marrow derived mesenchymal stem cells (hBMSCs) as gene carriers for improving the outcome of human islet transplantation. hBMSCs were characterized for the expression of phenotypic markers and transduced with Adv-hVEGF-hIL-1Ra to overexpress human vascular endothelial growth factor (hVEGF) and human interleukin-1 receptor antagonist (hIL-1Ra). Human islets were co-cultured with hBMSCs overexpressing hVEGF and hIL-1Ra.
View Article and Find Full Text PDFPLoS One
January 2014
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Centre, SunYat-sen University, Guangzhou, Guangdong Province, China.
Purpose: To investigate the protective effects of interleukin-1 receptor antagonist (IL-1ra) gene transfer in a rat model of corneal graft rejection.
Methods: We constructed a recombinant plasmid (pcDNA3.1-hIL-1ra) with high IL-1ra expression in eukaryotic cells.
Gene Ther
June 2013
Department of Orthopaedics and Rehabilitation, University of Florida, Gainesville, FL 32608-0137, USA.
With the long-term goal of developing a gene-based treatment for osteoarthritis (OA), we performed studies to evaluate the equine joint as a model for adeno-associated virus (AAV)-mediated gene transfer to large, weight-bearing human joints. A self-complementary AAV2 vector containing the coding regions for human interleukin-1-receptor antagonist (hIL-1Ra) or green fluorescent protein was packaged in AAV capsid serotypes 1, 2, 5, 8 and 9. Following infection of human and equine synovial fibroblasts in culture, we found that both were only receptive to transduction with AAV1, 2 and 5.
View Article and Find Full Text PDFCurr Eye Res
November 2012
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou, PR China.
Objective: To investigate the efficiency of the transfection of PEGFP-IL-1ra plasmid via cation polymer mediation (poly-ethylenimine, PEI) by injection into the corneal stroma.
Methods: Plasmid PEGFP-hIL-1ra recombinants were constructed and transferred into corneal endothelial cells via cation polymer mediation. Plasmid PEGFP-hIL-1ra recombinants and/or PEI-in-vivo were injected into the corneal stroma of Wistar rats.
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