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Title

RECOMBINANT NONSTRUCTURAL 3 PROTEIN, RNS3, OF HEPATITIS C VIRUS ALONG WITH RECOMBINANT GP96 INDUCE IL-12, TNFα AND α5INTEGRIN EXPRESSION IN ANTIGEN PRESENTING CELLS

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Abstract

 Background: HEPATITIS C virus (HCV) infection is the main cause of chronic liver disease and to date there has been no vaccine development to prevent this infection. Among non-structural HCV proteins, NS3 protein is an excellent goal for a therapeutic vaccine, due to its large size and less variation in conserved regions. The immunogenic properties of heat shock proteins (HSPs) for instance GP96 have prompted investigations into their function as strong adjuvant to improve innate and adaptive immunity.Objectives: The aim of this study was to examine additive effects of recombinant GP96 (rGP96) fragments accompanied by rNS3 on expression levels of a5integrin and pro-inflammatory CYTOKINES, IL-12 and TNFa, in Antigen Presenting Cells (APCs).Materials and Methods: Recombinant viral proteins (rNS3 and rRGD-NS3), N-terminal and C-terminal fragments of GP96 were produced and purified from E. coli in order to treat the cells; mouse spleen Dendritic Cells (DCs) and THP-1 macrophages.Results: Our results showed that rNT-GP96 alone significantly increases the expression level of IL-12, TNFa and a5integrin in THP-1 macrophages and DCs, while IL-12 and TNFα expression levels were unaffected by either rNS3 or rRGD-NS3. Interestingly, the co-addition of these recombinant proteins with rNT-GP96 increased IL-12, TNFa and a5integrin expression. Pearson Correlation showed a direct association between α5integrin with IL-12 and TNF-a expression.Conclusions: we have highlighted the role of rNS3 plus rNT-GP96 mediated by α5integrin in producing IL-12 and TNFa. It can be suggested that rNT-GP96 could enhance immunity characteristic of rNS3 protein via production of pro-inflammatory CYTOKINES.

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