Construction of recombinant fusion protein of influenza, a virus neuraminidase and heat shock protein 70 gene: expression in baculovirus and bioactivity.
Background:Two structural antigens, hemagglutinin and neuraminidase, are a major component for the development of influenza vaccine candidates. Recombinant vaccines are produced by a simple method, although expected to induce an immune response to a specific antigen, remaining to be further improved for their high effectiveness. In general, heat shock protein 70 of Mycobacterium tuberculosis, as a potent adjuvant, is commonly used to improve antigen-presenting cell (APC) function and thereby elicit T lymphocytes.Objective:The purpose of this research was to evaluate the efficacy of the NA antigen fused to the C-terminus of HSP70, as a vaccine candidate, in the induction of potent, protective immune answers specific to the vaccine antigen.Material and Method:The NA gene was strengthened via a polymerase chain reaction and then cloned to a eukaryotic expressing vector pFastBac HTA. Subsequently, a recombinant NA protein fusing to HSP70 was expressed in Baculovirus. The purity of the expressed NA-HSP70 fusion protein was investigated on the SDS-PAGE electrophoresis. Western blot was carried out to investigate the expression of NA-HSP70. Additionally, an immunofluorescence assay was used qualitatively to assess the biological and antigenicity activity profiles of the protein of recombinant, NA-HSP70, on the infected Sf9 cell surface by using immunized rabbit antiserum.Result and conclusion:Interestingly, the findings in the present studies suggested that HSP proteins have the ability to both stimulate and increase potent humoral- and cell-mediated immune responses, and play an adjuvant role when combined with other proteins. Therefore, a recombinant protein fusing to HSP raised hope regarding the development of an HSP-based vaccine.
Virology Department, School of Medicine, Iran University of Medical Sciences, Tehran, Iran ; Research and Development Viral Vaccine Department, Razi Vaccine & Serum Research Institute, Karaj, Iran.
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Moghaddam Pour
Virology Department, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
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Keivani
Poultry Viral Vaccines Research Department, Razi Vaccine & Serum Research Institute, Karaj, Iran.
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Masoudi
Virology Department, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
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Monavari
Cellular and Molecular Research Center, Biochemistry Department, Iran University of Medical Sciences, Tehran, Iran.
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Najafi
cloning; heat shock protein (HSP); influenza; neuraminidase; reverse genetics