Moghaddam Pour M, Keivani H, Masoudi S H, Monavari S H, Najafi M
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.
Virology Department, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
J Med Life. 2015;8(Spec Iss 4):189-195.
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. 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. 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. 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.
两种结构抗原，即血凝素和神经氨酸酶，是开发流感候选疫苗的主要成分。重组疫苗通过简单方法生产，尽管有望诱导针对特定抗原的免疫反应，但其有效性仍有待进一步提高。一般来说，结核分枝杆菌的热休克蛋白70作为一种有效的佐剂，常用于改善抗原呈递细胞（APC）的功能，从而激发T淋巴细胞。本研究的目的是评估融合到HSP70 C末端的NA抗原作为候选疫苗在诱导针对疫苗抗原的强效、保护性免疫应答方面的疗效。通过聚合酶链反应增强NA基因，然后克隆到真核表达载体pFastBac HTA中。随后，在杆状病毒中表达与HSP70融合的重组NA蛋白。在SDS-PAGE电泳上研究表达的NA-HSP70融合蛋白的纯度。进行蛋白质印迹以研究NA-HSP70的表达。此外，使用免疫荧光测定法定性评估重组NA-HSP70蛋白在感染的Sf9细胞表面的生物学和抗原活性谱，方法是使用免疫兔抗血清。有趣的是，本研究结果表明，HSP蛋白具有刺激和增强强效体液免疫和细胞介导免疫反应的能力，并且与其他蛋白结合时起佐剂作用。因此，与HSP融合的重组蛋白为基于HSP的疫苗开发带来了希望。
