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| Recombinant Expression of Antigen Epitope Genes of Human Cytomegalovirus and Immunogenicity Detection of the Fusion Protein |
| WANG Meng1, SUN Wen-hui1, JI Fang-ling1, PU Zhong-ji1, LI Yin-sheng2, BAO Yong-ming1 |
1. School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China;
2. Dalian Institute for Drug Control, Dalian 116029, China |
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Abstract Objective: To obtain the recomninant protein as HCMV subunit vaccine, the dominant antigen epitope gene of protein pp65 and gB of human cytomegalovirus (HCMV) were expressed in E. coli, and the humoral and cellular immune responses of the recombinant protein in Balb/c mice were detected. Methods: The coden genes of 490-508 aa in pp65 protein and 607-621 aa in gB protein were selected as gene segments and cloned into expression vector pET32a (+) after amplified by PCR. The protein expressed was analyzed by SDS-PAGE, after the recombinant plasmid transformed into E. coli BL21 (DE3) plys. Balb/c mice were immunized with the recombinant protein purified by nickel metal chelate affinity column. The humoral immune activity of recombinant proteins was detected by Western blotting and indirect ELISA. The cellular immune activity was detected by flow cytometry and sandwich ELISA. Results: The fusion protein about 22kDa was obtained. Western blotting test showed that the multiclonal antibody for HCMV has specificity and the titers reach to 1:102 400 detected by indirected ELISA. The number of CD4+ T cells and CD8+ T cells in peripheral blood and the quantities of IFN-γ, IL-2, IL-12 were significantly higher than control group (P<0.01). Conclusions:The recombinant protein with dominant antigen epitopes can induce significant humoral immunity and cellular immunity in mice, and is a potential vaccine for HCMV.
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Received: 23 January 2017
Published: 25 July 2017
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