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当前位置: 首页 > 文章 > 家蚕核型多角体病毒表面展示猪流行性腹泻病毒S1蛋白及其黏膜免疫原性研究 蚕业科学 2019 (2) 218-224
Position: Home > Articles > 家蚕核型多角体病毒表面展示猪流行性腹泻病毒S1蛋白及其黏膜免疫原性研究 Science of Sericulture 2019 (2) 218-224

家蚕核型多角体病毒表面展示猪流行性腹泻病毒S1蛋白及其黏膜免疫原性研究

作  者：
刘轶轩;严婷婷;徐萌;卢天翼;舒建洪;陈健
关键词：
家蚕核型多角体病毒;BmN细胞;猪流行性腹泻病毒;S1蛋白;黏膜免疫
摘  要：
猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)可以通过呼吸道感染在猪群中传播,提示呼吸道粘膜也许可以作为疫苗免疫接种的位点.PEDV通过表面的纤突蛋白S识别受体并侵入宿主细胞,该蛋白质S1区(1~735 aa)是PEDV主要的中和位点以及受体结合域,参与病毒吸附入侵宿主细胞.利用家蚕核型多角体病毒在BmN细胞中表达融合蛋白S1-eGFP并展示于病毒表面,Western blot鉴定融合蛋白分子质量约为150 kD.以重组杆状病毒为免疫原雾化免疫BALB/c小鼠,结果表明免疫16 d后小鼠血清中S1特异性IgG抗体效价可达1∶3 200,血清和肺气管润洗液中都能检测到高水平的S1特异性sIgA,说明引起了较强的黏膜免疫反应;脾淋巴细胞增殖结果进一步表明重组杆状病毒雾化免疫可以有效地诱导小鼠产生细胞免疫.研究结果初步表明,表面展示S1的重组杆状病毒可以作为PEDV黏膜免疫候选疫苗,具有进一步开发研究的价值.
作  者：
Liu Yixuan;Yan Tingting;Xu Meng;Lu Tianyi;Shu Jianhong;Chen Jian;College of Life Sciences and Medicine, Zhejiang Sci-Tech University;Zhejiang Provincial Key laboratory of Silkworm Bioreactor and Biomedicine;
单  位：
Liu Yixuan%Yan Tingting%Xu Meng%Lu Tianyi%Shu Jianhong%Chen Jian%College of Life Sciences and Medicine, Zhejiang Sci-Tech University%Zhejiang Provincial Key laboratory of Silkworm Bioreactor and Biomedicine
关键词：
Silkworm baculovirus;;BmN cell;;Porcine epidemic diarrhea virus;;S1 protein;;Mucosal immunity
摘  要：
Porcine epidemic diarrhea virus(PEDV) can be transmitted in pig farms through respiratory tract infection, suggesting that respiratory tract mucosa may serve as a site for vaccination. PEDV recognizes the receptor through the surface spike protein S and invades the host cell. S1 region(1-735 aa) of this protein contains the main neutralizing epitopes and receptor binding domain of PEDV, which is closely related to virus antigenicity and adsorption invasion. The fusion protein S1-eGFP was expressed in BmN cells by silkworm baculovirus and displayed on the surface of the virus, and Western blot analysis indicated that the molecular mass of the protein was about 150 kD. The recombinant virus as immunogen was used to vaccinate BALB/c mice by nebulization. Our data showed that the titers of S1 specific IgG in mouse serum got up to 1∶ 3 200 on the 16 th day after vaccination, and a relatively high level of S1 specific sIgA could be detected in both serum and alveolar lavage fluid from immunized mice, sugges-ting that some extent of mucosal immune response was elicited to PEDV. Further spleen lymphocyte proliferation showed that recombinant baculovirus nebulization immunization could effectively induce mice to produce cellular immunity. The results of this study preliminarily indicated that the recombinant baculovirus displaying S1 could be used as a candidate vaccine for mucosal immune against PEDV infection, which has the value for further development and research.

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