VP1, a capsid protein of swine vesicular disease virus, was cloned from the SVDV HK/70 strain and inserted into retroviral vector pBABE puro, and expressed in PK15 cells by an retroviral expression system. The ability...VP1, a capsid protein of swine vesicular disease virus, was cloned from the SVDV HK/70 strain and inserted into retroviral vector pBABE puro, and expressed in PK15 cells by an retroviral expression system. The ability of the VP1 protein to induce an immune response was then evaluated in guinea pigs. Western blot and ELISA results indicated that the VP1 protein can be recognized by SVDV positive serum, Furthermore, anti-SVDV specific antibodies and lymphocyte proliferation were elicited and increased by VP1 protein after vaccination. These results encourage further work towards the development of a vaccine against SVDV infection.展开更多
目的:观察不同免疫途径和佐剂类型对柯萨奇病毒B组3型(Coxsackievirus group B type 3,CVB3)衣壳蛋白VP1免疫效果的影响。方法:将原核表达质粒pET-his/VP1转入E.coli BL21(DE3)pLysS中,用异丙基-1-硫代-β呋喃半乳糖苷(IPTG)诱导CVB3VP...目的:观察不同免疫途径和佐剂类型对柯萨奇病毒B组3型(Coxsackievirus group B type 3,CVB3)衣壳蛋白VP1免疫效果的影响。方法:将原核表达质粒pET-his/VP1转入E.coli BL21(DE3)pLysS中,用异丙基-1-硫代-β呋喃半乳糖苷(IPTG)诱导CVB3VP1蛋白的表达并进行纯化。首先采用不同的免疫途径(皮下,腹腔,肌肉)用VP1蛋白免疫小鼠,每组12只。然后另取小鼠分为PBS组和不同佐剂组(氢氧化铝、弗氏佐剂、Montanide ISA720),每组18只,采用肌肉注射途径免疫。每次每只小鼠注射50μg,共免疫3次,间隔3周。用ELISA和微量中和试验检测血清特异性IgG抗体和中和抗体。用CCK-8法检测淋巴细胞增殖活性和CTL杀伤活性。用致死量的CVB3攻击后,检测血中病毒的滴度并观察小鼠的存活状况。结果:在大肠杆菌中成功表达CVB3VP1蛋白。三种免疫途径比较,肌肉注射组血清中和抗体和特异性IgG抗体的水平明显高于其他组(P<0.01)。采用肌肉注射免疫时,弗氏佐剂组Montanide ISA 720佐剂组的体液免疫和细胞免疫应答的水平明显高于氢氧化铝组(P<0.05);但血中病毒的滴度低于氢氧化铝组(P<0.05)。弗氏佐剂组小鼠的生存率好于氢氧化铝组(P<0.05)。结论:采用肌肉注射途径,并联合弗氏佐剂或Montanide ISA 720佐剂可以使CVB3VP1免疫获得较好的免疫效果。展开更多
人肠道病毒71型（enterovims 7l，EV71）是婴幼儿手足口病（hand，foot and mouth disease，HFMD）的主要致病原之一。临床研究发现，EV71感染可导致多种临床表现，从轻微HFMD到致命性脑干脑炎并发肺水肿甚至死亡，但原因尚不清楚。EV7...人肠道病毒71型（enterovims 7l，EV71）是婴幼儿手足口病（hand，foot and mouth disease，HFMD）的主要致病原之一。临床研究发现，EV71感染可导致多种临床表现，从轻微HFMD到致命性脑干脑炎并发肺水肿甚至死亡，但原因尚不清楚。EV71衣壳蛋白VPl在病毒识别、结合、进入靶细胞及病毒颗粒装配过程中发挥核心作用，VPl变异是EV7l适应性和免疫原性的重要决定因素。现对VPl结构、功能及其相关的抗病毒疫苗和药物的研究进展进行综述，为进一步研发新的更有效的抗病毒疫苗和药物提供理论基础。展开更多
文摘VP1, a capsid protein of swine vesicular disease virus, was cloned from the SVDV HK/70 strain and inserted into retroviral vector pBABE puro, and expressed in PK15 cells by an retroviral expression system. The ability of the VP1 protein to induce an immune response was then evaluated in guinea pigs. Western blot and ELISA results indicated that the VP1 protein can be recognized by SVDV positive serum, Furthermore, anti-SVDV specific antibodies and lymphocyte proliferation were elicited and increased by VP1 protein after vaccination. These results encourage further work towards the development of a vaccine against SVDV infection.
文摘目的:观察不同免疫途径和佐剂类型对柯萨奇病毒B组3型(Coxsackievirus group B type 3,CVB3)衣壳蛋白VP1免疫效果的影响。方法:将原核表达质粒pET-his/VP1转入E.coli BL21(DE3)pLysS中,用异丙基-1-硫代-β呋喃半乳糖苷(IPTG)诱导CVB3VP1蛋白的表达并进行纯化。首先采用不同的免疫途径(皮下,腹腔,肌肉)用VP1蛋白免疫小鼠,每组12只。然后另取小鼠分为PBS组和不同佐剂组(氢氧化铝、弗氏佐剂、Montanide ISA720),每组18只,采用肌肉注射途径免疫。每次每只小鼠注射50μg,共免疫3次,间隔3周。用ELISA和微量中和试验检测血清特异性IgG抗体和中和抗体。用CCK-8法检测淋巴细胞增殖活性和CTL杀伤活性。用致死量的CVB3攻击后,检测血中病毒的滴度并观察小鼠的存活状况。结果:在大肠杆菌中成功表达CVB3VP1蛋白。三种免疫途径比较,肌肉注射组血清中和抗体和特异性IgG抗体的水平明显高于其他组(P<0.01)。采用肌肉注射免疫时,弗氏佐剂组Montanide ISA 720佐剂组的体液免疫和细胞免疫应答的水平明显高于氢氧化铝组(P<0.05);但血中病毒的滴度低于氢氧化铝组(P<0.05)。弗氏佐剂组小鼠的生存率好于氢氧化铝组(P<0.05)。结论:采用肌肉注射途径,并联合弗氏佐剂或Montanide ISA 720佐剂可以使CVB3VP1免疫获得较好的免疫效果。
文摘人肠道病毒71型（enterovims 7l，EV71）是婴幼儿手足口病（hand，foot and mouth disease，HFMD）的主要致病原之一。临床研究发现，EV71感染可导致多种临床表现，从轻微HFMD到致命性脑干脑炎并发肺水肿甚至死亡，但原因尚不清楚。EV71衣壳蛋白VPl在病毒识别、结合、进入靶细胞及病毒颗粒装配过程中发挥核心作用，VPl变异是EV7l适应性和免疫原性的重要决定因素。现对VPl结构、功能及其相关的抗病毒疫苗和药物的研究进展进行综述，为进一步研发新的更有效的抗病毒疫苗和药物提供理论基础。
