CVB3VP1蛋白和重组腺病毒单独及联合应用的免疫效果  

IMMUNOLOGICAL EFFECTS OF RECOMBINANT ADENOVIRUS VACCINE AD /VP1 COMBINED WITH PROTEIN VP1 OF COXSACKIEVIRUS B3 IN MICE

作　　者：田新利[1] 李伟[1] 李剑[1] 温婵[1] 李嘉[1] 闫立景[1] 谢立新[1] 王永祥[1] 

机构地区：[1]河北省邢台医学高等专科学校,邢台054000

出　　处：《现代预防医学》2010年第9期1721-1724,共4页Modern Preventive Medicine

基　　金：河北省科学技术研究与发展计划(08276101D-93)

摘　　要：[目的]观察柯萨奇病毒B3(coxsackievirus group B type3,CVB3)VP1蛋白和重组腺病毒Ad/vp1单独及联合应用的免疫效果。[方法]大量扩增重组腺病毒Ad/vp1,制备并纯化CVB3/VP1蛋白,肌肉注射免疫BALB/c小鼠。小鼠随机分为4组:腺病毒组:肌肉注射Ad/vp1,共2次;蛋白组:注射VP1蛋白,共3次;腺病毒/蛋白组:第1次注射Ad/vp1,第2、3次注射VP1蛋白;PBS对照组。各次注射间隔时间为3周。腺病毒每次接种4×107pfu/只,VP1蛋白每次接种100μg/只。用中和试验法和ELISA法分别检测每次免疫后血清中和抗体和IgG抗体滴度;CCK-8法检测淋巴细胞增殖活性和CTL杀伤活性;用致死量病毒感染后检测血中病毒滴度,并观察小鼠的生存情况。[结果]中和抗体滴度及血清特异性IgG抗体水平随免疫次数增加而提高,末次免疫后,蛋白组(71.26±1.31,33779.40±1.46)、腺病毒/蛋白组的(56.56±1.44,22286.09±1.36)中和抗体及IgG抗体滴度明显高于腺病毒组(16.82±1.37,800±1.63)(P﹤0.05);蛋白组、腺病毒/蛋白组、腺病毒组的CTL杀伤活性明显高于PBS组(P﹤0.05);用致死量CVB3感染后,蛋白组、腺病毒/蛋白组的血中病毒滴度低于其他组,而生存率高于其他组(P﹤0.05)。[结论]蛋白组、腺病毒/蛋白组免疫小鼠的效果明显优于腺病毒组和PBS组。[Objective] To observe the immunological effects of coxsackievirus group B3 （CVB3） VP1 protein alone or combined with recombinant adenovirus vector vaccine Ad / vp1 on CVB3 infection in mice. [Methods] Amplified plenty of Ad / vp1. Prepared and purified VP1 protein. Male BALB / c mice of 4-6 week old were randomly divided into four groups： Ad / vp1 group was inoculated Ad / vp1 for two times; protein group was injected VP1 protein for three times; Ad / vp1-protein group received Ad / vp1 for the first time and VP1 protein for the second and third times; control group was injected PBS. Mice were intramuscularly immunized into thigh at the intervals of three weeks with 100μg dose of VP1 protein or 4×107 pfu dose of Ad / vp1. The titers of sera CVB3 VP1 neutralizing antibody and IgG antibody were detected by trace neutralization assay and ELISA. Three weeks after the last immunization, the spleen lymphocytes proliferation and specific CTL cytotoxic activity were tested by CCK-8. Mice in each group were challenged with lethal dose of CVB3, the titers of sera virus were titrated and the protection efficacy against CVB3 was observed. [Results] Titers of CVB3-specific neutralizing antibody and IgG antibody in sera of mice in every experimental groups were growing along with the immune times （P﹤0.05）. The titers of neutralizing antibody and IgG antibody in protein immunized group （71.26 ± 1.31, 33 779.40 ± 1.46） and Ad / vp1-protein group （56.56 ± 1.44, 22 286.09 ± 1.36） were higher titers than those of mice immunized with Ad / vp1 （16.82 ± 1.37, 800 ± 1.63） （P﹤0.05） alone after the last immunization. The CTL level in the protein group, Ad / vp1-protein group or Ad / vp1 group were higher than PBS group （P﹤0.05）.After challenged with lethal dose of CVB3, the titers of blood virus in VP1 protein or Ad / vp1-protein immunized group were significantly lower than the other groups （P﹤0.05）. Survival rates of the mice in VP1 protein or Ad / vp1-protein immunized group were sign

关 键 词：CVB3 VP1基因 重组腺病毒疫苗 蛋白疫苗 小鼠 

分 类 号：R186[医药卫生—流行病学]