诺如病毒病毒样颗粒疫苗免疫小鼠诱导的体液和黏膜免疫应答  

Humoral and mucosal immune responses induced by Norovirus-like particle vaccine in mice

作　　者：刘悦越[1] 赵荣荣[1] 赵一荣 张韵祺 杜加亮[3] LIU Yueyue;ZHAO Rongrong;ZHAO Yirong;ZHANG Yunqi;DU Jialiang(Division of Respiratory Virus Vaccines,National Institutes for Food and Drug Control,Beijing 102629,China;School of Mathematics and Statistics,Yunnan University;Division of Monoclonal Antibody Products,National Institutes for Food and Drug Control)

机构地区：[1]中国食品药品检定研究院呼吸道病毒疫苗室,北京102629 [2]云南大学数学与统计学院 [3]中国食品药品检定研究院单克隆抗体产品室

出　　处：《山西医科大学学报》2021年第12期1570-1575,共6页Journal of Shanxi Medical University

基　　金：国家科技重大专项“重大新药创制”项目(2018ZX09739002)。

摘　　要：目的观察GⅠ.1/GⅡ.4诺如病毒(Norovirus,NoV)病毒样颗粒疫苗免疫小鼠诱导的体液和黏膜免疫应答。方法7周龄BALB/c小鼠20只被随机分为对照组和疫苗组,每组10只。疫苗组每只小鼠腹腔注射0.1 ml疫苗,含GⅠ.1和GⅡ.4型衣壳蛋白VP1各10μg及0.08 mg氢氧化铝佐剂,对照组腹腔注射0.08 mg/0.1 ml氢氧化铝佐剂,第0,21天免疫。首次免疫后第7,14,21,28,35天,采集血清和粪便,测定血清GⅠ.1和GⅡ.4型IgG、IgG1、IgG2a、IgM、IgA、组织血型抗原(histoblood group antigen,HBGA)阻断抗体及粪便IgA。采用SPSS 23.0软件统计两组间和不同时间点各型抗体水平的差异。结果疫苗组小鼠第1剂免疫后第7天产生较高水平的NoV型特异性IgG、IgG1、IgG2a和IgM,IgA在第2剂免疫后第7天明显升高。疫苗组的各型抗体均高于对照组,两组间差异均有统计学意义(P<0.01)。疫苗组不同时间点的抗体,除GⅠ.1型IgG和IgG1外,其他差异均有统计学意义(P<0.01)。疫苗组首剂免疫后第28天和第35天的各型血清IgA抗体水平差异无统计学意义(P>0.05)。首剂免疫后第28天,粪便GⅠ.1型和GⅡ.4型IgA分别为160和80,血清GⅠ.1型和GⅡ.4型抗体阻断效价50分别为400和800。结论GⅠ.1/GⅡ.4诺如病毒疫苗可诱导小鼠产生体液和黏膜免疫应答,可能对NoV引起的疾病具有保护作用。Objective To investigate the humoral and mucosal immune responses induced by the candidate bivalent GⅠ.1/GⅡ.4 Norovirus-like particle(NoV VLP) vaccine in mice. Methods Twenty 7-week-old BALB/c mice were divided into vaccine group and control group, with 10 mice in each group. The mice in vaccine group were immunized intraperitoneally with 0.1 ml vaccine containing 10 μg GⅠ.1 and 10 μg GⅡ.4 VP1 proteins and 0.08 mg aluminum hydroxide adjuvant. The mice in control group were immunized with 0.08 mg adjuvant in 0.1 ml. The mice were immunized at day 0 and day 21. Serum and feces samples were collected at day 7, 14, 21, 28 and 35 after the first dose to detect serum GⅠ.1 and GⅡ.4 type-specific antibodies(IgG, IgG1, IgG2 a, IgM, IgA and HBGA blocking antibodies) and fecal IgA. The statistical analysis was performed using SPSS 23.0 software to evaluate the differences of humoral and mucosal responses between the two groups and among different time points with P<0.05 as test level of significance. Results Levels of NoV specific humoral and mucosal responses were increased, including IgG, IgG1, IgG2 a and IgM at day 7 after the first dose and IgA at day 7 after boosting in vaccine group when compared with control group. The antibody levels were higher in vaccine group than in control groups, and there was statistical significance in the antibody levels at different time points except GⅠ.1 type-specific IgG and IgG1. In addition, serum IgA in vaccine group reached to the peak at day 28 after the first dose and was comparable to that at day 35(P>0.05). The titers of fecal IgA and HBGA blocking antibodies reached to 160(GⅠ.1)/80(GⅡ.4) and 400(GⅠ.1)/800(GⅡ.4) at day 28 after the first dose, respectively. Conclusion The candidate bivalent GⅠ.1/GⅡ.4 NoV VLP vaccine induces strong humoral and mucosal immune responses in mice, which may provide a protective effect against diseases caused by NoV.

关 键 词：诺如病毒 疫苗 病毒样颗粒 体液免疫 黏膜免疫 抗体应答 

分 类 号：R392.7[医药卫生—免疫学]