鼻黏膜复合佐剂LTB-Rg1在小鼠体内对猪繁殖与呼吸综合征病毒灭活疫苗的增强作用及其生物安全性评价  

Enhancement effect of nasal mucosal adjuvant LTB-Rg1 on an inactivated porcine reproductive and respiratory syndrome virus vaccine in mice and its biological safety evaluation

作　　者：苏菲[1] 薛银 徐丽华[1] 李军星[1] 余斌[1] 袁秀芳[1] SU Fei;XUE Yin;XU Lihua;LI Junxing;YU Bin;YUAN Xiufang(Institute of Animal Husbandry and Veterinary Medicine,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;Zhejiang Center of Animal Disease Control,Hangzhou 310018,China)

机构地区：[1]浙江省农业科学院畜牧兽医研究所,杭州310021 [2]浙江省动物疫病预防控制中心,杭州310018

出　　处：《浙江大学学报（农业与生命科学版）》2022年第3期383-392,共10页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：浙江省基础公益研究计划项目(LGN21C180001);国家自然科学基金项目(31802235);浙江省重点研发计划项目(2021C02007-9);浙江省农业科学院青年人才培养项目(2017R22R08E03)。

摘　　要：为探究重组大肠埃希菌不耐热肠毒素B亚单位(B subunit of heat-labile enterotoxin,LTB)和人参皂苷Rg1联合滴鼻免疫小鼠的佐剂效果并评估其生物安全性,本研究分别将生理盐水,猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)灭活疫苗,PRRSV灭活疫苗混合LTB或Rg1或LTB-Rg1滴鼻免疫小鼠,共免疫3次。利用酶联免疫吸附测定检测小鼠血清和支气管肺泡黏液中的免疫球蛋白M(immunoglobulin M,IgM)、免疫球蛋白G(IgG)和免疫球蛋白A(IgA)抗体水平;采用实时荧光定量聚合酶链反应法检测小鼠脾脏和肺脏中细胞因子mRNA转录水平;定期监测小鼠的体质量变化和血清生化指标。结果显示:与单独PRRSV疫苗组相比,LTB-Rg1/PRRSV组能够在血清和呼吸道黏膜表面迅速产生高水平的IgM、IgG和IgA抗体,并有效延长抗体的作用时间;显著上调Th1、Th2型细胞因子的转录水平;对小鼠的体质量增长和肝肾功能均无明显影响。综上所述,LTB-Rg1是一种比较安全且极具潜力的鼻黏膜免疫佐剂,值得深入研究。The aim of this study was to investigate the effect of intranasal adjuvant B subunit of heat-labile enterotoxin(LTB)of the recombinant Escherichia coli in combination with ginsenoside Rg1 in mice and evaluate its biological safety profile.Mice were intranasally immunized with saline,inactivated vaccine of porcine reproductive and respiratory syndrome virus(PRRSV),inactivated vaccine of PRRSV admixed with LTB or Rg1 or LTB-Rg1 for three times,respectively.Immunoglobulin M(IgM),IgG and IgA antibody levels in serum and bronchoalveolar mucus were measured by enzyme-linked immunosorbent assay;mRNA expression levels of cytokines from spleen and lung were detected by real-time fluorescent quantitative polymerase chain reation;body mass and serum biochemical indexes of mice were monitored regularly.The results showed that LTB-Rg1 quickly increased the levels of PRRSV-specific IgM,IgG and IgA antibodies in serum and bronchoalveolar mucosa,prolonged antibody functioning time,and remarkably upregulated the expression levels of Th1-and Th2-type cytokines when compared with the vaccine alone group.LTB-Rg1 had no effects on body mass,hepatic or renal functions.Therefore,LTB-Rg1 is a safe and potential nasal mucosal immune adjuvant that is worthy of further study.

关 键 词：LTB-Rg1 猪繁殖与呼吸综合征病毒 鼻黏膜 复合佐剂 

分 类 号：S853.74[农业科学—临床兽医学]