热带无爪螨主要变应原Blo t 5和Blo t 21融合表达质粒诱导小鼠产生IgG和调节性T细胞  

Expression of Blomia tropicalis Major Allergens Blo t 5 and Blo t 21 Induced the Production of IgG and Regulatory T Cells in Mice

作　　者：罗应 彭美琪 肖正泮 罗承慧 王大勇[1,2] 裴业春[1,2] LUO Ying;PENG Meiqi;XIAO Zhengpan;LUO Chenghui;WANG Dayong;PEI Yechun(School of Life Sciences, Hainan University, Haikou 570228, China;One Health Institute, Hainan University, Haikou 570228, China)

机构地区：[1]海南大学生命科学学院,海口570228 [2]海南大学全健康研究院,海口570228

出　　处：《畜牧兽医学报》2022年第4期1220-1230,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：海南省科技专项资助(ZDYF2022SHFZ059);海南省自然科学基金高层次人才项目(320RC467);国家自然科学基金(31860726,32160837)。

摘　　要：旨在开发防治热带螨过敏症的兽用核酸疫苗,本试验通过对热带无爪螨主要变应原Blo t 5和Blo t 21的基因序列进行密码子优化,构建了热带螨主要变应原融合基因的真核表达载体pVAX1-PADRE-Blo t 5-21-CpG。为验证该真核表达载体的免疫效果,采用皮下注射,以100μg/100μL量的pVAX1-PADRE-Blo t 5-21-CpG核酸免疫BALB/c小鼠(n=6),通过ELISA检测小鼠血清中特异性IgG、IgG1和IgG2a的水平变化;流式细胞术检测小鼠脾淋巴细胞中调节性T细胞的变化。结果显示,真核表达质粒pVAX1-PADRE-Blo t 5-21-CpG免疫小鼠后可诱导特异性IgG和IgG2a水平升高,且IgG2a/IgG1的比值升高,这暗示免疫后小鼠机体发生偏向Th1型的免疫反应。此外,该核酸疫苗可诱导小鼠产生较高水平的CD4+CD25+Foxp3+T细胞,这表明该真核表达质粒免疫小鼠后可能会诱发免疫抑制。To develop DNA vaccine against Blomia tropicalis allergy,we constructed eukaryotic expression vector pVAX1-PADRE-Blo t 5-21-CpG of major allergen fusion gene of Blomia tropicalis by codon optimization of Blo t 5 and Blo t 21 gene sequences.To verify the immune effect of the eukaryotic expression vector,BALB/c mice were immunized with 100μg/100μL of pVAX1-PADRE-Blo t 5-21-CpG by subcutaneous injection(n=6),and the serum levels of specific IgG,IgG1 and IgG2a were detected by ELISA.The changes of regulatory T cells in spleen lymphocytes of mice were detected by flow cytometry.The results showed that the eukaryotic expression plasmid pVAX1-PADRE-Blo t 5-21-CpG could induce the increase of specific IgG and IgG2a levels in mice,and the increase of IgG2a/IgG1 ratio suggested a bias towards a Th1 type immune response in mice after immunization.In addition,the plasmid pVAX1-PADRE-Blo t 5-21-CpG can induce higher levels of CD4+CD25+Foxp3+T cells in mice,suggesting that the eukaryotic expression plasmid may induce immunosuppression.

关 键 词：热带螨 变应原 真核表达载体 DNA疫苗 

分 类 号：S852.746[农业科学—基础兽医学]