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

doi:10.11843/j.issn.0366-6964.2022.04.022

摘要/Abstract

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100 μg/100 μL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。

关键词: 热带螨, 变应原, 真核表达载体, DNA疫苗

Abstract: 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 100 μg/100 μL 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.

Key words: Blomia tropicalis, allergen, eukaryotic expression vector, DNA vaccine

中图分类号:

S852.746

罗应, 彭美琪, 肖正泮, 罗承慧, 王大勇, 裴业春. 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[J]. 畜牧兽医学报, 2022, 53(4): 1220-1230.

LUO Ying, PENG Meiqi, XIAO Zhengpan, LUO Chenghui, WANG Dayong, PEI Yechun. Expression of Blomia tropicalis Major Allergens Blo t 5 and Blo t 21 Induced the Production of IgG and Regulatory T Cells in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1220-1230.

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