马链球菌马亚种3种抗原蛋白对小鼠免疫的免疫原性分析

doi:10.11843/j.issn.0366-6964.2021.03.019

摘要/Abstract

摘要： 旨在对比研究马链球菌马亚种（Streptococcus equi subsp.equi，S.equi）3种不同抗原蛋白——分选酶A （sortase A，SrtA）、M蛋白（SeM）及IgG结合蛋白（EAG）的免疫原性及免疫保护效果，本研究以表达并纯化的SrtA、SeM及EAG 3种蛋白单独及联合免疫小鼠，免疫后检测了血清抗体效价、抗体亚型、攻毒保护率，以real-time PCR定量检测免疫相关分子，并对攻毒后的肝、脾、肺、肾荷菌量进行检测和病理组织学观察。结果显示：联合免疫组诱导产生的抗体效价及IgG、IgG1和IgG2b的抗体水平均高于各种蛋白单独免疫组，SrtA诱导的IgG2a抗体水平高于SeM和EAG免疫组；攻毒保护力、荷菌量及病理组织学观察结果表明，联合免疫组优于SeM和EAG免疫组；免疫相关分子的荧光定量PCR检测结果显示，SrtA诱导的MHCⅠ、TCR、TLR2、TLR3及TLR4的转录水平显著高于SeM、EAG免疫组和对照组，3种抗原联合免疫可显著提高MHCⅠ分子及TLR3分子的转录水平。综上表明，SrtA具有较强的免疫调节和诱导能力，3种蛋白联合免疫可诱导产生较高的免疫水平和良好的免疫保护效果。该结果为马腺疫亚单位疫苗的研究提供了理论参考和试验基础。

关键词: 马链球菌马亚种, 亚单位抗原蛋白, 免疫原性, 小鼠模型

Abstract: This study aims to evaluate and compare the immunogenicity and protective efficacy of virulence proteins, SrtA, SeM and EAG of Streptococcus equi subsp. equi (S. equi). These 3 recombinant proteins were expressed, purified, and then were used alone or in combination to immunize mice. The mice serum antibody titer, antibody subtype, and immune molecules were detected after immunization. Challenge protection rate, bacterial loading of liver, spleen, kidney, and lung were detected after challenge. In addition, histopathological changes of liver, spleen, kidney, and lung were observed. The ELISA results showed that IgG, IgG1 and IgG2b levels induced by the combined immunization group were higher than those immunized alone group and SrtA immunization group induced higher IgG2a levels than SeM and EAG group. The results of challenge protection rate, bacterial load and histopathological observation showed that the combined immunization group was superior to the single immunization group. The fluorescence quantitative PCR results of immune related molecules showed that the transcription levels of MHCⅠ, TCR, TLR2, TLR3 and TLR4 induced by SrtA were higher than those induced by SeM,EAG immunized group and control group. The MHC Ⅰ and TLR3 induced by combined-immunized group were higher than those of single-immunized group. In conclusion, the SrtA was a better immune stimulation and induction antigen and three proteins immunized in combination has better immunogenicity and provide efficacious immune protection. This data laid theory foundation and experiment basis for the subunit vaccine development of strangles.

Key words: Streptococcus equi, subunit antigen protein, immunogenicity, mice model

中图分类号:

S852.611

吕芬芬, 马晓慧, 汪丽, 马陈, 张宝江, 苏艳. 马链球菌马亚种3种抗原蛋白对小鼠免疫的免疫原性分析[J]. 畜牧兽医学报, 2021, 52(3): 752-762.

LÜ Fenfen, MA Xiaohui, WANG Li, MA Chen, ZHANG Baojiang, SU Yan. Immunogenicity Analysis of Three Different Antigen Proteins of Streptococcus equi in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 752-762.

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