摘要
【目的】可溶性表达结核分枝杆菌Ag85A蛋白,并评价其免疫原性。【方法】利用冷休克表达质粒和含有伴侣质粒的大肠杆菌对Ag85A蛋白进行可溶性原核表达,并进行纯化与鉴定,通过C57BL/6小鼠模型对Ag85A蛋白的免疫原性,包括诱导机体特异性体液免疫应答和细胞免疫应答水平进行分析。【结果】重组菌诱导后裂解上清中检测到可溶性Ag85A蛋白的表达,经过亲和层析纯化收获了纯度在90%以上的Ag85A蛋白,Western blot鉴定显示其具有较好的免疫反应性。Ag85A蛋白免疫小鼠后,血清中可以检测到高水平的Ig G抗体效价,其中Ig G2b水平要高于Ig G1。通过特异性多肽、蛋白刺激脾脏和腹股沟淋巴结细胞可分泌高水平的IFN-γ、TNF-α等Th1型细胞因子。【结论】实现了Ag85A蛋白的可溶性表达,免疫特性评价显示Ag85A蛋白可诱导机体产生强烈的特异性体液免疫应答及Th1型的细胞免疫应答,从而为其进一步免疫学功能的研究奠定了重要基础。
[Objective] The aim of this study was to express Mycobacterium tuberculosis Ag85 A protein and to evaluate its immunogenicity in mice. [Methods] The cold expressed system and chaperone competent cells BL21 were combined to express Mycobacterium tuberculosis Ag85 A protein, then the protein was purified with affinity chromatography and identified by Western Blot analysis. [Results] The immunogenicity of the purified Ag85 A protein was evaluated in C57BL/6 mice. Results show that high level of specific Ig G was elicited in the serum, and the splenocytes and lymph node cells of immunized mice could produce more Th1 cytokines, such as IFN-γ and TNF-α,after stimulated with specific antigen. [Conclusions] Ag85 A protein can induce strong specific humoral and Th1 type cellular immune responses, providing an important biological material for further research and application.
出处
《微生物学报》
CAS
CSCD
北大核心
2016年第5期804-813,共10页
Acta Microbiologica Sinica
基金
国家"973计划"(2012CB518805)
江苏省重点研发计划(BE2015343)
扬州市自然科学基金(YZ2014027)
国家自然科学基金(31201882)
江苏高校优势学科建设工程资助项目~~
作者简介
陈祥，chenxiang@yzu．edu．cn
通信作者。Tel：＋86．514．87971136；E-mail：焦新安，jiao@yzu．edu．cn；
