重组免疫融合肽Tα1-BP5的表达及其免疫佐剂特性研究  

Fusion expression of Thymopentin α1 (Tα1) and Bursopentin (BP5) immunopeptide and its adjuvant effect on avian influenza inactivated vaccine

作　　者：王臣[1] 张巫凡 杨静[1] 郭香玲[1] 李德元[2] 陈溥言[2] 

机构地区：[1]河南科技大学兽医肿瘤免疫学重点实验室,河南洛阳471003 [2]南京农业大学农业部动物细菌学重点实验室,江苏南京210095

出　　处：《中国预防兽医学报》2016年第4期326-330,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学青年基金(31101792);河南省高校青年骨干教师项目(2012GGJS-077)

摘　　要：胸腺素α1(Tα1)和法氏囊活性五肽(BP5)均具有重要的免疫学功能,但二者在蛋白水平上的联合应用尚未见报道。为探究重组表达融合肽Tα1-BP5(r Tα1-BP5)是否具有免疫佐剂特性,本实验设计合成融合肽Tα1-BP5基因,将其克隆至p ET-32a中进行原核表达,并采用MTT法检测其体外活性。同时以rTα1-BP5联合H9N2型禽流感病毒(AIV)灭活疫苗免疫小鼠,检测免疫后小鼠的HI抗体、IgG抗体亚型和细胞因子(IL-4和IFN-γ)的分泌水平,并通过攻毒试验评价其对小鼠的免疫保护作用。结果显示,rTα1-BP5在大肠杆菌中获得可溶性表达;rTα1-BP5能够增强机体免疫后HI抗体、平衡IgG1和IgG2a抗体、提高Th2型(IFN-γ)和Th1(IL-4)型细胞因子的分泌水平,表明rTα1-BP5能够同时增强机体体液和细胞免疫应答。动物免疫保护试验结果显示rTα1-BP5有助于鼠肺脏中H9N2型AIV的清除。以上结果提示,rTα1-BP5具有良好的免疫佐剂的潜能。本研究为新型疫苗佐剂的研究开发奠定了基础。Thymosin α1（Tα1） and Bursopentin（BP5） are both associated with immune regulation. To investigate the effect of fusion peptide of Tα1 and BP5（r Tα1-BP5） on enhancement of immune response, the gene of Tα1-BP5 was synthesized according to the preferential codons of E.coli, and cloned into p ET-32 a for expression in E.coli. Then, the mice were immunized with r Tα1-BP5 combined with inactivated H9N2 AIV vaccine. The titers of HI antibody, antibodies（Ig G1 and Ig G2a） sub-types,and levels of Th1-type cytokines（IFN-γ） and Th2-type cytokines（IL-4） were assessed subsequently; Meanwhile, the immunization protective effects were estimated by virus challenge experiment. The results showed that r Tα1-BP5 was expressed in E.coli, and it enhanced HI and Ig G sub-type antibodies titers and promoted the secretion of Th1- and Th2-type cytokines, which suggested that r Tα1-BP5 was able to enhance both the humoral and cellular immune responses. These data demonstrated that rTα1-BP5 had immune adjuvant potential to be further developed as a new vaccine adjuvant.

关 键 词：胸腺素Tα1 法氏囊活性五肽BP5 融合表达 免疫佐剂 

分 类 号：S852.4[农业科学—基础兽医学]