猪流感病毒多表位抗原的小鼠黏膜免疫试验

Mucosal immunization of swine influenza virus multi-epitope antigens in mice

为探讨经黏膜途径免疫接种猪流感病毒(SIV)多肽疫苗的可行性,选择STV主要结构蛋白血凝素(HA)的T、B抗原表位,将多个表位片段串联,定向克隆到原核表达载体pET-30a(+)中,对串联表达的重组猪流感病毒蛋白SIV-ep1、SW-ep2、SIV-el2进行了小鼠免疫试验。重组蛋白以包涵体形式表达,经洗涤定量后,采用灌胃途径免疫6周龄KM小鼠,检测免疫小鼠抗体及细胞免疫水平。结果发现三免后各免疫组血清IgG抗体与PBS对照组差异极显著(P<0.01);黏膜分泌型IgA抗体,所有免疫蛋白组与PBS组差异均极显著(P<0.01);细胞免疫检测,所有免疫组小鼠外周血T淋巴细胞增殖明显;猪流感病毒血凝抗体检测,SIV-ep1免疫组、SIV-ep2免疫组、SIV-ep1与LT蛋白免疫组抗SIVH1N1亚型HI效价达1:1024,SIV-el2蛋白免疫组HI效价达1:512。结果表明,猪流感病毒重组多表位抗原灌胃免疫小鼠,能产生理想的黏膜免疫抗体和血清抗体。

To explore the feasibility of swine influenza virus （SIV） peptide vaccine by mucosal immunization, SIV-epl, SIV-ep2 and SIV- el2 were used as antigens to orally vaccinate into 6-week-old KM mice. Antibody and cellular immunity levels were detected in mice. The resuits showed that there were significant differences （ P〈0. 01 ） in serum IgG and secretory IgA levels between immunization and PBS groups. The detection of cell-mediated immunity indicated that, T lymphocyte proliferations of the immunized mice were significant in peripheral blood. The HI antibody titers to SIV H1N1 were up to 1 ： 1024 in the three groups, SIV-epl, SIV-ep2 and SlV-epl combined with heat-labile toxin （UF） of Escherichia coli, while 1 ： 512 in SIV-eL2 group. It suggested that, the ideal mucosal immunity and serum antibodies could be produced in mice by oral administration of SIV multi-epitope antigens.