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乳酸菌NICE(乳链菌肽控制表达nisin controlled expression,NICE)系统可将治疗性蛋白或保护性抗原蛋白表达于细胞外或锚定于菌体的肽聚糖上,递呈抗原蛋白并激活粘膜免疫系统,刺激特异性s-IgA的产生使其作为粘膜免疫原递呈的活载体成为可能。本试验将构建的重组乳酸乳球菌PNZ8149/NZ3900-M/PRRS鼻腔免疫BALB/c小鼠,检测呼吸道粘膜中抗体s-IgA和血清中特异性抗体IgG含量,评价其动态变化情况,同时检测脾脏细胞因子IL-4和IL-10的活性。结果重组乳酸乳球菌PNZ8149/NZ3900-M/PRRS免疫小鼠后,在测定时间内重组菌试验组小鼠呼吸道粘膜中特异性s-IgA水平高于对照组,差异极显著((P<0.01);血清中特异性抗体IgG水显著高于对照组((P<0.01);脾脏细胞悬液中的IL-10和IL-4含量与对照组无差异性(p>0.05),结果表明重组菌PNZ8149/NZ3900-M/PRRS经粘膜途径免疫小鼠后能能刺激小鼠粘膜特异性抗体s-IgA和血清中特异性抗体IgG的分泌,且能避免粘膜免疫耐受的产生,为该重组疫苗的进一步应用奠定了理论基础。
Lactic acid bacteria NICE (NICE) system can express the therapeutic protein or protective antigen protein on the extracellular or peptidoglycan anchored to the cell body to present the antigen protein and activate the mucosal immune system to stimulate The production of specific s-IgA makes it possible to act as a live vector for mucosal immunogen presentation. BALB / c mice were immunized nasally with recombinant Lactococcus lactis PNZ8149 / NZ3900-M / PRRS to detect s-IgA and serum IgG in respiratory mucosa, and to evaluate the dynamic changes of spleen Activity of cytokines IL-4 and IL-10. Results The specific s-IgA level in the respiratory mucosa of the recombinant bacteria test group was significantly higher than that of the control group (P <0.01) after the recombinant Lactococcus lactis PNZ8149 / NZ3900-M / PRRS was immunized. (P <0.01). The levels of IL-10 and IL-4 in the spleen cell suspension were not significantly different from those in the control group (p> 0.05). The results showed that the recombinant bacteria Immunization of mice with PNZ8149 / NZ3900-M / PRRS by mucosal route can stimulate the secretion of mucosal-specific antibody s-IgA and serum-specific IgG in mice and avoid mucosal immune tolerance. The further application has laid a theoretical foundation.