仙茅多糖作为流感疫苗佐剂对小鼠巨噬细胞的免疫增强作用

Immunological enhancement effects of curculigo orchioides polysaccharide as adjuvant of influenza vaccine on mouse macrophage

目的:从体内外2个方面探讨仙茅多糖作为流感疫苗佐剂对巨噬细胞的抗原提呈能力和释放活性因子的影响。方法:在体外实验中,以小鼠巨噬细胞株RAW264.7为研究对象,用流式细胞仪检测不同浓度的仙茅多糖对细胞表面CD80、CD86和MHC-Ⅱ表达量的影响,用ELISA法检测细胞培养上清中TNF-α、IL-1、IL-6的水平;在体内实验中,以昆明种小鼠作为研究对象,仙茅多糖与流感疫苗同时注射对小鼠进行免疫,免疫3d后提取各组小鼠腹腔巨噬细胞,对其免疫效果进行评价。结果:在体外实验中,不同浓度的仙茅多糖刺激RAW264.7细胞后,细胞表面CD80、CD86和MHC-Ⅱ都有不同程度表达上调,同时使培养液中TNF-α、IL-1和IL-6含量上升,与阴性对照组比较,差异显著(P<0.05,P<0.01);在体内实验中,不同浓度的仙茅多糖可使腹腔巨噬细胞表面CD80、CD86和MHC-Ⅱ都有不同程度表达上调(P<0.05,P<0.01)。结论:仙茅多糖在体内外可以促进小鼠巨噬细胞的抗原提呈作用,能增强流感疫苗的免疫效应,有望成为具有潜在应用价值的流感疫苗佐剂。

Objective： To discuss the effects of curculigo orchioides polysaccharide （COP） as adjuvant of influenza vaccine on antigen presenting ability and releasing activity factors of macrophage, from the two aspects both in vivo and in vitro. Methods： In vitro experiment, the mouse macrophage cell line RAW264.7 was studied. The effects of different doses curculigo orchioides polysaccharide on the expression levels of CD80, CD86 and MHC-Ⅱ were detected by FCM. The levels of TNF-a, IL-1 and IL-6 in cell culture supernatants were detected by ELISA. In vivo experiment, Kunming mice were studied, both curculigo orchioides polysaccharide and influenza vaccination were simultaneously injected to them. Three days after immunization, peritoneal macrophages in all mice were extracted to evaluate the immune effects. Results： In vivo experiment, after different doses curculigo orchioides polysaccharide stimulating RAW264.7 cells, the expression levels of CD80, CD86 and MHC-Ⅱ were all up-regulated in different degrees, and the contents of TNF-α, IL-1 and IL-6 in cell culture supernatants were all increased, and there were all significant differences compared with the control group （P〈0.05, P〈0.01）. In vitro experiment, different doses curculigo orchioides polysaccharide could make the expression levels of CD80, CD86 and MHC-Ⅱ on the surface of the peritoneal macrophages all up-regulate in different degrees （P〈0.05, P〈0.01）. Conclusion： Curculigo orchioides polysaccharide both in vivo and in vitro, can promote the antigen presentation macrophages in mice and can enhance the immune effect of influenza vaccine. It is expected to be an adjuvant for influenza vaccine with potential application value,