补益类复方中药调控流感病毒样颗粒免疫小鼠外周血白细胞介素17 A的表达水平

摘要

Objective To explore the regulating effects of tonic herbal compound on the IL-17A expression induced by influenza virus-like particles in peripheral blood. Methods 42 BALB/C mice were divided into 7 groups(n= 6 for each group),involving single VLPs group,VLPs combined Yiqi recipe, VLPs combined Buxue recipe,VLPs combined Ziyin recipe,and VLPs combined Zhuangyang recipe group, VLPs combined cholera toxin group was considered as positive control,and PBS only group was considered as blank control. The mice were immunized with 10μg VLPs each time through intranasal administration in day 0 and 14,and herbs were intragastric administration with 200 mg/Kg from first day to day 13. The blood was collected to make serum and PBMCs at pre-immunization and day 13 and day 28 after immunization. The IL-17A level in serum was determined by ELISA,and spot forming cells( SFCs) of IL-17A were determined using Elispot assay. Results (1)No matter in the early stage or late stage,influenza VLPs could induce IL-17A expression in serum, especially, Yiqi recipe could increase IL-17A level compared to single VLPs group(P<0. 01). (2)Elispot technology can effectively detect the secretion of IL-17A positive cells in PBMCs. The SFCs of IL-17A was much higher in the Yiqi recipe and Ziyin recipe (P<0. 01),especially at day 28 after immunization. Conclusion Yiqi recipe and Ziyin recipe can effectively increase IL-17A expression induced by influenza VLPs,which could be considered as new type adjuvants for influenza vaccine.%目的：探讨补益类复方中药对流感病毒样颗粒( virus-like particles,VLPs)免疫小鼠外周血白细胞介素17A(interleukin 17A,IL-17A)表达的影响。方法将42只BALB/C小鼠随机分为VLPs组、VLPs+益气方组、VLPs+补血方组、VLPs+滋阴方组、VLPs+壮阳方组、VLPs+霍乱毒素组和PBS组共7组,每组6只,分别于0和14天通过鼻腔使用VLPs(10μg/次)进行免疫,联合中药组在第1至第13天连续灌服中药。中药佐剂按生药量200 mg/kg剂量接种。在免疫第0天、13天及28天采集外周血并分离血浆和单个核细胞(peripheral blood mononuclear cells,PBMCs),利用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测各组外周血浆IL-17A浓度,利用酶联免疫斑点法(enzyme-linked immunospot assay,Elispot)检测PBMC中分泌IL-17A的细胞数,即斑点形成集落数(spot forming cells,SFCs)。结果(1)无论是在免疫应答早期还是后期,VLPs能有效诱导外周血浆中IL-17A的分泌,而联合益气方组血浆中IL-17A水平进一步提高(P<0.05)；(2) Elispot技术能有效检测PBMCs中分泌IL-17A的阳性细胞。与单纯VLPs免疫组相比,益气方和滋阴方均能显著增加分泌IL-17A的细胞数(P<0.01),在免疫后28天尤为明显。结论益气方、滋阴方可以显著增强流感VLPs诱导的IL-17A表达水平,益气联合滋阴方有望成为新型流感疫苗佐剂。