摘要
目的探讨分子佐剂IL-17、IL-2对铜绿假单胞菌(Pa)外膜蛋白OprI免疫效果的影响。方法分别以IL-17、IL-2作为分子佐剂用Pa外膜蛋白OprI于第0、2、4周免疫雌性C57BL/6小鼠,同时设立单独的佐剂与蛋白对照组。末次免疫2周后,ELISA检测各组小鼠血清IgG、IgA以及IgG亚类水平;ELISA法检测小鼠脾细胞上清中Th1/Th17型细胞因子INF-γ、IL-2、IL-17以及Th2型细胞因子IL-4、IL-6水平;MTT法检测IL-17、IL-2分子佐剂对小鼠脾细胞增殖情况的影响,并以ConA作为阳性对照;末次免疫2周后Pa鼻内感染小鼠,感染后10 d计算各组小鼠死亡率并取小鼠肺组织检测菌体载量。结果 ELISA结果显示,OprI-IL-17、OprI-IL-2组小鼠血清中IgG、Ig A以及IgG亚类水平均显著高于各对照组(P<0.05),且其中各实验组IgG2a/IgG1的比值均大于1;IL-17、IL-2佐剂能够显著提高小鼠脾细胞上清中Th1/Th17型细胞因子INF-γ、IL-2、IL-17水平(P<0.05),而Th2型细胞因子IL-4、IL-6水平则无显著差异(P>0.05);此外,OprI-IL-17、OprI-IL-2组的淋巴细胞增殖最为显著(P<0.05);相较于各对照组,OprI-IL-17、OprI-IL-2组能够显著降低小鼠肺组织菌体载量以及小鼠死亡率(P<0.05),保护效果显著提高。结论 IL-17、IL-2佐剂均能有效增强Pa外膜蛋白OprI的体液、细胞免疫水平以及抗感染保护作用。
This study was performed to investigate the immue effect of IL-17 and IL-2 as moledeular adjuvants on the Pseudomonas aeruginosa （Pa） outer membrane protein OprI. Female C57BL/6 mice were immunized by Pa outer membrane protein OprI with IL-17 or IL-2 as moledcular adjuvants in 0, 2, 4 weeks, while adjuvants or proteins alone served as controls. Then the levels of serum IgG, IgA and IgG subclasses were detected by ELISA at week 2 after the last immunization, and the levels of Th1/Th17 type cytokines （INF-γ IL-2, IL-17） and Th2 type cytokines （IL-4, IL-6） in the cell supernatant were detected by ELISA. Also, the effects of IL-17 and IL-2 adjuvants on the proliferation of mouse splenocytes were detected by MTT assay, with ConA as a positive control. The mice were intranasally infected by Pa at week 2 after the last immunization, then the mortality of mice in each group was calculated and the bacterial load of lung tissue was detected by quantitative experiment 10 days after challenge. ELISA results showed that the levels of IgG, IgA and IgG subclasses in serum of OprI-IL-17 or OprI-IL-2 groups were significantly higher than those in control group （P〈0.05）, and the ratio of IgG2a/IgG1 in each experimental group was greater than 1. IL-17 and IL-2 adjuvants could significantly increase the levels of Thl/ Thl7 cytokines INF-γ, IL-2 and IL-17 in the spleen cell supernatant of mice （P〈0.05）, but not change the levels of Th2 cytokines IL-4 and IL-6 （P〉0.05）. Moreover, OprI-IL-17 and OprI-IL-2 could significantly promote lymphocyte proliferation （P〈0.05）, which also could significantly reduce the bacterial load of lung tissue and mortality of mice （P〈0.05）, eompared to the control group. Those data suggest that IL-17 and IL-2 adjuvants can effectively enhance the effect of humoral and cellular immunity and protective immunity induced by Pa outer membrane protein OprI.
作者
卿蕊
向晴可
刘众齐
肖非
阳帆
QING Rui;XIANG Qingke;LIU Zhongqi;XIAO Fei;YANG Fan(Department of Pathogenic Biology, Laboratory Medicine College, Shaoyang University, Shaoyang 422000, China;Department of Basic Medicine, Xiangnan University, Chenzhou 423000, China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2018年第7期581-586,共6页
Immunological Journal
基金
湖南省教育厅科学研究项目(16C1438)
