重组白介素-2增强口蹄疫病毒多表位疫苗免疫效果研究  

Enhancement of Immune Effect of Foot-and-Mouth Disease Multi-epitope Vaccineby Recombinant IL-2

作　　者：苏春霞[1] 段相国[2] 陈溥言[3] 

机构地区：[1]宁夏医科大学基础医学院病原生物学与免疫学系,宁夏银川750004 [2]宁夏医科大学检验学院临床免疫学检验系,宁夏银川750004 [3]南京农业大学农业部动物疫病诊断与免疫重点开放实验室,江苏南京210095

出　　处：《动物医学进展》2014年第12期1-4,共4页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31001064;81260678;31360600);宁夏自然科学基金项目(NZ13060);宁夏教育厅高等学校科学研究项目(NGY2012065)

摘　　要：通过研究重组白细胞介素-2(IL-2)对口蹄疫病毒(FMDV)多表位疫苗免疫效果的增强作用,为IL-2作为FMDV表位疫苗佐剂应用提供实验依据。6周龄雌性Balb/c小鼠,分为4组,第1组联合免疫表达猪IL-2的重组腺病毒(rAd5poIL-2)和串联表达FMDV多表位基因的重组腺病毒(rAd5EGS),第2、3和4组分别注射rAd5EGS、灭活疫苗和PBS,间隔2周免疫1次,共免疫3次。首免后每周采血分离小鼠血清,通过ELISA检测血清中特异性IgG;首免后6周检测血清中抗体亚型IgG1、IgG2a及细胞因子IL-4和IFN-γ的表达水平,同时分离脾细胞,MTT法检测淋巴细胞增殖指数。结果显示,联合免疫rAd5poIL-2和rAd5EGS既能增强rAd5EGS诱导小鼠特异性IgG、IgG1和IgG2a的分泌,又能增强其诱导淋巴细胞增殖能力,且免疫效果强于灭活疫苗;细胞因子检测结果显示,联合免疫rAd5poIL-2后能增强rAd5EGS诱导小鼠IL-4和IFN-γ的分泌,且其诱导IL-4和IFN-γ分泌能力也强于灭活疫苗。说明重组IL-2能有效增强FMDV多表位疫苗诱导抗体分泌能力和细胞免疫应答,rAd5poIL-2有望作为FMDV表位疫苗的候选佐剂。To investigate the recombinant interleukin-2 （IL-2）enhancing the immune effect of foot-and-mouse disease（FMD）multi-epitope vaccine in mice,to provide the experimental evidence for studying ap-plication of IL-2 as an adjuvant for FMD epitope vaccine,6 weeks old female Balb/c mice were divided into 4 groups,mice of group 1 were combined immunized with recombinant adenovirus expressing porcine IL-2 （rAd5poIL-2）and recombinant adenovirus expressing FMDV multi-epitopes （rAd5EGS）,mice of group2,3 and 4 were injected with rAd5EGS,inactivated vaccine and PBS respectively;injection once every two week intervals for 3 times.The sera from mice were collected every week after immunization and the specific IgG was detected by ELISA;the secretion levels of IgG1,IgG2a,IL-4 and IFN-γ in serum were measured by ELISA after 6 weeks;Spleen cells were isolated after 6 weeks and lymphocyte proliferation index was measured by MTT assay.The results of ELISA and MTT showed that combined immunization with rAd5poIL-2 and rAd5EGS not only significantly enhanced rAd5EGS inducing the secretion of specific IgG, IgG1 and IgG2a,but also boosted the mouse lymphocyte proliferation,and which even induced higher im-mune response than inactivated vaccine.Cytokine detection results showed that combined immunized with rAd5poIL-2 could enhance the secretion of IL-4 and IFN-γ compared with rAd5EGS only,and which also induced higher levels of cytokines than inactivated vaccine.The results indicated that recombinant IL-2 could effectively enhance the FMD multi-epitope vaccine inducing specific antibody secretion and cellular immune responses.rAd5poIL-2 as a potent candidate adjuvant for FMD epitope vaccine is expected to put into practice.

关 键 词：口蹄疫病毒 多表位疫苗 佐剂 白细胞介素-2 

分 类 号：Q81[生物学—生物工程]