重组猪IL-2对口蹄疫重组腺病毒免疫增强效果的研究  

Recombinant porcine interleukin- 2 enhances immune efficacy of candidate recombinant adenovirus vaccine against foot and mouth disease virus

作　　者：苏春霞[1] 段相国[2] 曹瑞兵[3] 郑洁[1] 黄菱[1] 郭琳[1] 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,银川750004 [2]宁夏医科大学基础医学院检验学院临床免疫学检验系,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,210095

出　　处：《免疫学杂志》2014年第2期110-113,共4页Immunological Journal

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

摘　　要：目的构建表达猪白细胞介素-2(poIL-2)的重组腺病毒,研究其作为佐剂对口蹄疫重组腺病毒免疫效果的影响。方法将poIL-2基因克隆到腺病毒的穿梭载体中,与腺病毒骨架载体共转化BJ5183感受态细胞,获得的重组病毒质粒通过脂质体转染HEK-293A细胞,获得表达poIL-2的重组腺病毒(rAd5poIL-2)。以rAd5poIL-2作为佐剂与表达PMDV VP基因的重组腺病毒(rAd5VP1)联合免疫小鼠,以单独免疫rAd5VP1的小鼠作为对照。通过测定淋巴细胞增殖指数、特异性抗体、IL-4和IFN-γ的水平来衡量rAd5poIL-2对rAd5VP1的免疫增强效果。结果经PCR鉴定,成功构建了rAd5poIL-2。MTT与抗体检测结果显示,rAd5poIL-2既能增强rAd5VP1诱导小鼠淋巴细胞增殖,又能促进rAd5VP1诱导小鼠特异性抗体的产生,细胞因子检测结果显示,rAd5poIL-2能促进rAd5VP1诱导Th1和Th2的分化。结论 rAd5poIL-2可望成为口蹄疫疫苗的候选佐剂。This study aimed to construct a recombinant adenovirus expressing porcine interleukin-2, and to study its immune adjuvant effects enhancing immunization of FMDV recombinant adenovirus. Porcine IL-2 gene was cloned into pAdenoVator-CMV5-IRES-GFP to construct pASPoIL-2. Then linearized pA5PolL-2 and pAdenoVatorAE1/E3 co-transformed into E. coli BJ5183 competent cells by electroporation. The homologous recombinant plasmids pAdSPoIL-2 was linearized and transfected into HEK-293A cells by Lipofectamine TM2000, and recombinant viruses were identified and designated rpAdSPoIL-2. Mice were inoculated intraperitoneal with recombinant virus rAdSpolL-2/rAd5VP1, rAdSVP1, and adenovirus as control. T lymphocyte proliferation index was tested by MTT, while levels of IgG, IL-4 and IFN-γ in mouse serum were quantitatively measured by ELISA. PCR results showed that rAd5poIL-2 was constructed successfully and MTT results showed that rAd5polL-2 has biological activity. Results of MTr and specific antibody detection showed rAd5poIL-2 promoted rAd5VP1 to induce not only T lymphocyte proliferation but also specific antibody secretion. In conclusion, rAdSpolL-2 could enhance immunization of rAd5VP1 by inducing differentiation of Thl and Th2 cells, and may be a prospective candidate for FMD vaccine adjuvant.

关 键 词：猪白细胞介素2 重组腺病毒 口蹄疫 细胞因子佐剂 

分 类 号：R392[医药卫生—免疫学]