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共表达口蹄疫病毒VP1-2A基因与猪IL-2重组腺病毒的构建及其免疫效果研究 被引量：3

Construction of Recombinant Adenovirus Co-expressing VP1-2A of Foot and Mouth Disease Virus and Porcine IL-2 and Its Immune Effects
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摘要 构建共表达口蹄疫病毒(FMDV)VP1-2A基因和猪白细胞介素2(IL-2)基因的重组腺病毒,并研究重组病毒对小鼠特异性免疫的影响。将FMDV VP1-2A基因和猪IL-2基因先后克隆到腺病毒的穿梭载体pAdenoVator-CMV5-IRES-GFP中,在受体菌中与腺病毒骨架载体pAdenoVatorΔE1/E3同源重组。重组腺病毒质粒转染HEK-293A细胞,得到重组腺病毒rAd5VP1-2A-Po IL-2。MTT法检测重组IL-2的生物活性。用rAd5VP1-2A-poIL-2免疫接种小鼠,同时设免疫单独表达FMDV VP1基因的重组腺病毒rAd5VP1和空腺病毒的对照组。通过检测淋巴细胞增殖功能,特异性抗体分泌,IL-4和IFN-γ的表达水平衡量rAd5VP1-2A-poIL-2对小鼠特异性免疫应答的影响。结果显示,成功获得了rAd5VP1-2A-poIL-2,MTT法检测重组IL-2具有生物学活性。小鼠免疫试验结果显示,与rAd5VP1免疫组相比,rAd5VP1-2ApoIL-2免疫既能增强重组病毒诱导小鼠淋巴细胞增殖的能力,又能提高重组病毒诱导小鼠特异性抗体的水平。细胞因子检测结果显示,串联IL-2能促进重组病毒诱导Th1和Th2的分化。研究结果说明,rAd5VP1-2A-poIL-2可望成为口蹄疫的候选疫苗。 To construct the recombinant adenovirus expressing VP1-2A and porcine IL-2 （poIL-2）, and study the specifici mmunity of recombinant adenovirus in mice,FMDV VP1-2A gene and poIL-2 gene were cloned into pAdenoVator-CMVS-IRES-GFP, and the recombinant plasmid was linearized and co-trans- formed into E. coli BJ5183 competent cells with pAdeno VatorAE1/E3 by electroporation. The homolo- gous recombinant plasmid pAd5VP1-2A-poIL-2 was selected and then transfected into HEK-293A cells, recombinant viruses was selected and named rAd5VP1-2A-poIL-2. The biological action of IL-2 was detec- ted by lymphocyte proliferation test. Mice were inoculated intraperitoneally with recombinant viruse rAd5VP1-2A-poIL-2 three times at 2 weeks intervals, and with rAd5VP1 and adenovirus vector as control. T lymphocyte proliferation test, levels of IgG,IL-4,IFN-7 were detected to evaluate humoral and cellular immune responses of recombinant viruses. RT-PCR results indicated that VP1 gene and poIL-2 gene were expressed in HEK-293A cells and MTT results showed that rAd5VP1-2A-poIL-2 has biological activity. After mice were immunized with recombinant adenovirus, rAd5VP1-2A-poIL-2 not only induced T lym- phocyte proliferation but also induced antibody secretion. MTT results revealed that rAd5VP1-2A-poIL-2 induced stronger cell immune responses than rAd5VP1. ELISA results indicated that rAd5VP1-2A-poIL-2 can improve FMDV specific antibody levels. Cytokine detection results showed that rAd5VP1-2A-poIL-2 not only promote the recombinant virus-induced Thl differentiation, but also promote the recombinant vi- rus-induced Th2 differentiation. Results revealed that rAd5VP1-2A-poIL-2 is expected to become a candi- date for FMD vaccine.
作者 苏春霞 段相国 郑洁 林源 郭琳 陈溥言
机构地区 宁夏医科大学基础医学院病原生物学与免疫学系 宁夏医科大学检验学院临床免疫学检验系 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
出处 《动物医学进展》 CSCD 北大核心 2014年第2期1-5,共5页 Progress In Veterinary Medicine
基金 国家自然科学基金项目(31001064) 国家自然科学基金项目(81260678) 国家自然科学基金项目(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)
关键词 口蹄疫病毒 重组腺病毒 VP1-2A基因 白细胞介素2 Foot-and-mouth disease virus recombinant Adenovirus VP1-2A gene interleukin-2
分类号 S852.659.6 [农业科学—基础兽医学]
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参考文献16

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动物医学进展
2014年 第2期

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