藏猪白细胞介素-12基因表达对猪圆环病毒2 疫苗的免疫增强效应
Elevation of immunity to PCV2 vaccine by recombinant Tibetan pig interleukin-12 gene in chitosan nanoparticles
作者：宋婷玉(四川大学生命科学学院生物资源与生态环境教育部重点实验室, 动物疾病防控和食品安全四川省重点实验室)；肖永乐(四川大学生命科学学院生物资源与生态环境教育部重点实验室, 动物疾病防控和食品安全四川省重点实验室)；曾光志(四川省华派生物制药有限公司)；万小平(四川大学生命科学学院生物资源与生态环境教育部重点实验室, 动物疾病防控和食品安全四川省重点实验室)；陈祎(四川大学生命科学学院生物资源与生态环境教育部重点实验室, 动物疾病防控和食品安全四川省重点实验室)；李金海(四川省华派生物制药有限公司)；王泽洲(四川省动物疫病预防控制中心)；方鹏飞(四川省华派生物制药有限公司)；高荣(四川大学生命科学学院生物资源与生态环境教育部重点实验室, 动物疾病防控和食品安全四川省重点实验室)
Author：SONG Ting-Yu(Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Key Laboratory for Animal Disease Prevention and Food Safety of Sichuan Province, College of Life Sciences, Sichuan University)；XIAO Yong-Le(Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Key Laboratory for Animal Disease Prevention and Food Safety of Sichuan Province, College of Life Sciences, Sichuan University)；ZENG Guang-Zhi(Sichuan Huapai Biopharmaceutical company)；WAN Xiao-Ping(Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Key Laboratory for Animal Disease Prevention and Food Safety of Sichuan Province, College of Life Sciences, Sichuan University)；CHEN Yi(Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Key Laboratory for Animal Disease Prevention and Food Safety of Sichuan Province, College of Life Sciences, Sichuan University)；LI Jin-Hai(Sichuan Huapai Biopharmaceutical company)；WANG Ze-Zhou(Center for Animal Disease Control of Sichuan Province)；FANG Pengfei(Sichuan Huapai Biopharmaceutical company)；GAO Rong(Key Laboratory of Bio-resources and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, Key Laboratory for Animal Disease Prevention and Food Safety of Sichuan Province, College of Life Sciences, Sichuan University)
收稿日期：2017-04-19 年卷（期）页码：2017,54(6):1345-1350
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：猪；IL-12基因；免疫；圆环病毒2型疫苗；壳聚糖纳米颗粒
Key words：Pigs; IL-12 gene; immunity; porcine circovirus type 2 vaccine，chitosan nanoparticle
基金项目：国家自然科学基金
中文摘要
为研究白细胞介素-12（IL-12）基因在猪体内表达对圆环病毒2型（PCV2）疫苗免疫应答的调控作用，克隆了藏猪IL-12的P40和P35基因，并将完整的IL-12基因克隆至真核表达载体VR1020，经体外表达验证后，采用离子交联法对藏猪IL-12基因重组质粒进行包裹，制备成粒径小、分散性好且带正电荷的壳聚糖纳米颗粒（VRIL-12-CNP），与PCV2疫苗共同肌肉注射接种21日龄断奶仔猪，在接种后第0天、7天、14天和28天时采集猪前腔静脉血进行血常规分析、流式细胞分析、血清抗体检测和相关免疫基因表达量检测，并记录其生长体重。结果发现：接种了VRIL-12-CNP的实验组猪血液中的CD3+ T细胞、Th细胞、Tc细胞和PCV2抗体在接种后都显著增长（P
英文摘要
To study the regulation effect on vaccination of porcine circovirus-2 (PCV2) by co-delivering interleukin-12 (IL-12) cDNA via chitosan nanoparticles, two subunits of IL-12, P40 and P35 of Tibetan pig, were cloned into mammalian expression plasmid VR1020. VRIL-12 was transfected into HEK293 cells and PCR test was conducted. The results showed that IL-12 gene could express successfully. The recombinant plasmid was packaged into chitosan nanoparticles using ionic crosslinking method. The nanoparticles (VRIL-12-CNP) were confirmed to be nanoparticle size with good dispersion and positive charge. VRIL-12-CNP was then used to inoculate the 21-day-old piglets together with PCV2 vaccine. Blood samples were taken at day 0, 7, 14 and 28 post inoculation for analysis. The results showed that post vaccination the T cells such as CD3+, CD4+ , CD8+, and anti-PCV2 specific antibody, all increased significantly in the VRIL-12-CNP co-delivering pigs (P
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