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壳聚糖纳米颗粒包裹猪白细胞介素2和融合白细胞介素4/6基因对猪圆环病毒2型疫苗免疫的强化
Enhancement of Immune Responses to PCV-2 Vaccine by Porcine IL-2 and Fusion IL-4/6 Gene Entrapped in Chitosan Nanoparticles
陈祎1#, 宋婷玉1#, 李金海2#, 肖永乐1, 曾光志2, 万小平1, 杨璐一1, 方鹏飞2*, 王泽洲3*, 高荣1*
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作者单位:1. 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, 成都 610065;
2. 四川省华派生物制药有限公司, 成都 610026;
3. 四川省动物疫病预防控制中心, 成都 610035
中文关键字:仔猪;基因共表达;白细胞介素2;融合白细胞介素4/6;免疫;PCV-2疫苗
英文关键字:piglet; gene co-expression; IL-2; fusion IL-4/6; immunity; PCV-2 vaccine
中文摘要:目的 探讨猪Sus scrofa domesticus白细胞介素2(IL-2)和融合白细胞介素4/6(IL-4/6)基因的共表达对仔猪免疫应答的影响,为进一步研制新型免疫调节剂来加强仔猪抵御断奶后多系统衰竭综合征奠定基础。方法 将猪IL-2和IL-4/6融合基因的共表达重组质粒,用壳聚糖材料包裹制成纳米颗粒,记作VRIL-4/6-2-CS。将仔猪分为2组,分别肌肉注射VRIL-4/6-2-CS(实验组)和生理盐水(对照组),2组均同时接种猪圆环病毒2型(PCV-2)疫苗,在接种后的第0、7、14、28天采集血样并分析免疫变化。结果 实验组仔猪血清中的IgG2a抗体数量和血液中CD4+、CD8+ T淋巴细胞数量均显著高于对照组(P<0.05);同时,实验组仔猪的IL-2、IL-4、IL-6、TNF-α、TLRs(TLR-2,7)、STATs(STAT-1,2,3)基因表达水平也显著高于对照组(P<0.05)。尽管2组之间PCV-2特异性抗体的含量差异无统计学意义,但是实验组仔猪的生长速率显著高于对照组(P<0.05)。结论 VRIL-4/6-2-CS能促进仔猪对PCV-2疫苗的免疫应答,是一种安全有效的免疫佐剂。
英文摘要:Objective In order to develop a novel effective immunomodulator to enhance pig resistance against post-weaning multisystemic wasting syndrome, we used the recombinant plasmid co-expressing the pig interleukin-2 (IL-2) and fusion interleukin-4/6 (IL-4/6) proteins which we constructed before, and studied its effect on immune response of piglets. Methods The recombinant plasmid was first encapsulated in chitosan nanoparticles by the ionotropic gelation method, and generated VRIL-4/6-2-CS. Then piglets were divided into 2 groups and intramuscularly injected with VRIL-4/6-2-CS or saline followed by the injection of pig circovirus-2 (PCV-2) vaccine, respectively. The blood was collected from each piglet on days 0, 7, 14 and 28 to assay the immunological changes. Result The number of IgG2a, CD4+, CD8+ T cells increased significantly in the sera or blood of piglets that treated with VRIL-4/6-2-CS (P<0.05). Furthermore, the expression levels of IL-2, IL-4, IL-6, TNF-α, TLRs (TLR-2, 7) and STATs (STAT-1, 2, 3) genes elevated significantly in the piglets that treated with VRIL-4/6-2-CS (P<0.05). Although no significant differences were observed in the levels of PCV-2-specific antibody, the growth performance of VRIL-4/6-2-CS treated piglets was remarkably improved in comparison with that of control (P<0.05). Conclusion VRIL-4/6-2 entrapped in chitosan is a promising effective adjuvant to promote the immune responses of pig vaccinated with PCV-2.
2018,37(2): 156-163 收稿日期:2017-03-30
DOI:10.11984/j.issn.1000-7083.20170102
分类号:Q78
基金项目:国家自然科学基金项目(30871855);科技部国际合作项目(2011DFA10101103)
作者简介:陈祎(1993-),女,硕士研究生,研究方向:分子遗传与基因工程,E-mail:chenyizoe@163.com;宋婷玉(1992-),女,硕士研究生,研究方向:分子遗传与基因工程,E-mail:stytiki@163.com;李金海(1976-),男,博士,研究方向:动物生物技术,E-mail:ljh76429@sina.com
*通讯作者:高荣,E-mail:gaorong96@163.com
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