甘露糖化碳纳米管包裹蕨麻多糖作为REV核酸疫苗免疫佐剂的研究  

Study on mannosylated carbon nanotubes encapsulating Potentilla anserine polysaccharide as an immune adjuvant of REV nucleic acid vaccine

作　　者：肖乃依 李杭雨 刘鸿瑞 金圣涵 房笑淞 赵永新 封海波 XIAO Naiyi;LI Hangyu;LIU Hongrui;JIN Shenghan;FANG Xiaosong;ZHAO Yongxin;FENG Haibo(College of Animal Husbandry and Veterinary Medicine,Southwest Minzu University,Chengdu 610041,China)

机构地区：[1]西南民族大学畜牧兽医学院,成都610041

出　　处：《黑龙江畜牧兽医》2024年第16期89-96,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金面上项目(31872511);四川省自然科学基金杰出青年基金项目(2024NSFSC0197);西南民族大学民族兽药研究与开发创新团队项目(2024QGY013);西南民族大学大学生创新创业训练计划项目(S202210656060)。

摘　　要：为了研究甘露糖化碳纳米管包裹蕨麻多糖作为禽网状内皮组织增生症病毒(REV)核酸疫苗免疫佐剂对雏鸡免疫功能的影响,试验利用REV特异性抗原基因gp90与真核表达载体pTT5构建重组质粒pTT5-gp90,提取蕨麻多糖并测定其多糖含量,利用傅里叶红外光谱对甘露糖化碳纳米管进行检测,然后采用CCK-8法测定鸡巨噬细胞(HD11)活性,确定重组质粒pTT5-gp90、蕨麻多糖及甘露糖化碳纳米管使用的质量浓度;将80只未感染REV的1日龄雌性三黄鸡分为对照组、重组质粒pTT5-gp90组、质粒与多糖PAP-gp90组、混合物PAP-gp90@M-M组,分别进行免疫接种、攻毒,免疫后第0,7,14,21天采集血液,检测白细胞介素-4(IL-4)与γ干扰素(IFN-γ)质量浓度、REV抗体效价;攻毒后第7,14,21天饲喂前称量体重并统计累计死亡率,解剖后取脾脏和肝脏,计算脏器指数。结果表明:重组质粒pTT5-gp90、蕨麻多糖、甘露糖化碳纳米管质量浓度分别为100,500,30μg/mL时,HD11细胞活性较高;免疫后第7,14,21,28天,与对照组比较,混合物PAP-gp90@M-M组血清IL-4与IFN-γ质量浓度及REV抗体效价明显上调;攻毒后第3,10天雏鸡体重升高,累计死亡率降低;肝脏指数低于重组质粒pTT5-gp90组,高于对照组,但均差异不显著(P>0.05);脾脏指数显著低于重组质粒pTT5-gp90组(P<0.05),高于对照组(P>0.05)。说明甘露糖化碳纳米管包裹蕨麻多糖可作为REV核酸疫苗的免疫佐剂,该佐剂可通过增强雏鸡免疫力、诱导免疫应答、降低死亡率来提高REV核酸疫苗的免疫效果。In order to study the effects of mannosylated carbon nanotube encapsulating Potentilla anserine polysaccharide as an immune adjuvant for avian Reticuloendotheliosis virus(REV) nucleic acid vaccine on the immune function of chicks, in the experiment, the recombinant plasmid pTT5-gp90 was constructed using the REV-specific antigen gene gp90 and the eukaryotic expression vector pTT5;the Potentilla anserine polysaccharide was extracted and its polysaccharide content was determined;and the mannosylated carbon nanotubes were detected using Fourier infrared spectroscopy. Chicken macrophage(HD11) activity was then determined by CCK-8 assay to determine the used mass concentration of recombinant plasmid pTT5-gp90, Potentilla anserine polysaccharide and mannosylated carbon nanotubes. Eighty 1-day-old female Sanhuang chickens without REV infection were divided into control group, recombinant plasmid pTT5-gp90 group, plasmid and polysaccharide PAP-gp90 group, and mixture PAP-gp90@M-M group. The blood was collected on days 0,7,14 and 21 after immunization, and the mass concentration of interleukin-4(IL-4) and interferon γ(IFN-γ) and the titer of REV antibody were detected. On days 7,14, and 21 after the challenge, the cumulative mortality was measured and weighed before feeding. After dissection, spleen and liver were taken and organ index was calculated. The results showed that HD11 cell activity was higher when the mass concentrations of recombinant plasmid pTT5-gp90, Potentilla anserine polysaccharide, and mannosylated carbon nanotubes were 100, 500, and 30 μg/mL, respectively. On the 7th, 14th, 21st and 28th days after immunization, serum IL-4 and IFN-γ mass concentration and REV antibody titer were significantly up-regulated in the mixture PAP-gp90@M-M group compared with the control group. The chicks' body weight increased and cumulative mortality decreased on the 3rd and 10th days after the challenge;the liver index was lower than that of the recombinant plasmid pTT5-gp90 group and higher than that of the control g

关 键 词：甘露糖化碳纳米管 蕨麻多糖 禽 网状内皮组织增生症病毒 核酸疫苗 免疫佐剂 

分 类 号：S852.4[农业科学—基础兽医学]