摘要
为了探究刚地弓形虫组蛋白H2Bb及H2Bv的体外表达及其对小鼠的免疫保护作用,以刚地弓形虫RH株的cDNA作为模板,利用原核表达技术表达弓形虫组蛋白H2Bb及H2Bv。表达后的上述重组蛋白纯化后免疫小鼠,采集免疫后小鼠的血清,测定血清中的总一氧化氮(NO)、抗体及免疫相关细胞因子,分离小鼠脾,测定小鼠脾淋巴细胞的吞噬功能,最后小鼠腹腔接种弓形虫并绘制小鼠的生存曲线。结果显示,成功表达了弓形虫重组蛋白H2Bb(rTgH2Bb)及重组蛋白H2Bv(rTgH2Bv)。Western-blot结果表明重组蛋白具有良好的免疫原性。重组蛋白rTgH2Bb及rTgH2Bv免疫组小鼠血清中Ig G1的水平在一免和二免后均与对照组差异显著(P<0.05),rTgH2Bb及rTgH2Bv免疫组小鼠血清细胞因子γ干扰素的水平在一免及二免后均有显著提高(P<0.05)。rTgH2Bb和rTgH2Bv免疫组小鼠血清中NO含量在2次免疫后均显著高于对照组(P<0.05)。二免后重组蛋白rTgH2Bb免疫组和rTgH2Bv免疫组小鼠脾淋巴细胞的吞噬能力均显著高于对照组(P<0.05)。攻虫感染后,rTgH2Bb和rTgH2Bv免疫组小鼠的生存时间较对照组相比有所延长。结论:重组蛋白rTgH2Bb和rTgH2Bv有一定的抗弓形虫急性感染的作用,具有作为弓形虫疫苗候选抗原的潜在价值。
To explore the expression of Toxoplasma gondii histone H2 Bb and H2 Bv in vitro and their protective efficacy on mice,using the c DNA of T.gondii RH strain as a template,prokaryotic expression technology was used to express T.gondii histone H2 Bb and H2 Bv.The mice were immunized with the purified recombinant proteins.The serum of the immunized mice was collected to detect total nitric oxide(NO),antibody and immune-related cytokines.The spleen of mice was separated and the phagocytosis of spleen lymphocytes was determined.The mice were finally inoculated with T.gondii in the abdominal cavity and the survival curve of the mice was drawn.The recombinant protein H2 Bb(r Tg H2 Bb)and recombinant protein H2 Bv(r Tg H2 Bv)of T.gondii were successfully expressed.Western-blot results showed that the recombinant proteins have good immunogenicity.The levels of Ig G1,cytokines interferonγ(IFN-γ)and NO expression in the serum of mice immunized with recombinant protein r Tg H2 Bb and r Tg H2 Bv were significantly higher than those in the control groups after the first and second immunizations(P<0.05).After the second immunization,the phagocytosis of mice spleen lymphocytes in the r Tg H2 Bb and rTg H2 Bv immunized group was significantly higher than those of the control groups(P<0.05).After challenge infection,the survival time of mice immunized with r Tg H2 Bb and rTg H2 Bv was longer than those of the control groups.In conclusion,recombinant proteins r Tg H2 Bb and r Tg H2 Bv have a certain effect on acute infection of T.gondii,which is conductive to as candidate antigens of toxoplasma vaccine.
作者
周天缘
于正青
李纯静
王茗悦
刘军龙
罗建勋
徐立新
宋小凯
严若峰
李祥瑞
ZHOU Tian-yuan;YU Zheng-qing;LI Chun-jing;WANG Ming-yue;LIU Jun-long;LUO Jian-xun;XU Li-xin;SONG Xiao-kai;YAN Ruo-feng;LI Xiang-rui(MOE Joint International Research Laboratory of Animal Health and Food Safety/College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology of Gansu Province/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2021年第3期296-304,共9页
Chinese Veterinary Science
基金
国家重点基础研究发展计划(973)项目(2015CB150300)
家畜疫病病原生物学国家重点实验室(中国农业科学院兰州兽医研究所)开放课题(SKLVEB2019KFKT009)。
关键词
刚地弓形虫
组蛋白
原核表达
免疫保护
疫苗
Toxoplasma gondii
histone
prokaryotic expression
immune protection
vaccine
