一种山羊支原体山羊肺炎亚种多重抗原肽的小鼠免疫试验

doi:10.11843/j.issn.0366-6964.2016.12.018

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (12): 2476-2482.doi: 10.11843/j.issn.0366-6964.2016.12.018

一种山羊支原体山羊肺炎亚种多重抗原肽的小鼠免疫试验

赵萍，陈胜利，郝华芳，李学瑞，贺英，储岳峰^*，刘永生^*

（中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部草食动物疫病重点开放实验室，兰州 730046）

收稿日期:2016-06-15 出版日期:2016-12-23 发布日期:2016-12-23
通讯作者: 储岳峰， E-mail:chuyuefeng@caas.cn；刘永生，E-mail:liuyongsheng@caas.cn
作者简介:赵萍(1973-)，女，甘肃会宁人，副研究员，硕士，主要从事细菌分子生物学， E-mail:zhaoping73@126.com，Tel:0931-8342676
基金资助:
国家科技支撑计划（2015BAD12B02）；十三五项目“牛羊重要病原体分子检测新技术研究”（2016YFD0500907）

Immune Responses in Mice Immunized with a Recombinant Multiple Antigenic Peptide from Mycoplasma capricolum Subsp. capripneumoniae

ZHAO Ping, CHEN Sheng-li，HAO Hua-fang, LI Xue-rui, HE Ying, CHU Yue-feng^*, LIU Yong-sheng^*

（State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Epizootic Diseases of Grazing Animals of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China）

Received:2016-06-15 Online:2016-12-23 Published:2016-12-23

摘要/Abstract

摘要：

旨在构建一种由山羊支原体山羊肺炎亚种(Mycoplasma capricolum Subsp. capripneumoniae，Mccp)六个B细胞表位和三个T细胞表位组成的多重抗原肽(multiple antigenic peptide，MAP)，将其在大肠杆菌中表达后免疫小鼠，评估其体液免疫和细胞免疫。分别用MAP、单个B细胞表位及全菌超破抗原作为抗原，检测免疫小鼠抗体水平，结果显示三种抗原均能和小鼠免疫血清发生很好的反应(P<0.01)。体外代谢抑制试验显示，1∶64倍稀释的免疫小鼠血清可以有效抑制Mccp的生长。淋巴细胞增殖试验显示，当用单个T细胞表位及Mccp超破抗原分别刺激免疫小鼠的淋巴细胞时均产生明显的增殖现象(P<0.01)。细胞因子检测结果显示，免疫小鼠的IFN-γ、TNF-α、IL-1β和 IL-10产量明显升高(P<0.001)，而IL-12产量明显下降(P<0.001)。数据显示，构建的MAP能够引起小鼠的免疫反应，这一结果为由Mccp引起的山羊传染性胸膜肺炎亚单位疫苗的研制奠定了一定的基础。

关键词: 免疫反应, 小鼠, 多重抗原肽, 山羊支原体山羊肺炎亚种

Abstract:

A multiple antigenic peptide (MAP), composed of six B-cell epitopes and three T-cell epitopes derived from antigenic proteins of Mycoplasma capricolum subsp. capripneumoniae (Mccp), was designed and expressed in Escherichia coli cells, which was then injected into mice to evaluate humoral and cellular immune responses induced by the recombinant MAP. Antibody levels against MAP, each single B-cell epitope, and the ultrasonic-treated crude Mccp antigen were all significantly increased (P<0.01). Results of in vitro metabolic inhibition test indicated that diluted sera (1:64) from immunized mice inhibited Mccp growth. Splenic lymphocytes from the immunized mice exhibited remarkable proliferation when stimulated by each single T-cell epitope and crude Mccp antigen (P<0.01). The production of IFN-γ, TNF-α, IL-1β, and IL-10 were increased (P<0.001), while that of IL-12 was decreased (P<0.001). These data showed that the constructed MAP stimulated immune responses in mice. The results laid a certain foundation to develop subunit vaccine against contagious caprine pleuropneumonia caused by Mccp.

Key words: immune response, mice, multiple antigenic peptides, Mycoplasma capricolum subsp. capripneumoniae

中图分类号:

S852.62

赵萍，陈胜利，郝华芳，李学瑞，贺英，储岳峰，刘永生. 一种山羊支原体山羊肺炎亚种多重抗原肽的小鼠免疫试验[J]. 畜牧兽医学报, 2016, 47(12): 2476-2482.

ZHAO Ping, CHEN Sheng-li，HAO Hua-fang, LI Xue-rui, HE Ying, CHU Yue-feng, LIU Yong-sheng. Immune Responses in Mice Immunized with a Recombinant Multiple Antigenic Peptide from Mycoplasma capricolum Subsp. capripneumoniae[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(12): 2476-2482.

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一种山羊支原体山羊肺炎亚种多重抗原肽的小鼠免疫试验

Immune Responses in Mice Immunized with a Recombinant Multiple Antigenic Peptide from Mycoplasma capricolum Subsp. capripneumoniae

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