弓形虫和新孢子虫交叉反应抗原MIC7A对小鼠的免疫保护效力分析

doi:10.11843/j.issn.0366-6964.2022.07.026

畜牧兽医学报 ›› 2022, Vol. 53 ›› Issue (7): 2300-2306.doi: 10.11843/j.issn.0366-6964.2022.07.026

弓形虫和新孢子虫交叉反应抗原MIC7A对小鼠的免疫保护效力分析

陈慧娴¹, 陈雅婕¹, 王先梅¹, 王丽芳¹, 刘群^1,2, 刘晶^1,2*

1. 中国农业大学动物医学院, 国家动物寄生原虫实验室, 北京 100193;
2. 中国农业大学动物医学院, 农业农村部动物流行病学重点实验室, 北京 100193

收稿日期:2021-11-03 出版日期:2022-07-23 发布日期:2022-07-23
通讯作者: 刘晶,主要从事寄生虫病研究,E-mail:liujingvet@cau.edu.cn
作者简介:陈慧娴(1998-),女,浙江台州人,硕士,主要从事寄生虫病诊断研究,E-mail:747843264@qq.com
基金资助:
北京市自然科学基金(6212016)

Identification of the Cross-reacting Antigen MIC17A of Toxoplasma gondii and Neospora caninum and the Study of Its Cross-immune Protective Efficacy in Mice

CHEN Huixian¹, CHEN Yajie¹, WANG Xianmei¹, WANG Lifang¹, LIU Qun^1,2, LIU Jing^1,2*

1. National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China;
2. Key Laboratory of Animal Epidemiology of the Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China

Received:2021-11-03 Online:2022-07-23 Published:2022-07-23

摘要/Abstract

摘要： 弓形虫和新孢子虫是两种亲缘关系接近的顶复亚门原虫，二者之间存在一定程度的交叉免疫保护作用，这种作用可能是基于交叉反应抗原产生的。本研究旨在表达并鉴定弓形虫和新孢子虫的交叉反应抗原TgMIC17A，通过将其应用于小鼠的免疫保护试验，评估该抗原对弓形虫和新孢子虫感染产生的交叉免疫保护作用。对TgMIC17A进行基因克隆和原核表达，通过免疫印迹试验鉴定其反应原性和交叉反应性。重组蛋白免疫小鼠后测定血清特异性IgG抗体水平，评价其免疫原性。二免后，分别用1×10³个弓形虫Pru速殖子、1.5×10⁷个新孢子虫Nc1速殖子攻虫，对小鼠的体重变化、存活率进行监测，并在攻虫30 d后检测各组存活小鼠的脑荷虫量，评价TgMIC17A重组蛋白免疫小鼠后对弓形虫和新孢子虫的交叉免疫保护效果。结果显示，rTgMIC17A可以被弓形虫和新孢子虫的阳性血清识别，相较于未免疫组小鼠，免疫组小鼠体内可产生高水平特异性IgG抗体(P＜0.01)，且感染弓形虫或新孢子虫的脑荷虫量均显著降低(P＜0.01)。本研究克隆并表达了TgMIC17A，鉴定其为新孢子虫和弓形虫的交叉反应抗原。该抗原可以刺激小鼠产生较好的体液免疫反应，并对弓形虫和新孢子虫的感染产生一定的交叉免疫保护作用，可以为弓形虫和新孢子虫共感染的防治，以及筛选具有交叉免疫保护力的重组疫苗提供可借鉴的研究资料。

关键词: MIC17A, 弓形虫, 新孢子虫, 交叉反应抗原, 交叉免疫保护

Abstract: Toxoplasma gondii and Neospora caninum are closely related Apicomplexa parasites. The identified cross-reacting antigens may account for the cross-protection between them. This study aimed to express and identify the cross-reacting antigen TgMIC17A to evaluate the cross immune-protective effect against T. gondii and N. caninum infection in mice. First, TgMIC17A was cloned and expressed in E. coli(DE3), and its cross reactivity and immunogenicity were analyzed by Western blot and ELISA, respectively. Then, mice were immunized with recombinant protein, and challenged with 1×10³ tachyzoites of T. gondii Pru or 1.5×10⁷ N. caninum Nc1 tachyzoites 10 days post the second immunization. The body weight change and survival rate of mice were recorded, and the parasite load in the brain was measured 30 days post-infection to assess the cross-protective efficacies of rTgMIC17A against T. gondii and N. caninum. The results indicated that recombinant TgMIC17A could be recognized by both positive sera of T. gondii and N. caninum. Compared with the unimmunized group, the immunized mice produced high level of specific IgG antibodies (P<0.01), and the amount of T. gondii or N. caninum load in brain significantly decreased as well (P<0.01). In summary, TgMIC17A was identified as a cross-reacting antigen between T. gondii and N. caninum, which can induce robust immune response to protect mice against T. gondii and N. caninum. Taken together, TgMIC17A is a potent cross-protective gene for use as recombinant vaccine against T. gondii or N. caninum infection.

Key words: MIC17A, Toxoplasma gondii, Neospora caninum, cross-reacting antigen, cross-immune protection

中图分类号:

S852.72

陈慧娴, 陈雅婕, 王先梅, 王丽芳, 刘群, 刘晶. 弓形虫和新孢子虫交叉反应抗原MIC7A对小鼠的免疫保护效力分析[J]. 畜牧兽医学报, 2022, 53(7): 2300-2306.

CHEN Huixian, CHEN Yajie, WANG Xianmei, WANG Lifang, LIU Qun, LIU Jing. Identification of the Cross-reacting Antigen MIC17A of Toxoplasma gondii and Neospora caninum and the Study of Its Cross-immune Protective Efficacy in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2300-2306.

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弓形虫和新孢子虫交叉反应抗原MIC7A对小鼠的免疫保护效力分析

Identification of the Cross-reacting Antigen MIC17A of Toxoplasma gondii and Neospora caninum and the Study of Its Cross-immune Protective Efficacy in Mice

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