BALB/c小鼠感染细粒棘球蚴致敏模型的建立和相关免疫细胞变化的研究

doi:10.12140/j.issn.1000-7423.2020.04.002

摘要/Abstract

摘要：

目的构建BALB/c小鼠感染细粒棘球蚴致敏模型,探讨小鼠相关免疫细胞的变化。方法 18只BALB/c小鼠随机均分为3组,每组6只,分别为致敏组、未致敏组和对照组。将培养的羊源细粒棘球蚴微囊腹腔注射接种感染致敏组和未致敏组小鼠（50个/鼠）,建立细粒棘球蚴感染模型,对照组注射等量生理盐水。感染后6个月,致敏组经腹腔注射细粒棘球蚴粗制囊液致敏,未致敏组和对照组注射等量生理盐水。致敏后,采用症状评分表,每隔5 min对小鼠进行症状评分和肛温测定。致敏后1 h,取各组小鼠内眦静脉血和脾脏,制备脾单细胞悬液。流式细胞术检测小鼠体内树突状细胞（DC）、调节性T细胞（Treg）、辅助性T细胞17（Th17）、白细胞介素-10（IL-10）、转化生长因子-β1（TGF-β1)、IL-17A的变化。采用Graphad Prism 7.0软件进行作图和统计学分析。结果流式细胞术检测结果显示,小鼠DC细胞比例在对照组、未致敏组、致敏组中分别为（3.2 ± 0.5）%、（0.2 ± 0.1）%、（1.5 ± 0.2）%,致敏组DC细胞比例低于对照组（P < 0.01）,高于未致敏组（P < 0.01）。小鼠Treg细胞比例在对照组、未致敏组、致敏组中分别为（5.7 ± 2.1）%、（15.9 ± 3.4）%、（7.4 ± 2.6）%,致敏组低于未致敏组（P < 0.05）。小鼠Th17细胞比例在对照组、未致敏组、致敏组中分别为（4.5 ± 0.6）%、（2.8 ± 0.1）%、（8.6 ± 1.6）%,致敏组高于对照组和未致敏组（P < 0.01）。小鼠IL-10含量在对照组、未致敏组、致敏组中分别为（116.88 ± 15.60）、（1 261.2 ± 208.3）、（386.7 ± 126.5）pg/ml,致敏组高于对照组而低于未致敏组（P < 0.05）。小鼠TGF-β1含量在对照组、未致敏组、致敏组中分别为（36.0 ± 10.2）、（225.9 ± 64.3）、（91.7 ± 15.7）pg/ml,致敏组高于对照组而低于未致敏组（P < 0.05）。小鼠IL-17A含量在对照组、未致敏组、致敏组中分别为（57.3 ± 14.1）、（15.1 ± 1.6）、（168.6 ± 50.6）pg/ml,致敏组高于对照组和未致敏组（P < 0.01）。结论 BALB/c小鼠感染细粒棘球蚴致敏后,Treg细胞数量增多,促进IL-10和TGF-β1的分泌;而DC、Th17细胞数量减少,抑制了IL-17A的产生,从而导致过敏反应。

关键词: 细粒棘球蚴, 过敏反应, 树突状细胞, 调节性T细胞, 辅助性T细胞17

Abstract:

Objective To establish a BALB/c mouse model of Echinococcus granulosus-induced sensitization, and explore the changes of related immune cells.Methods Eighteen BALB/c mice were divided into 3 groups (6 mice each): sensitized group, non-sensitized group and control group. Microcapsules of incubated E. granulosus of sheep source were injected intraperitoneally in mice of the sensitized and non-sensitized groups (50 microcapsules/mouse) to establish the mouse model of E. granulosus infection. The control group received the same volume of normal saline. Six months after infection, mice in the sensitized group was sensitized by intraperitoneal injection of hydatid cyst fluid, while the non-sensitized group and the control group were injected with the same volume of normal saline. After sensitization, the symptoms were scored and anal temperature was measured every 5 min. One hour after sensitization, venous blood of the inner canthus was collected, and the spleen was collected to prepare. Single cell suspension. Flow cytometry was performed to evaluate changes of dendritic cells (DCs), regulatory T cells (Tregs), helper T cells 17 (Th17), interleukin-10 (IL-10), transforming growth factor-β1 (TGF-β1), and IL-17A. Data plotting and analysis were made with the Graphad Prism 7.0 software.Results Results of flow cytometry showed that the percentages of DCs in the control, non-sensitized and sensitized groups were (3.2 ± 0.5)%, (0.2 ± 0.1)% and (1.5 ± 0.2)%, respectively, with that in the sensitized group significantly lower than that in the control group (P < 0.01), but higher than that in the non-sensitized group (P < 0.01). The percentages of Treg cells in the control, non-sensitized and sensitized groups were(5.7 ± 2.1)%, (15.9 ± 3.4)% and (7.4 ± 2.6)%, respectively, with that in the sensitized group significantly lower than that in the non-sensitized group (P < 0.05). The percentages of Th17 cells in the control, non-sensitized and sensitized groups were(4.5 ± 0.6)%, (2.8 ± 0.1)% and(8.6 ± 1.6)%, respectively, with that in the sensitized group significantly higher than that in the control and the non-sensitized groups (P < 0.01). The IL-10 concentration in the control, non-sensitized, and sensitized groups were (116.9 ± 15.6) pg/ml, (1 261.2 ± 208.3) pg/ml, and (386.7 ± 126.5) pg/ml, respectively, with that in the sensitized group significantly higher than that in the control group but lower than that in the non-sensitized group (P < 0.05). The TGF-β1 concentration in the control, non-sensitized, and sensitized groups were (36.0 ± 10.2) pg/ml, (225.9 ± 64.3) pg/ml, and (91.7 ± 15.9) pg/ml, respectively, with that in the sensitized group significantly higher than that in the control group but lower than that in the non-sensitized group (P < 0.05). The IL-17A concentration in the control, non-sensitized, and sensitized groups were (57.3 ± 14.1) pg/ml, (15.1 ± 1.6) pg/ml and (168.6 ± 50.6) pg/ml, respectively, with that in the sensitized group significantly higher than that in the control group and the non-sensitized group (P < 0.01).Conclusion The number of Treg cells was increased after E. granulosus infection in BALB/c mice model, enhancing the secretion of IL-10 and TGF-β1, while the numbers of DC and Th17 cells were decreased, which inhibited the production of IL-17A, leading to the allergic reaction.

Key words: Echinococcus granulosus, Allergic reaction, Dendritic cells, Regulatory T cells, Helper T cells 17

中图分类号:

R532.32

于晓东, 亚力·亚森, 王佳玲, 李孟, 叶建荣. BALB/c小鼠感染细粒棘球蚴致敏模型的建立和相关免疫细胞变化的研究[J]. 中国寄生虫学与寄生虫病杂志, 2020, 38(4): 412-416.

YU Xiao-dong, YALI Ya-sen, WANG Jia-ling, LI Meng, YE Jian-rong. Establishment of BALB/c mouse model of Echinococcus granulosus-induced sensitization and changes of related immune cells[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2020, 38(4): 412-416.

图/表 1

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