Establishment of BALB/c mouse model of Echinococcus granulosus-induced sensitization and changes of related immune cells

doi:10.12140/j.issn.1000-7423.2020.04.002

Abstract

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

CLC Number:

R532.32

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.

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References 21

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