多房棘球蚴感染小鼠外周血中髓源抑制性细胞比例动态变化及细胞因子表达研究

doi:10.12140/j.issn.1000-7423.2024.02.012

中国寄生虫学与寄生虫病杂志 ›› 2024, Vol. 42 ›› Issue (2): 211-216.doi: 10.12140/j.issn.1000-7423.2024.02.012

多房棘球蚴感染小鼠外周血中髓源抑制性细胞比例动态变化及细胞因子表达研究

苏雅馨¹(), 江楠¹, 章孝成¹, 王莹¹, 蒋小凤¹, 霍乐乐¹, 王雅雪¹, 曹建平¹^,², 沈玉娟¹^,²^,^*()

1 中国疾病预防控制中心寄生虫病预防控制所（国家热带病研究中心），传染病溯源预警与智能决策全国重点实验室，国家卫生健康委员会寄生虫病原与媒介生物学重点实验室，世界卫生组织热带病合作中心，科技部国家级热带病国际研究中心，上海 200025
2 上海交通大学医学院-国家热带病研究中心全球健康学院，上海 200025

收稿日期:2024-02-28 修回日期:2024-03-26 出版日期:2024-04-30 发布日期:2024-04-28
通讯作者: * 沈玉娟（1969—），女，硕士，研究员，从事棘球蚴感染与免疫、药物靶点及作用机制，以及肠道原虫分子流行病学、溯源和检测技术等研究。E-mail：shenyj@nipd.chinacdc.cn
作者简介:苏雅馨（1995—），女，硕士研究生，从事棘球蚴感染与免疫研究。E-mail：syx01251228@163.com
基金资助:
国家自然科学基金(82372283);国家自然科学基金(82072307)

Study on the dynamic change of myeloid-derived suppressor cells proportion and cytokine expression in peripheral blood of mice infected with Echinococcus multilocularis

SU Yaxin¹(), JIANG Nan¹, ZHANG Xiaocheng¹, WANG Ying¹, JIANG Xiaofeng¹, HUO Lele¹, WANG Yaxue¹, CAO Jianping¹^,², SHEN Yujuan¹^,²^,^*()

1 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; Chinese Center for Tropical Diseases Research; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases; Key Laboratory on Parasite and Vector Biology, Ministry of Health; WHO Collaborating Centre for Tropical Diseases; National Center for International Research on Tropical Diseases, Ministry of Science and Technology, Shanghai 200025, China
2 School of Global Health, Chinese Center for Tropical Diseases Research, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

Received:2024-02-28 Revised:2024-03-26 Online:2024-04-30 Published:2024-04-28
Contact: * E-mail: shenyj@nipd.chinacdc.cn
Supported by:
National Natural Science Foundation of China(82372283);National Natural Science Foundation of China(82072307)

摘要/Abstract

摘要：

目的分析多房棘球蚴原头节感染小鼠外周血白细胞中髓源抑制性细胞（MDSC）及其亚型的比例动态变化，以及感染晚期小鼠血清中与MDSC增殖相关细胞因子的表达情况。方法将24只BALB/c小鼠随机分为感染组和对照组，每组12只。感染组小鼠经腹腔注射1 200个原头节，对照组小鼠注射等量生理盐水。每组小鼠分别于感染后30、90和180 d随机各取3只，观察肝、脾组织形态学变化，并采集眼眶静脉丛外周血制备白细胞。外周血白细胞用外标抗体CD11b、Gr-1、Ly6G和Ly6C孵育后，流式细胞仪检测MDSC及其亚型多核MDSC（PMN-MDSC）和单核MDSC（M-MDSC）的占比。感染后180 d，采集感染组和对照组小鼠血清，ELISA检测白细胞介素6（IL-6）、IL-13、肿瘤坏死因子α（TNF-α）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）的浓度。使用GraphPad Prism 9.0软件进行制图与统计学分析，两组间比较采用独立样本t检验。结果感染多房棘球蚴原头节后30 d，感染组小鼠肝组织出现囊泡，囊泡的体积随感染时间延长而增加；脾组织随感染时间延长逐渐肿大。流式细胞分析结果显示，感染后30、90和180 d，感染组小鼠外周血白细胞中MDSC占比分别为（13.2 ± 2.4）%、（15.7 ± 2.3）%和（41.3 ± 4.0）%，对照组小鼠的占比分别为（12.4 ± 3.2）%、（6.0 ± 0.9）%和（22.3 ± 1.1）%，感染后90和180 d感染组均高于对照组（t = 3.949、4.682，P < 0.05、0.01）；感染后30、90和180 d，感染组小鼠外周血白细胞中PMN-MDSC占比分别为（10.9 ± 2.1）%、（12.5 ± 2.4）%和（35.8 ± 3.6）%，对照组小鼠的占比分别为（9.6 ± 3.1）%、（4.5 ± 0.6）%和（18.5 ± 0.6）%，感染后90和180 d感染组均高于对照组（t = 3.237、4.788，P < 0.05、0.01）；感染后30、90和180 d，感染组小鼠外周血白细胞中M-MDSC占比分别为（1.8 ± 0.3）%、（1.1 ± 0.1）%和（4.6 ± 1.1）%，对照组小鼠的占比分别为（2.3 ± 0.2）%、（0.5 ± 0.1）%和（3.4 ± 0.9）%，感染后90 d感染组高于对照组（t = 3.246，P < 0.05）。感染后180 d，感染组小鼠血清中IL-6、IL-13、TNF-α和GM-CSF的浓度分别为（315.39 ± 13.58）、（339.41 ± 13.35）、（223.53 ± 27.49）和（262.31 ± 2.36） pg/ml，均高于对照组的（14.93 ± 0.55）、（50.74 ± 0.88）、（50.64 ± 1.64）和（115.28 ± 0.58） pg/ml（t = 22.100、21.580、6.277、60.460，P < 0.01或0.05）。结论感染多房棘球蚴原头节后，小鼠外周血白细胞中MDSC比例随时间延长而增加，以PMN-MDSC增加为主。感染晚期小鼠血清中IL-6、IL-13、TNF-α和GM-CSF浓度升高，可能促进MDSC的增殖与活化。

关键词: 多房棘球蚴, 原头节, 髓源抑制性细胞, 细胞因子

Abstract:

Objective To analyze the dynamic changes in the proportion of myeloid-derived suppressor cells (MDSCs) and their subtypes in peripheral blood leukocytes of mice infected with Echinococcus multilocularis protoscoleces and detect the expression of cytokines related to MDSCs proliferation in the late-infected mice serum. Methods Twenty-four BALB/c mice were randomly divided into the infected group and the control group, with 12 mice in each group. Mice in the infected group were intraperitoneally injected with 1 200 E. multilocularis protoscoleces, while mice in the control group injected with the same volume of normal saline. Three mice from each group were randomly selected at 30, 90 and 180 days after infection to observe histomorphological changes in liver and spleen. Peripheral blood was collected from orbital venous plexus to prepare peripheral blood leukocytes. After incubation with externally labeled antibodies to CD11b, Gr-1, Ly6G and Ly6C, flow cytometry was used to detect the proportions of MDSCs, polymorphonuclear MDSCs (PMN-MDSCs) and monocytic MDSCs (M-MDSCs). Serum samples from both groups were collected 180 days after infection for determine the concentrations of interleukin-6 (IL-6), IL-13, tumor necrosis factor α (TNF-α) and granulocyte-macrophage colony-stimulating factor (GM-CSF) by ELISA assay. GraphPad Prism 9.0 software was utilized for graphical and statistical analysis; independent sample t-test was used for inter-group comparison. Results After E. multilocularis protoscoleces infection, cysts appeared in the liver of infected group mice 30 days after infection, and volume of cysts was increased over time. Spleen of infected group mice were enlaged over time 30, 90, and 180 days after infection. The results of flow cytometry showed that 30, 90 and 180 days after infection, the proportion of MDSCs in peripheral blood leukocytes of infected group were (13.2 ± 2.4)%, (15.7 ± 2.3)% and (41.3 ± 4.0)%, respectively. And the proportion of MDSCs in the control group were (12.4 ± 3.2)%, (6.0 ± 0.9)% and (22.3 ± 1.1)%, respectively. The infected group was significantly higher than the control group 90 and 180 days after infection (t = 3.949, 4.682; P < 0.05, 0.01). 30, 90 and 180 days after infection, the proportion of PMN-MDSCs in peripheral blood leukocytes of infected mice were (10.9 ± 2.1)%, (12.5 ± 2.4)% and (35.8 ± 3.6)%, respectively. And the proportion of PMN-MDSCs in the control group were (9.6 ± 3.1)%, (4.5 ± 0.6)% and (18.5 ± 0.6)%, respectively. The infected group was significantly higher than the control group 90 and 180 days after infection (t = 3.237, 4.788, P < 0.05, 0.01). 30, 90 and 180 days after infection, the proportion of M-MDSCs in peripheral blood leukocytes of infected mice were (1.8 ± 0.3)%, (1.1 ± 0.1)% and (4.6 ± 1.1)%, respectively. And the proportion of M-MDSCs in the control group were (2.3 ± 0.2)%, (0.5 ± 0.1)% and (3.4 ± 0.9)%, respectively. The infected group was significantly higher than the control group 90 days after infection (t = 3.246, P < 0.05). 180 days after infection, the concentrations of IL-6, IL-13, TNF-α and GM-CSF in the infected mice serum were (315.39 ± 13.58), (339.41 ± 13.35), (223.53 ± 27.49) and (262.31 ± 2.36) pg/ml, respectively, which were significantly higher than that of control group [(14.93 ± 0.55), (50.74 ± 0.88), (50.64 ± 1.64) and (115.28 ± 0.58) pg/ml] (t = 22.100, 21.580, 6.277, 60.460; P < 0.01, 0.01, 0.05, 0.01). Conclusion The proportion of MDSCs in peripheral blood leukocytes increased with time after E. multilocularis protoscoleces infection, mainly of PMN-MDSCs. The concentrations of serum IL-6, IL-13, TNF-α and GM-CSF increased of mice in the late stage of E. multilocularis protoscoleces infection, which may promote the proliferation and activation of MDSCs.

Key words: Echinococcus multilocularis, Protoscoleces, Myeloid-derived suppressor cell, Cytokine

中图分类号:

R532.32

苏雅馨, 江楠, 章孝成, 王莹, 蒋小凤, 霍乐乐, 王雅雪, 曹建平, 沈玉娟. 多房棘球蚴感染小鼠外周血中髓源抑制性细胞比例动态变化及细胞因子表达研究[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(2): 211-216.

SU Yaxin, JIANG Nan, ZHANG Xiaocheng, WANG Ying, JIANG Xiaofeng, HUO Lele, WANG Yaxue, CAO Jianping, SHEN Yujuan. Study on the dynamic change of myeloid-derived suppressor cells proportion and cytokine expression in peripheral blood of mice infected with Echinococcus multilocularis[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2024, 42(2): 211-216.

图/表 2

参考文献 23

[1]	Wen H, Vuitton L, Tuxun T, et al. Echinococcosis: advances in the 21st century[J]. Clin Microbiol Rev, 2019, 32(2): e00075.
[2]	World Health Organization. World health statistics 2018: monitoring health for the SDGs, sustainable development goals[M]. Geneva: WHO, 2018: 6-8.
[3]	Jenkins EJ, Schurer JM, Gesy KM. Old problems on a new playing field: helminth zoonoses transmitted among dogs, wildlife, and people in a changing northern climate[J]. Vet Parasitol, 2011, 182(1): 54-69.
[4]	Wang JH, Müller S, Lin RY, et al. Depletion of FoxP3⁺ Tregs improves control of larval Echinococcus multilocularis infection by promoting co-stimulation and Th1/17 immunity[J]. Immun Inflamm Dis, 2017, 5(4): 435-447.
[5]	Marvel D, Gabrilovich DI. Myeloid-derived suppressor cells in the tumor microenvironment: expect the unexpected[J]. J Clin Invest, 2015, 125(9): 3356-3364.
[6]	Condamine T, Mastio J, Gabrilovich DI. Transcriptional regulation of myeloid-derived suppressor cells[J]. J Leukoc Biol, 2015, 98(6): 913-922.
[7]	Gabrilovich DI. Myeloid-derived suppressor cells[J]. Cancer Immunol Res, 2017, 5(1): 3-8.
[8]	Pan W, Zhou HJ, Shen YJ, et al. Surveillance on the status of immune cells after Echinnococcus granulosus protoscoleces infection in BALB/c mice[J]. PLoS One, 2013, 8(3): e59746.
[9]	Yu AP, Yin JH, Gong WC, et al. Changes of myeloid-derived suppressor cells and Th17 cells in mice infected with Echinococcus granulosus protoscoleces[J]. Chin J Parasitol Parasit Dis, 2018, 36(3): 224-230. (in Chinese)
	(于爱萍, 尹建海, 巩文词, 等. 细粒棘球蚴感染小鼠髓源抑制性细胞与Th17细胞比例的变化[J]. 中国寄生虫学与寄生虫病杂志, 2018, 36(3): 224-230.)
[10]	Veglia F, Sanseviero E, Gabrilovich DI. Myeloid-derived suppressor cells in the era of increasing myeloid cell diversity[J]. Nat Rev Immunol, 2021, 21(8): 485-498.
[11]	Jou E, Rodriguez-Rodriguez N, Ferreira AF, et al. An innate IL-25-ILC2-MDSC axis creates a cancer-permissive microenvironment for Apc mutation-driven intestinal tumorigenesis[J]. Sci Immunol, 2022, 7(72): eabn0175.
[12]	Groth C, Hu XY, Weber R, et al. Immunosuppression mediated by myeloid-derived suppressor cells (MDSCs) during tumour progression[J]. Br J Cancer, 2019, 120(1): 16-25.
[13]	?zkan B, Lim H, Park SG. Immunomodulatory function of myeloid-derived suppressor cells during B cell-mediated immune responses[J]. Int J Mol Sci, 2018, 19(5): 1468.
[14]	Ma TM, Renz BW, Ilmer M, et al. Myeloid-derived suppressor cells in solid tumors[J]. Cells, 2022, 11(2): 310.
[15]	Condamine T, Ramachandran I, Youn JI, et al. Regulation of tumor metastasis by myeloid-derived suppressor cells[J]. Annu Rev Med, 2015, 66: 97-110.
[16]	Safarzadeh E, Orangi M, Mohammadi H, et al. Myeloid-derived suppressor cells: important contributors to tumor progression and metastasis[J]. J Cell Physiol, 2018, 233(4): 3024-3036.
[17]	Cao SK, Pan W, Liu H, et al. Expression and activity of arginase from monocytic-type myeloid-derived suppressor cells in rats infected with Echinococcus granulosus[J]. Chin J Parasitol Parasit Dis, 2016, 34(1): 27-31. (in Chinese)
	(曹胜魁, 潘伟, 刘华, 等. 细粒棘球蚴感染小鼠单核髓源抑制性细胞精氨酸酶的表达和活性研究[J]. 中国寄生虫学与寄生虫病杂志, 2016, 34(1): 27-31.)
[18]	Zhang N, Zhang CS, Li ZD, et al. The effects of Echinococcus multilocularis protoscoleces infection on lymphocytes and their subpopulations in mouse liver and spleen[J]. Chin J Parasitol Parasit Dis, 2020, 38(1): 30-35. (in Chinese)
	(章宁, 张传山, 李智德, 等. 多房棘球蚴感染对小鼠肝脾淋巴细胞及其亚群的影响[J]. 中国寄生虫学与寄生虫病杂志, 2020, 38(1): 30-35.)
[19]	Zhang XF, Gong WC, Cao SK, et al. Dynamic changes of myeloid-derived suppressor cells and regulatory T cells in livers of mice infected with Echinococcus granulosus[J]. Chin J Schisto Control, 2019, 31(6): 622-627. (in Chinese)
	(张小凡, 巩文词, 曹胜魁, 等. 细粒棘球绦虫感染小鼠肝脏髓源抑制性细胞与调节性T细胞比例动态变化[J]. 中国血吸虫病防治杂志, 2019, 31(6): 622-627.)
[20]	Weber R, Riester Z, Hüser L, et al. IL-6 regulates CCR5 expression and immunosuppressive capacity of MDSC in murine melanoma[J]. J Immunother Cancer, 2020, 8(2): e000949.
[21]	Weber R, Groth C, Lasser S, et al. IL-6 as a major regulator of MDSC activity and possible target for cancer immunotherapy[J]. Cell Immunol, 2021, 359: 104254.
[22]	Grover A, Sanseviero E, Timosenko E, et al. Myeloid-derived suppressor cells: a propitious road to clinic[J]. Cancer Discov, 2021, 11(11): 2693-2706.
[23]	Petrova V, Groth C, Bitsch R, et al. Immunosuppressive capacity of circulating MDSC predicts response to immune checkpoint inhibitors in melanoma patients[J]. Front Immunol, 2023, 14: 1065767.

多房棘球蚴感染小鼠外周血中髓源抑制性细胞比例动态变化及细胞因子表达研究

Study on the dynamic change of myeloid-derived suppressor cells proportion and cytokine expression in peripheral blood of mice infected with Echinococcus multilocularis

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 2

参考文献 23

相关文章 15

编辑推荐

Metrics

本文评价

[1]	邓冰清, 阿迪莱·多力坤, 李寅时, 阿比旦·艾尼瓦尔, 孙胜, 肖雯颖, 葛聪蕙, 唐娜, 姜涛, 王慧, 张传山, 李静. 多房棘球蚴感染小鼠腹膜腔免疫细胞迁移及其功能变化[J]. 中国寄生虫学与寄生虫病杂志, 2025, 43(1): 76-83.
[2]	魏盼龙, 张耀刚, 张涛, 杨紫晗, 侯静, 田美媛, 黄登亮, 马艳艳. 多房棘球蚴感染巨噬细胞中CD47和SIRPα表达变化与抑制作用的研究[J]. 中国寄生虫学与寄生虫病杂志, 2025, 43(1): 84-90.
[3]	李振伟, 王成, 王志鑫, 刘津铭, 赵乾, 王海久, 谢智. 基于三维可视化技术指导肝多房棘球蚴病手术方案的应用研究[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(6): 737-743.
[4]	祖力皮喀尔·图孙尼亚孜, 蒋铁民, 温浩. 肝多房棘球蚴病的手术治疗进展[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(6): 783-789.
[5]	尹先敏, 种世桂, 陈根, 秦俊梅, 赵玉敏. 巨噬细胞调控多房棘球蚴病所致肝纤维化研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(6): 790-795.
[6]	李嘉静, 黄文君, 曹得萍. 壮药鸡骨草制剂治疗多房棘球蚴感染诱导的肝纤维化效果[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(5): 573-581.
[7]	赵涵玥, 李锦田, 薛俊隆, 卡力比夏提·艾木拉江, 林仁勇, 吐尔干艾力·阿吉. 多房棘球蚴感染晚期小鼠肝脏中NK1.1的表达对T细胞功能的影响[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(5): 594-600.
[8]	徐刚, 毛艺, 李江, 张宏伟, 张永国, 吴向未, 彭心宇, 孙红, 杨婧, 陈骞, 张示杰. 多房棘球蚴通过肝脏p38MAPK通路调控棘球蚴自身生长[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(4): 447-453.
[9]	葛宇飞, 徐刚, 张宏伟, 李江, 张永国, 孙红, 杨婧, 张示杰. 全反式维甲酸对多房棘球蚴原头节体外活性及生长的影响[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(3): 303-308.
[10]	蒉嫣, 薛垂召, 王旭, 刘白雪, 王莹, 王立英, 杨诗杰, 韩帅, 许学年. 2022年全国棘球蚴病防治工作进展[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(1): 8-16.
[11]	曹得萍, 李嘉静, 宋梦微, 莫刚. 多房棘球蚴组织蛋白体外刺激肝星状细胞变化的实验观察[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(4): 440-445.
[12]	朱爱娅, 王旭, 王江友, 王颖, 李杨, 宋珊, 耿燕, 兰子尧, 戴佳芮. 贵州省儿童多房棘球蚴病1例[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(4): 520-523.
[13]	娆琬·托勒洪, 阿不都撒拉木·阿不力克木, 杨凌菲, 陈璐, 李钊, 贾芳, 宋涛. 超声表现诊断肝多房棘球蚴病的效果评价及因素分析[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(3): 312-318.
[14]	蒉嫣, 薛垂召, 王旭, 刘白雪, 王莹, 王立英, 杨诗杰, 韩帅, 伍卫平, 肖宁. 2021年全国棘球蚴病防治进展[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(2): 142-148.
[15]	郭刚, 任远, 焦红杰, 武娟, 郭宝平, 齐文静, 李军, 张文宝. 腹腔感染棘球蚴微囊对小鼠肝脏多房棘球蚴感染及致病的影响[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(2): 156-162.