CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >

2025 , Vol. 43 >Issue 1: 76 - 83

DOI: https://doi.org/10.12140/j.issn.1000-7423.2025.01.012

ORIGINAL ARTICLES

Immune cell migration and its functional changes in the peritoneal cavity of mice infected with Echinococcus multilocular

  • DENG Bingqing ,
  • ADILAI Duolikun ,
  • LI Yinshi ,
  • ABIDAN Ainiwaer ,
  • SUN Sheng ,
  • XIAO Wenying ,
  • GE Conghui ,
  • TANG Na ,
  • JIANG Tao ,
  • WANG Hui ,
  • ZHANG Chuanshan ,
  • LI Jing
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  • 1 College of Basic Medical, Xinjiang Medical University, Urumqi 830054, Xinjiang, China
    2 Clinical Medicine Research Institute, the First Afilicated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang, China
    3 Xinjiang Medical University Laboratory Animal Center, Urumqi 830011, Xinjiang, China
    4 Key Laboratory of High Incidence Disease Research in Xingjiang (Xinjiang Medical University), Ministry of Education, Urumqi 830054, Xinjiang, China
    5 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention (Chinese Center for Tropical Diseases Research); NHC Key Laboratory of Parasite and Vector Biology (National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention), Shanghai 200025, China

Received date: 2024-10-21

  Revised date: 2025-01-24

  Online published: 2025-03-26

Supported by

Natural Science Foundation of Xinjiang Uyghur Autonomous Region(2022D01E51);Natural Science Foundation of Xinjiang Uyghur Autonomous Region(2022D01D60);National Natural Science Foundation of China(82302561);National Natural Science Foundation of China(82372279);National Natural Science Foundation of China(82160396);Funding of Open Subjects in the Key Laboratory of the Ministry of Education for the Study of Highly Prevalent Diseases in the Xinjiang Region(2023B03);Provincial-Ministerial Joint Establishment of State Key Laboratory of Causes and Prevention of High Morbidity in Central Asia(SKL-HIDCA-2024-25);Open Topic Funding Program of the Key Laboratory of Parasitic Pathogen and Vector Biology of the National Health Commission of China(NHCKFKT2022-05)

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Abstract

Objective To understand the changes in the migration and differentiation of immune cells in the peritoneal cavity of mice infected with Echinococcus multilocularis, and to explore the mechanism of immune function. Methods Spleen tissues of enhanced green fluorescent protein (EGFP) mice were lysed to isolate EGFP-labeled immune cells. C57BL/6J mice were intraperitoneally injected with 2.5 × 10⁷ traced immune cells. Five days later, 3 000 protoscoleces of E. multilocularis were inoculated via the hepatic portal vein. After 19 weeks of infection, the liver and spleen tissues of the infected mice were collected. After frozen sectioning, the sections were stained with 4,6-diamidino-2-phenylindole (DAPI), and immunofluorescence was used to observe the distribution of the traced immune cells. Immune cells from peripheral blood, liver, spleen and peritoneal cavity of infected mice were isolated, and the migration distribution and cell typing of EGFP-labeled immune cells in different tissues were detected by flow cytometry, as well as the secretion of cytokines interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) and interleukin-4 (IL-4). Results The immunofluorescence results showed that the EGFP-traced cells mainly migrated to the spleen tissue of the mice. In the liver tissue, the proportions of various types of traced lymphocytes from high to low were as follows: CD8+ T cells (79.63 ± 5.00)%, CD4+ T cells (7.08 ± 4.00)%, and B cells (2.01 ± 1.00)% (F = 396.20, P < 0.01). In the spleen tissue, the proportions of various types of lymphocytes from high to low were as follows: CD8+ T cells (73.97 ± 3.56)%, CD4+ T cells (11.92 ± 5.02)% and B cells (3.86 ± 0.32)% (F = 349.00, P < 0.01). In the peritoneal cavity, the proportions of various types of lymphocytes from high to low were as follows: CD8+ T cells (45.27 ± 4.53)%, CD4+ T cells (34.60 ± 8.00)% and B cells (15.03 ± 6.38)% (F = 16.90, P < 0.05). The numbers of traced CD4+ T cells in the liver, spleen, and peritoneal cavity were 2 700 ± 3 120, 14 407 ± 11 958 and 3 376 ± 1 481 respectively. The numbers of traced CD8+ T cells were 26 407 ± 22 190, 109 179 ± 92 692 and 4 245 ± 798 respectively. The numbers of traced B cells were 698 ± 719, 5 794 ± 4 971 and 1 476 ± 840 respectively. All three types of traced lymphocytes were mainly distributed in the spleen tissue (F = 2.51, 3.03, 2.62, all P > 0.05). In the liver tissue, the proportions of various types of myeloid cells from high to low were as follows: dendritic cells (3.94 ± 1.33)%, monocytes (3.79 ± 1.80)%, macrophages (3.13 ± 1.85)%, eosinophils (2.40 ± 1.81)%, myeloid-derived suppressor cells (0.76 ± 0.17)% and neutrophils (0.67 ± 0.04)% (F = 3.21, P < 0.05). In the spleen tissue, the proportions of various types of myeloid cells from high to low were as follows: dendritic cells (1.86 ± 0.89)%, monocytes (0.41 ± 0.09)%, macrophages (0.23 ± 0.03)%, neutrophils and myeloid-derived suppressor cells [both (0.13 ± 0.14)%], and eosinophils (0.04 ± 0.04)% (F = 42.70, P < 0.01). In the peripheral blood, the proportions of various types of myeloid cells from high to low were as follows: dendritic cells (0.37 ± 0.28)%, monocytes (0.22 ± 0.27)%, macrophages (0.22 ± 0.21)%, neutrophils and myeloid-derived suppressor cells [both (0.17 ± 0.08)%], and eosinophils (0.05 ± 0.08)% (F = 0.92, P > 0.05). Among the traced CD4+ T cells in the liver tissue, the cells secreting IFN-γ, TNF-α, and IL-4 accounted for (41.17 ± 19.92)%, (30.70 ± 3.35)%, and (2.78 ± 4.81)% respectively (F = 8.22, P < 0.05). Among the traced CD8+ T cells, the cells secreting IFN-γ, TNF-α, and IL-4 accounted for (64.17 ± 3.17)%, (24.85 ± 15.66)%, and (5.67 ± 6.35)% respectively (F = 27.08, P < 0.01). Among the traced CD4+ T cells in the spleen tissue, the cells secreting IFN-γ, TNF-α, and IL-4 accounted for (52.43 ± 19.43)%, (27.67 ± 18.99)%, and (4.77 ± 2.23)% respectively (F = 6.88, P < 0.05). Among the traced CD8+ T cells, the cells secreting TNF-α, IFN-γ, and IL-4 accounted for (40.53 ± 17.79)%, (38.97 ± 18.04)%, and (3.23 ± 3.09)% respectively (F = 6.15, P < 0.05). Among the traced CD4+ T cells in the peritoneal cavity, the cells secreting IFN-γ, TNF-α, and IL-4 accounted for (64.33 ± 6.82)%, (46.43 ± 13.44)%, and (4.03 ± 3.51)% respectively (F = 36.04, P < 0.01). Among the traced CD8+ T cells, the cells secreting IFN-γ, TNF-α, and IL-4 accounted for (53.57 ± 19.17)%, (21.47 ± 8.54)%, and (6.13 ± 8.63)% respectively (F = 10.24, P < 0.05). Conclusion Immune cells in the peritoneal cavity of E. multilocularis infected mice mainly migrate to the spleen tissue, and the cell types are mainly CD8+ T cells and dendritic cells. T cells mainly secreted IFN-γ, which showed Th1 immune response.

Key words： Alveolar echinococcoisis; Peritoneal cavity; Immume cell; Migration; CD8+ T cell

Cite this article

DENG Bingqing , ADILAI Duolikun , LI Yinshi , ABIDAN Ainiwaer , SUN Sheng , XIAO Wenying , GE Conghui , TANG Na , JIANG Tao , WANG Hui , ZHANG Chuanshan , LI Jing . Immune cell migration and its functional changes in the peritoneal cavity of mice infected with Echinococcus multilocular[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025 , 43(1) : 76 -83 . DOI: 10.12140/j.issn.1000-7423.2025.01.012

References

[1] Alvi MA, Alsayeqh AF. Food-borne zoonotic echinococcosis: A review with special focus on epidemiology[J]. Front Vet Sci, 2022, 9: 1072730.
[2] Wen H, Vuitton L, Tuxun T, et al. Echinococcosis: Advances in the 21st century[J]. Clin Microbiol Rev, 2019, 32(2): e00075-18.
[3] Peters L, Jiang WJ, Eberhardt N, et al. 18FDG-PET/CT-scans and biomarker levels predicting clinical outcome in patients with alveolar echinococcosis: A single-center cohort study with 179 patients[J]. Pathogens, 2023, 12(8): 1041.
[4] Torgerson PR, Schweiger A, Deplazes P, et al. Alveolar echinococcosis: From a deadly disease to a well-controlled infection. Relative survival and economic analysis in Switzerland over the last 35 years[J]. J Hepatol, 2008, 49(1): 72-77.
[5] Zhang CS, Wang H, Li J, et al. Involvement of TIGIT in natural killer cell exhaustion and immune escape in patients and mouse model with liver Echinococcus multilocularis infection[J]. Hepatology, 2021, 74(6): 3376-3393.
[6] Gaudino SJ, Kumar P. Cross-talk between antigen presenting cells and T cells impacts intestinal homeostasis, bacterial infections, and tumorigenesis[J]. Front Immunol, 2019, 10: 360.
[7] 刘志刚, 张万华, 刘霆. 腹膜腔细胞的免疫特性及功能研究进展[J]. 重庆医学, 2019, 48(14): 2467-2470.
  Liu ZG, Zhang WH, Liu T. Advances in research on immune characteristics and function of peritoneal cells[J]. Chongqing Med, 2019, 48(14): 2467-2470. (in Chinese)
[8] Huntington ND, Gray DH. Immune homeostasis in health and disease[J]. Immunol Cell Biol, 2018, 96(5): 451-452.
[9] Wagner PL, Knotts CM, Donneberg VS, et al. Characterizing the immune environment in peritoneal carcinomatosis: Insights for novel immunotherapy strategies[J]. Ann Surg Oncol, 2024, 31(3): 2069-2077.
[10] 吕群燕, 田志刚, 董尔丹, 等. 组织器官的区域免疫特性与疾病机理研究[J]. 科学通报, 2012, 57(36): 3450-3458.
  Lü QY, Tian ZG, Dong ED, et al. Tissue/organ-specialized immunologic features and disease mechanisms[J]. Chin Sci Bull, 2012, 57(36): 3450-3458. (in Chinese)
[11] Lodygin D, Flügel A. Intravital real-time analysis of T-cell activation in health and disease[J]. Cell Calcium, 2017, 64: 118-129.
[12] Mejri N, Gottstein B. Intraperitoneal Echinococcus multilocularis infection in C57BL/6 mice affects CD40 and B7 costimulator expression on peritoneal macrophages and impairs peritoneal T cell activation[J]. Parasite Immunol, 2006, 28(8): 373-385.
[13] 李建辉, 彭心宇, 周宗瑶, 等. 骨桥蛋白在肝细粒棘球蚴外囊壁中的表达[J]. 中国寄生虫学与寄生虫病杂志, 2006, 24(3): 171-174.
  Li JH, Peng XY, Zhou ZY, et al. Expression of osteopontin in the pericystic layer of hepatic hydatid cyst[J]. Chin J Parasitol Parasit Dis, 2006, 24(3): 171-174. (in Chinese)
[14] Wu FR, Pan CX, Rong C, et al. Inhibition of acid-sensing ion channel 1a in hepatic stellate cells attenuates PDGF-induced activation of HSCs through MAPK pathway[J]. Mol Cell Biochem, 2014, 395(1/2): 199-209.
[15] Kilwinski J, Jenne L, Jellen-Ritter A, et al. T lymphocyte cytokine profile at a single cell level in alveolar echinococcosis[J]. Cytokine, 1999, 11(5): 373-381.
[16] Manfras BJ, Reuter S, Wendland T, et al. Increased activation and oligoclonality of peripheral CD8+ T cells in the chronic human helminth infection alveolar echinococcosis[J]. Infect Immun, 2002, 70(3): 1168-1174.
[17] Mejri N, Müller N, Hemphill A, et al. Intraperitoneal Echinococcus multilocularis infection in mice modulates peritoneal CD4+ and CD8+ regulatory T cell development[J]. Parasitol Int, 2011, 60(1): 45-53.
[18] 侯娇, 温浩, 王明坤, 等. 多房棘球蚴感染小鼠脾脏巨噬细胞亚群及其极化表型的变化[J]. 中国寄生虫学与寄生虫病杂志, 2021, 39(6): 771-778.
  Hou J, Wen H, Wang MK, et al. Changes of macrophage subsets and polarization in spleen of mice infected with Echinococcus multilocularis[J]. Chin J Parasitol Parasit Dis, 2021, 39(6): 771-778. (in Chinese)
[19] 侯昕伶, 李亮, 李玲慧, 等. 泡球蚴感染对小鼠脾脏CD8+ T细胞免疫功能耗竭的影响[J]. 中国血吸虫病防治杂志, 2020, 32(6): 591-597, 604.
  Hou XL, Li L, Li LH, et al. Exhaustion of CD8+ T cell immune functions in spleen of mice with different doses of Echinococcus multilocularis infections[J]. Chin J Schisto Control, 2020, 32(6): 591-597, 604. (in Chinese)
[20] 张伶慧, 陈根, 种世桂, 等. 多房棘球蚴病中免疫细胞调控机制的研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2022, 40(1): 109-113, 120.
  Zhang LH, Chen G, Chong SG, et al. Research progress on the immune regulation mechanism in alveolar echinococcosis[J]. Chin J Parasitol Parasit Dis, 2022, 40(1): 109-113, 120. (in Chinese)
[21] 徐凯, 王海久, 张丽, 等. 多房棘球蚴对宿主肝细胞损害机制的研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2021, 39(2): 256-260.
  Xu K, Wang HJ, Zhang L, et al. Research progress on the mechanisms underlying the impairment of host hepatocytes by Echinococcus multilocularis[J]. Chin J Parasitol Parasit Dis, 2021, 39(2): 256-260. (in Chinese)
[22] Grubor NM, Jovanova-Nesic KD, Shoenfeld Y. Liver cystic echinococcosis and human host immune and autoimmune follow-up: A review[J]. World J Hepatol, 2017, 9(30): 1176-1189.
[23] Zhang CS, Lin RY, Li ZD, et al. Immune exhaustion of T cells in alveolar echinococcosis patients and its reversal by blocking checkpoint receptor TIGIT in a murine model[J]. Hepatology, 2020, 71(4): 1297-1315.
[24] Schmid M, Samonigg H, St?ger H, et al. Use of interferon gamma and mebendazole to stop the progression of alveolar hydatid disease: Case report[J]. Clin Infect Dis, 1995, 20(6): 1543-1546.
[25] Liance M, Ricard-Blum S, Emery I, et al. Echinococcus multilocularis infection in mice: In vivo treatment with a low dose of IFN-gamma decreases metacestode growth and liver fibrogenesis[J]. Parasite, 1998, 5(3): 231-237.
[26] 章宁. γ-干扰素在泡球蚴感染小鼠中的作用研究[D]. 乌鲁木齐: 新疆医科大学, 2020: 1-8.
  Zhang N. Study on the effect of interferon-γ in mice with Echinococcus multilocularis infection[D]. Urumqi: Xinjiang Medical University, 2020: 1-8. (in Chinese)
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