Changes of splenic type 1 innate lymphoid cells subsets and their related factors expression in mice infected with Echinococcus multilocularis

doi:10.12140/j.issn.1000-7423.2026.02.003

Abstract

Abstract:

Objective To investigate the effects of Echinococcus multilocularis infection on the proportion of type 1 innate lymphoid cells (ILC1) and the expression levels of related factors in the spleen of mice. Methods C57BL/6J mice were randomly divided into the control group and the infection group. The infection group mice were injected with 3 000 E. multilocularis protoscoleces via the portal vein, while the control group mice were injected with the same volume of normal saline. After 24 weeks, spleen tissue sections were prepared from the infection group and the control group mice, and hematoxylin-eosin (HE) staining was used to observe the pathological changes in the spleen. Splenic lymphocytes were isolated from the mice, and flow cytometry was used to detect the proportion of ILC1 cell subsets, the expression of immune checkpoint molecules, and the levels of secreted cytokines in the spleen. Results HE staining showed that the boundary between the red pulp and the marginal zone of the white pulp was blurred in the spleen of mice infected with E. multilocularis, and hemosiderin deposition occurred in the white pulp area. Flow cytometry results indicated that the percentage of splenic natural killer (NK) cells among total splenic lymphocytes in the control group was (2.52 ± 0.44)%, which was higher than that in the infection group (1.25 ± 0.28)% (t = 6.42，P < 0.01). The percentage of splenic ILC1 among splenic NK cells in the control group was (20.07 ± 2.70)%, which was lower than that in the infection group (31.49 ± 3.28)% (t = 7.10, P < 0.01). The percentage of splenic conventional NK (cNK) cells among splenic NK cells in the infection group was (68.89 ± 3.28)%, which was lower than that in the control group (80.14 ± 3.49)% (t = 6.22, P < 0.01). The percentage of CD27^-CD11b⁺ cells among splenic ILC1 in the infection group was (21.37 ± 7.48)%, which was lower than that in the control group (43.49 ± 8.05)% (t = 5.33, P < 0.01); the percentages of CD27⁺CD11b^- and CD27⁺CD11b⁺ cells were (39.64 ± 8.80)% and (6.77 ± 1.51)%, respectively, which were higher than those in the control group (24.51 ± 8.35)% and (4.12 ± 1.71)% (t = 3.30, 3.06; both P < 0.01). The percentages of splenic ILC1 secreting programmed death receptor 1 (PD-1) and glucocorticoid-induced tumor necrosis factor receptor (GITR) in the infection group were (5.04 ± 1.54)% and (16.14 ± 4.53)%, respectively, which were higher than those in the control group (2.57 ± 1.52)% and (10.81 ± 4.03)% (t = 3.02, 2.33; both P < 0.05). The percentages of splenic ILC1 secreting granzyme B (Gr-B) and interferon-γ (IFN-γ) in the infection group were (6.16 ± 1.23)% and (16.13 ± 6.64)%, respectively, which were lower than those in the control group (9.42 ± 2.16)% and (30.68 ± 6.81)% (t = 2.94, 3.42; P < 0.05, 0.01). The percentage of splenic ILC1 secreting tumor necrosis factor-α (TNF-α) in the infection group was (22.26 ± 6.69)%, which was higher than that in the control group (10.80 ± 3.92)% (t = 3.31, P < 0.05). Conclusion In the late stage of E. multilocularis infection, the proportion of ILC1 cells and immature ILC1 cells were increased in mouse spleen. The co-inhibitory immune checkpoint molecule PD-1 was highly expressed, and the ability of these cells to secrete cytokines Gr-B and IFN-γ decreased. This suggested that ILC1 cell function suppressed might promote the chronic parasitism of E. multilocularis.

Key words: Echinococcus multilocularis, Alveolar echinococcosis, Spleen, NK cells, ILC1, Immune checkpoint molecules

CLC Number:

R532.32

XIAO Wenying, ABIDAN Ainiwaer, GE Conghui, TANG Na, SUN Sheng, WANG Mengying, GAO Yi, AYINAER Jiensi, HU Qiu, LI Jing, WANG Hui, ZHANG Chuanshan. Changes of splenic type 1 innate lymphoid cells subsets and their related factors expression in mice infected with Echinococcus multilocularis[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2026, 44(2): 166-173.

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References 30

[1]	Carmena D, Benito A, Eraso E. The immunodiagnosis of Echinococcus multilocularis infection[J]. Clin Microbiol Infect, 2007, 13(5): 460-475.
[2]	Davidson RK, Romig T, Jenkins E, et al. The impact of globalisation on the distribution of Echinococcus multilocularis[J]. Trends Parasitol, 2012, 28(6): 239-247.
[3]	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.
[4]	施阳, 李德伟, 阿比旦?艾尼瓦尔, 等. 多房棘球蚴感染对小鼠脾脏自然杀伤细胞Tim3表达的影响[J]. 中国血吸虫病防治杂志, 2023, 35(4): 366-373.
	Shi Y, Abidan ANWE, Li DW, et al. Effect of Echinococcus multilocularis infection on Tim3 expression in spleen natural killer cells of mice[J]. Chin J Schisto Control, 2023, 35(4): 366-373. (in Chinese)
[5]	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)
[6]	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)
[7]	王宝辉. 脾脏CD49b^-NK细胞的发育分化研究[D]. 合肥: 中国科学技术大学, 2019: 46-51.
	Wang BH. The development and differentiation of spleen CD49b^-NK cells[D]. Hefei: University of Science and Technology of China, 2019: 46-51. (in Chinese)
[8]	Ding Y, Lavaert M, Grassmann S, et al. Distinct developmental pathways generate functionally distinct populations of natural killer cells[J]. Nat Immunol, 2024, 25(7): 1183-1192.
[9]	Shannon MJ, Mace EM. Natural killer cell integrins and their functions in tissue residency[J]. Front Immunol, 2021, 12: 647358.
[10]	Chen SM, Zhu HT, Jounaidi Y. Comprehensive snapshots of natural killer cells functions, signaling, molecular mechanisms and clinical utilization[J]. Signal Transduct Target Ther, 2024, 9: 302.
[11]	Gasteiger G, Fan XY, Dikiy S, et al. Tissue residency of innate lymphoid cells in lymphoid and nonlymphoid organs[J]. Science, 2015, 350(6263): 981-985.
[12]	Sojka DK, Plougastel-Douglas B, Yang LP, et al. Tissue-resident natural killer (NK) cells are cell lineages distinct from thymic and conventional splenic NK cells[J]. eLife, 2014, 3: e01659.
[13]	Sivori S, Pende D, Quatrini L, et al. NK cells and ILCs in tumor immunotherapy[J]. Mol Aspects Med, 2021, 80: 100870.
[14]	Michel T, Poli A, Domingues O, et al. Mouse lung and spleen natural killer cells have phenotypic and functional differences, in part influenced by macrophages[J]. PLoS One, 2012, 7(12): e51230.
[15]	Taggenbrock RLRE, ILC1: development, maturation, and transcriptional regulation[J]. Eur J Immunol, 2023, 53(2): 2149435.
[16]	Zhang CS, Shao YM, Yang ST, et al. T-cell tolerance and exhaustion in the clearance of Echinococcus multilocularis: role of inoculum size in a quantitative hepatic experimental model[J]. Sci Rep, 2017, 7: 11153.
[17]	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.
[18]	Crane GM, LiuYC, Chadburn A. Spleen: development, anatomy and reactive lymphoid proliferations[J]. Semin Diagn Pathol, 2021, 38(2): 112-124.
[19]	Cenariu D, Iluta S, Zimta AA, et al. Extramedullary hematopoiesis of the liver and spleen[J]. J Clin Med, 2021, 10(24): 5831.
[20]	Gutiérrez-Hoya A, Soto-Cruz I. NK cell regulation in cervical cancer and strategies for immunotherapy[J]. Cells, 2021, 10(11): 3104.
[21]	Stojanovic A, Cerwenka A. ILC1-like NK cells as matchmakers for DC-T cell interactions[J]. Immunity, 2021, 54(10): 2185-2187.
[22]	Van Acker N, Frenois FX, Gravelle P, et al. Spatial mapping of innate lymphoid cells in human lymphoid tissues and lymphoma at single-cell resolution[J]. Nat Commun, 2025, 16: 4545.
[23]	He JM, Chen DL, Xiong W, et al. Eomesodermin spatiotemporally orchestrates the early and late stages of NK cell development by targeting KLF2 and T-bet, respectively[J]. Cell Mol Immunol, 2024, 21(7): 662-673.
[24]	Samper N, Hardardottir L, Depierreux DM, et al. Kir6.1, a component of an ATP-sensitive potassium channel, regulates natural killer cell development[J]. Front Immunol, 2024, 15: 1490250.
[25]	Flommersfeld S, B?ttcher JP, Ersching J, et al. Fate mapping of single NK cells identifies a type 1 innate lymphoid-like lineage that bridges innate and adaptive recognition of viral infection[J]. Immunity, 2021, 54(10): 2288-2304. e7.
[26]	Lu JS, Hu ZQ, Jiang HJ, et al. Dual nature of type I interferon responses and feedback regulations by SOCS1 dictate malaria mortality[J]. J Adv Res, 2025, 73: 295-310.
[27]	Montes de Oca M, Kumar R, de Labastida Rivera F, et al. Type I interferons regulate immune responses in humans with blood-stage Plasmodium falciparum infection[J]. Cell Rep, 2016, 17(2): 399-412.
[28]	Hu Y, Wang XL, Wei YH, et al. Functional inhibition of natural killer cells in a BALB/c mouse model of liver fibrosis induced by Schistosoma japonicum infection[J]. Front Cell Infect Microbiol, 2020, 10: 598987.
[29]	Zhou J, Peng H, Li K, et al. Liver-resident NK cells control antiviral activity of hepatic T cells via the PD-1-PD-L1 axis[J]. Immunity, 2019, 50(2): 403-417. e4.
[30]	Abulizi A, Shao YM, Aji T, et al. Echinococcus multilocularis inoculation induces NK cell functional decrease through high expression of NKG2A in C57BL/6 mice[J]. BMC Infect Dis, 2019, 19(1): 792.