Interaction between IFITM3 and Foxp3 in mice during Echinococcus multilocularis infections

doi:10.12140/j.issn.1000-7423.2025.04.009

Abstract

Abstract:

Objective To investigate the interaction between interferon-inducible transmembrane protein 3 (IFITM3) and forkhead box P3 (Foxp3) in mice during Echinococcus multilocularis infections. Methods E. multilocularis protoscolices were obtained from gerbil liver tissues, and a C57BL/6J mouse model of E. multilocularis infection was established via intraperitoneal injection with E. multilocularis protoscolices. Mice were euthanized 15, 30, 60 and 90 days post-infection, and mouse liver tissues were collected, with uninfected mice as controls. Pathological changes of mouse liver tissues were observed at different time points using hematoxylin-eosin (HE) staining, Masson staining and immunohistochemistry, and the relative expression of IFITM3 and Foxp3 proteins was determined in liver tissues using Western blotting. HEK-293T cells were transfected with IFITM3-ShRNA interference plasmids (ShIFITM3-1, ShIFITM3-2, ShIFITM3-3), and the optimal knockdown fragments were screened for dose-dependent experiments (transfected at 0, 0.5, 1.0, 1.5, 2.0 μg) to further detect changes in Foxp3 expression. The IFITM3-Foxp3 molecular docking was analyzed using the AutoDock software, and the tag plasmid HA-FITTM3 and Flag-Foxp3a were transfected into HEK-293T cells. Total cellular proteins were extracted for immunoprecipitation to verify the interaction between FITTM3 and Foxp3a. Results HE staining revealed disordered arrangements of liver tissues and gradually aggravated infiltration of inflammatory cells in liver tissues of mice infected with E. multilocularis. Masson staining displayed no significant blue staining areas in the control group, and a gradual increase in the blue staining area in the infection group, with (22.12 ± 3.09)%, (28.57 ± 2.1)%, (41.20 ± 2.01)%, and (58.30 ± 2.21)% proportions 15, 30, 60, and 90 days post-infection, respectively (F = 135.60, P < 0.01). Immunohistochemical staining showed that the positive expression rate of IFITM3 increased from (20.18 ± 7.25)% in the control group to (56.22 ± 4.49)% at 90 days post-infection (F = 25.40, P < 0.05), and the positive expression rate of Foxp3 increased from (2.12 ± 1.49)% in the control group to (59.10 ± 4.45)% at 90 days post-infection (F = 106.30, P < 0.05). Western blotting assay showed that the relative expression level of IFITM3 decreased to 0.27 ± 0.04 at 15 days post-infection, and then increased and reached 1.01 ± 0.05 at 90 days post-infection (F = 52.37, P < 0.05), and the relative expression level of Foxp3 gradually increased over time, from 0.03 ± 0.01 in the control group to 0.97 ± 0.03 at 90 days post-infection (F = 143.20, P < 0.05). The IFITM3 expression positively correlated with Foxp3 expression (R² = 0.687 3, P < 0.05). Transfection with ShIFITM3-1, ShIFITM3-2, and ShIFITM3-3 plasmids all resulted in a decrease in the relative expression level of IFITM3 (0.32 ± 0.02, 0.23 ± 0.05, 0.53 ± 0.09), with ShIFITM3-2 showing the highest knockdown efficiency (F = 37.05, P < 0.01), and the relative expression of Foxp3 gradually decreased with ShIFITM3-2 transfection at 0 (1.10 ± 0.14), 0.5 (0.62 ± 0.05), 1.0 (0.52 ± 0.04), 1.5 (0.49 ± 0.01), and 2.0 μg (0.33 ± 0.05), respectively (F = 42.06, P < 0.01). Molecular docking showed that the docking score between IFITM3 and Foxp3 was -312 kcal/mol, indicating a good affinity, and immunoprecipitation indicated an interaction between IFITM3 and Foxp3. Conclusion There is an interaction between IFITM3 and Foxp3 in mice during E. multilocularis infections, and IFITM3 and Foxp3 expression is positively correlated with the severity of liver damages.

Key words: Echinococcus multilocularis, Interferon-induced transmembrane protein 3, Forkhead box P3, Protein-protein interaction

CLC Number:

R532.32

YUAN Zhen, DIDAER Yeergeming, WANG Fei, JIANG Tao, DUAN Mingjun, AERZIGULI Tuerxun, QI Xinwei, SHAN Jiaoyu. Interaction between IFITM3 and Foxp3 in mice during Echinococcus multilocularis infections[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025, 43(4): 503-510.

Figures/Tables 5

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

References 25

[1]	Wen H, Vuitton L, Tuxun T, et al. Echinococcosis: Advances in the 21st century[J]. Clin Microbiol Rev, 2019, 32(2): e00075.18.
[2]	Woolsey ID, Miller AL. Echinococcus granulosus sensulato and Echinococcus multilocularis: A review[J]. Res Vet Sci, 2021, 135: 517-522.
[3]	韩帅, 李石柱. 我国棘球蚴病流行现状及重点防治工作思考[J]. 中国寄生虫学与寄生虫病杂志, 2025, 43(1): 1-5.
	Han S, Li SZ. Echinococcosis in China: Current status and future disease control priorities[J]. Chin J Parasitol Parasit Dis, 2025, 43(1): 1-5. (in Chinese)
[4]	Gauthiez E, Uldry E, Coste AT, et al. 2023 update on alveolar echinococcosis[J]. Rev Med Suisse, 2023, 19(822): 708-712.
[5]	Vuitton DA, Mantion G, Million L, et al. Alveolar echinococcosis[J]. Rev Prat, 2020, 70(7): 754-764.
[6]	Li XC, Hu XF, You HJ, et al. Regulation of pattern recognition receptor signaling by palmitoylation[J]. iScience, 2025, 28(2): 111667.
[7]	Xie Q, Wang LL, Liao XZ, et al. Research progress into the biological functions of IFITM3[J]. Viruses, 2024, 16(10): 1543.
[8]	Li DY, Wu MH. Pattern recognition receptors in health and diseases[J]. Signal Transduct Target Ther, 2021, 6(1): 291.
[9]	屈晴, 袁振, 地达尔?叶尔革命, 等. 包虫囊液对干扰素信号TRIF/IFITM3通路的影响[J]. 中国病原生物学杂志, 2024, 19(7): 773-778, 783.
	Qu Q, Yuan Z, Didaer Y, et al. The impact of hydatid cyst fluid on the interferon signaling TRIF/IFITM3 pathway[J]. J Pathog Biol, 2024, 19(7): 773-778, 783. (in Chinese)
[10]	Wang J, Gong RN, Zhao CY, et al. Human FOXP3 and tumour microenvironment[J]. Immunology, 2023, 168(2): 248-255.
[11]	Liu SH, Zhang HL, Yan J, et al. FOXP3 and SQSTM1/P62 correlate with prognosis and immune infiltration in hepatocellular carcinoma[J]. Pathol Res Pract, 2023, 242: 154292.
[12]	Dezsényi B, Dubóczki Z, Strausz T, et al. Emerging human alveolar echinococcosis in Hungary (2003-2018): A retrospective case series analysis from a multi-centre study[J]. BMC InfectDis, 2021, 21(1): 168.
[13]	Rodrigues V, Cordeiro-da-Silva A, Laforge M, et al. Impairment of T cell function in parasitic infections[J]. PLoS Negl Trop Dis, 2014, 8(2): e2567.
[14]	Bellanger AP, Mougey V, Pallandre JR, et al. Echinococcus multilocularis vesicular fluid inhibits activation and proliferation of natural killer cells[J]. Folia Parasitol (Praha), 2017, 64: 2017. 029.
[15]	Jiménez-Munguía I, Beaven AH, Blank PS, et al. Interferon-induced transmembrane protein 3 (IFITM3) and its antiviral activity[J]. Current Opinion in Structural Biology, 2022, 77: 102467.
[16]	Lee J, Robinson ME, Ma N, et al. IFITM3 functions as a PIP3 scaffold to amplify PI3K signalling in B cells[J]. Nature, 2020, 588(7838): 491-497.
[17]	Hwang JR, Byeon Y, Kim D, et al. Recent insights of T cell receptor-mediated signaling pathways for T cell activation and development[J]. Exp Mol Med, 2020, 52(5): 750-761.
[18]	Zegeye MM, Lindkvist M, F?lker K, et al. Activation of the JAK/STAT3 and PI3K/AKT pathways are crucial for IL-6 trans-signaling-mediated pro-inflammatory response in human vascular endothelial cells[J]. Cell Commun Signal, 2018, 16(1): 55.
[19]	Xiong ZS, Xu XD, Zhang YX, et al. IFITM3 promotes glioblastoma stem cell-mediated angiogenesis via regulating JAK/STAT3/bFGF signaling pathway[J]. Cell Death Dis, 2024, 15(1): 45.
[20]	Liu XN, Zhang WQ, Han YC, et al. FOXP3⁺ regulatory T cell perturbation mediated by the IFNγ-STAT1-IFITM3 feedback loop is essential for anti-tumor immunity[J]. Nat Commun, 2024, 15(1): 122.
[21]	Liu Z, Lee DS, Liang YQ, et al. Foxp3 orchestrates reorganization of chromatin architecture to establish regulatory T cell identity[J]. Nat Commun, 2023, 14(1): 6943.
[22]	Du GT, Dou CM, Sun P, et al. Regulatory T cells and immune escape in HCC: Un-derstanding the tumor microenvironment and advancing CAR-T cell therapy[J]. Front Immunol, 2024, 15: 1431211.
[23]	Deng G, Song X, Greene MI. FoxP3 in T(reg) cell biology: A molecular and structural perspective[J]. Clin Exp Immunol, 2020, 199(3): 255-262.
[24]	Papadopoulou G, Petroulia S, Karamichali E, et al. The epigenetic controller lysine-specific demethylase 1 (LSD1) regulates the outcome of hepatitis C viral infection[J]. Cells, 2023, 12(21): 2568.
[25]	Liang YM, Li EL, Min JQ, et al. miR-29a suppresses the growth and metastasis of hepatocellular carcinoma through IFITM3[J]. Oncol Rep, 2018, 40(6): 3261-3272.