Protective effect of IL-4 to Trichinella spiralis infection in the intestinal phase

doi:10.12140/j.issn.1000-7423.2024.03.011

Abstract

Abstract:

Objective To investigate the protective effects of interleukin 4 (IL-4) to Trichinella spiralis infection in the intestinal phase in mice. Methods The muscle samples of T. spiralis reservation mice were digested for collection of muscle larvae, which were sonicated and centrifuged to collect supernatant for preparing Trichinella antigen. Ten wild-type BALB/c mice were randomly divided into the non-infection group and wild-type infection group, with 5 mice each group, while additional 4 IL-4 knockout mice (IL-4KO) were assigned to IL-4KO infection group. The mice of wild-type infection group and IL-4KO infection group were given with 400 muscle larvae each by gavage respectively, while the non-infection group received the same volume of PBS. Eight days after infection, orbital blood samples were obtained to collect sera by centrifugation, and the content of monocyte chemotactic protein-1 (MCP-1) was detected by ELISA. The mice of all groups were dissected to collect the duodenum and proximal jejunum for preparation of paraffin sections. The sections were stained with Hematoxylin and eosin (HE). Measure the length of villi and crypts as well as the number and size of goblet cells useing Aperio ImageScope 12.4.3, and calculate the villus length/crypt length (V/C) and the ratio of the number of goblet cell/villus length (GC/V). Lymphocytes isolated from the mesenteric lymph nodes were cultured with Trichinella antigen (10 μg/ml) for 72 hours, and the supernatant was collected. A Luminex assay was performed to measure the levels of cytokines including IL-1β, IL-12p70, IL-2, IL-5, IL-6, interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α). The data was analyzed using SPSS 26.0 software with independent sample t-tests for pairwise comparisons and one-way ANOVA for multiple sample comparisons. Results ELISA results showed that serum MCP-1 levels were (344.90 ± 21.80), (350.50 ± 38.30) and (467.94 ± 190.01) pg/ml in the non-infection, wild-type infection and IL-4KO infection group, respectively, the IL-4KO infection group was higher than the wild-type infection group (t = 0.681, P < 0.05). HE staining revealed intact mucosa and normal villous architecture with no signs of inflammation in the non-infection group, whereas both the wild-type infection and IL-4KO infection groups showed vacuolated goblet cells, shortened villi and marked inflammation in the duodenum and jejunum, with more severe inflammation in the IL-4KO infection group. The IL-4KO infection group had V/C of 2.62 ± 0.12 and 2.78 ± 0.25 in the duodenum and jejunum, which were both lower than those in the wild-type infection group (3.46 ± 0.05, 3.65 ± 0.12) (F = 24.09, 20.46, P < 0.01, 0.05). The non-infection group had higher V/C of 4.69 ± 0.16 in the jejunum (F = 25.43, P < 0.01). The IL-4KO infection group had GC/V of 9.66 ± 0.88 and 7.33 ± 0.88 in the duodenum and jejunum, which were both higher than those in the wild-type infection group (5.33 ± 1.20 and 4.33 ± 0.33) (F = 17.12, 16.78, both P < 0.05). The goblet cell size in the duodenum and jejunum of the IL-4KO infection group were (12.39 ± 1.17) and (11.05 ± 0.60) μm, both larger than the wild-type infection group, which had (8.33 ± 0.44) and (8.44 ± 0.58) μm (F = 18.47, 16.22, both P < 0.05). The Luminex results showed that the levels of IL-1β, IL-12p70, IL-2, IL-5, IL-6, IFN-γ and TNF-α in the lymphocyte culture supernatants of the IL-4KO infection group were (0.80 ± 0.37), (2.70 ± 0.94), (49.76 ± 16.40), (25.25 ± 3.26), (12.51 ± 4.86), (51.20 ± 8.93), (15.86 ± 2.74) pg/ml, whereas the wild-type infection group had levels of (0.45 ± 0.03), (1.03 ± 0.04), (1.00 ± 0.38), (0.64 ± 0.16), (0.62 ± 0.24), (0.57 ± 0.09), (0.94 ± 0.31) pg/ml, respectively. The IL-4KO infection group were higher than the wild-type infection group (F = 5.52, 24.73, 48.72, 5.00, 123.10, 50.55, P < 0.05 or 0.01) except IL-1β (F = 0.87, P > 0.05). Conclusion IL-4 plays a protective immunological role during the intestinal phase of T. spiralis infection in mice. It could reduce the serum MCP-1, mitigate inflammation response in the duodenum and jejunum and suppress the secretion of inflammatory cytokines.

Key words: Trichinella spiralis, IL-4, Intestinal phase, Infection, Protective effect

CLC Number:

R532.14

LUO Zeni, WU Anqi, WANG Zhikai, PAN Jin, SUN Ximeng. Protective effect of IL-4 to Trichinella spiralis infection in the intestinal phase[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2024, 42(3): 354-359.

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References 27

[1]	Qin YH, Ren YX, Yi C, et al. Effect of wortmannilactone F on Trichinella spiralis enteral in mice[J]. Vector Borne Zoonotic Dis, 2020, 20(3): 205-211.
[2]	Yan JH, Huang SG, Lu FL. Galectin-receptor interactions regulates cardiac pathology caused by Trichinella spiralis infection[J]. Front Immunol, 2021, 12: 639260.
[3]	Bai X, Hu XX, Liu XL, et al. Current research of trichinellosis in China[J]. Front Microbiol, 2017, 8: 1472. doi: 10.3389/fmicb.2017.01472 pmid: 28824597
[4]	Zhang XZ, Yue WW, Bai SJ, et al. Oral immunization with attenuated Salmonella encoding an elastase elicits protective immunity against Trichinella spiralis infection[J]. Acta Trop, 2022, 226: 106263.
[5]	Shimoni Z, Froom P. Uncertainties in diagnosis, treatment and prevention of trichinellosis[J]. Expert Rev Anti Infect Ther, 2015, 13(10): 1279-1288. pmid: 26243167
[6]	Sharma N, Singh V, Shyma KP. Role of parasitic vaccines in integrated control of parasitic diseases in livestock[J]. Vet World, 2015, 8(5): 590-598.
[7]	Zhang X, Sun XM. Research progress on the immune evasion mechanism in Trichinella spiralis infection[J]. Chin J Parasitol Parasit Dis, 2023, 41(4): 492-496, 501. (in Chinese)
	(张旭, 孙希萌. 旋毛虫感染免疫逃逸机制研究进展[J]. 中国寄生虫学与寄生虫病杂志, 2023, 41(4): 492-496, 501.) doi: 10.12140/j.issn.1000-7423.2023.04.016
[8]	Zaini A, Good-Jacobson KL, Zaph C. Context-dependent roles of B cells during intestinal helminth infection[J]. PLoS Negl Trop Dis, 2021, 15(5): e0009340.
[9]	Hou JM, Bo LQ, Wang XY, et al. Study on immune protection of TRE recombinant protein of Trichinella spiralis[J]. Chin Vet Sci, 2023, 53(1): 93-100. (in Chinese)
	(侯嘉茗, 薄禄琪, 王雪莹, 等. 旋毛虫TRE重组蛋白免疫保护作用的研究[J]. 中国兽医科学, 2023, 53(1): 93-100.)
[10]	Hrčková G, Kubašková TM, Reiterová K, et al. Co-administration of silymarin elevates the therapeutic effect of praziquantel through modulation of specific antibody profiles, Th1/Th2/Tregs cytokines and down-regulation of fibrogenesis in mice with Mesocestoides vogae (Cestoda) infection[J]. Exp Parasitol, 2020, 213: 107888.
[11]	Stone KD, Prussin C, Metcalfe DD. IgE, mast cells, basophils, and eosinophils[J]. J Allergy Clin Immunol, 2010, 125(2): S73-S80.
[12]	Gu Y, Wei JF, Yang J, et al. Protective immunity against Trichinella spiralis infection induced by a multi-epitope vaccine in a murine model[J]. PLoS One, 2013, 8(10): e77238.
[13]	Cai J, Huang L, Wang LJ, et al. The role of macrophage polarization in parasitic infections: a review[J]. Chin J Schisto Control, 2020, 32(4): 432-435. (in Chinese)
	(蔡娟, 黄琳, 王灵军, 等. 巨噬细胞极化在寄生虫感染中的作用研究进展[J]. 中国血吸虫病防治杂志, 2020, 32(4): 432-435.)
[14]	Chen YF, Zheng JJ, Qu C, et al. Inonotus obliquus polysaccharide ameliorates dextran sulphate sodium induced colitis involving modulation of Th1/Th2 and Th17/Treg balance[J]. Artif Cells Nanomed Biotechnol, 2019, 47(1): 757-766.
[15]	van Dyken SJ, Locksley RM. Interleukin-4- and interleukin-13-mediated alternatively activated macrophages: roles in homeostasis and disease[J]. Annu Rev Immunol, 2013, 31: 317-343. doi: 10.1146/annurev-immunol-032712-095906 pmid: 23298208
[16]	Liang HE, Reinhardt RL, Bando JK, et al. Divergent expression patterns of IL-4 and IL-13 define unique functions in allergic immunity[J]. Nat Immunol, 2011, 13(1): 58-66.
[17]	Chen F, Liu ZG, Wu WH, et al. An essential role for Th2-type responses in limiting acute tissue damage during experimental helminth infection[J]. Nat Med, 2012, 18(2): 260-266. doi: 10.1038/nm.2628 pmid: 22245779
[18]	Herbert DR, Hölscher C, Mohrs M, et al. Alternative macrophage activation is essential for survival during schistosomiasis and downmodulates T helper 1 responses and immunopathology[J]. Immunity, 2004, 20(5): 623-635. doi: 10.1016/s1074-7613(04)00107-4 pmid: 15142530
[19]	Cai QW, Zhao JJ, Li HL, et al. Detection of serum IL-4 and IL-10 in patients with Taeniasis asiatica[J]. Henan J Prev Med, 2022, 33(10): 804-806. (in Chinese)
	(蔡倩文, 赵俊杰, 李海龙, 等. 亚洲带绦虫病患者血清IL-4和IL-10含量检测[J]. 河南预防医学杂志, 2022, 33(10): 804-806.)
[20]	Singh S, Anshita D, Ravichandiran V. MCP-1: function, regulation, and involvement in disease[J]. Int Immunopharmacol, 2021, 101(Pt B): 107598.
[21]	Yoshimura T. The chemokine MCP-1 (CCL2) in the host interaction with cancer: a foe or ally?[J]. Cell Mol Immunol, 2018, 15(4): 335-345. doi: 10.1038/cmi.2017.135 pmid: 29375123
[22]	Liu Y, Zhang S, Luo Z, et al. Supplemental Bacillus subtilis PB6 improves growth performance and gut health in broilers challenged with Clostridium perfringens[J]. J Immunol Res, 2021, 2021: 2549541.
[23]	Fariña FA, Pasqualetti MI, Bessi C, et al. Reprint of: comparison between Trichinella patagoniensis and Trichinella spiralis infection in BALB/c mice[J]. Vet Parasitol, 2021, 297: 109542.
[24]	Guo K, Sun XM, Gu Y, et al. Trichinella spiralis paramyosin activates mouse bone marrow-derived dendritic cells and induces regulatory T cells[J]. Parasit Vectors, 2016, 9(1): 569.
[25]	Knight PA, Wright SH, Lawrence CE, et al. Delayed expulsion of the nematode Trichinella spiralis in mice lacking the mucosal mast cell-specific granule chymase, mouse mast cell protease-1[J]. J Exp Med, 2000, 192(12): 1849-1856. doi: 10.1084/jem.192.12.1849 pmid: 11120781
[26]	Urban JF Jr, Schopf L, Morris SC, et al. Stat6 signaling promotes protective immunity against Trichinella spiralis through a mast cell- and T cell-dependent mechanism[J]. J Immunol, 2000, 164(4): 2046-2052. doi: 10.4049/jimmunol.164.4.2046 pmid: 10657657
[27]	Finkelman FD, Shea-Donohue T, Morris SC, et al. Interleukin-4- and interleukin-13-mediated host protection against intestinal nematode parasites[J]. Immunol Rev, 2004, 201: 139-155. pmid: 15361238