产肠毒素大肠杆菌感染IPEC-J2细胞后IL-17细胞因子表达变化及其功能初探

doi:10.11843/j.issn.0366-6964.2019.04.016

Abstract

Abstract:

Enterotoxigenic E. coli (ETEC) is one of the most important cause of neonatal and post-weaning diarrhea (PWD) in piglets. The interaction between ETEC and intestinal epithelial cells is the prerequisite of bacterial pathogenesis, as well as the first step of triggering host immune response. The aim of this study was to determine the expression profile and host defense mechanisms of IL-17 cytokine in porcine intestinal epithelial cells against ETEC infection. Firstly, we determined the expression of IL-17 cytokines and several mucosal defense genes by the application of RT-PCR, ELISA, immunohistochemistry (IHC) assays and Western Blotting in the porcine intestinal epithelial cell line IPEC-J2 infected with a F4⁺ETEC reference strain C83901. Later on, the regulating effect of IL-17 cytokine on the expression of above mucosal defense genes in IPEC-J2 cells was explored. The results showed that C83901 induced the mRNA expression of all IL-17 cytokines (IL-17D was not detected), particularly upregulated the expression of IL-17A and IL-17C both at the transcriptional and protein level. The upregulation of IL-17A and IL-17C in the protein level were confirmed by IHC and ELISA. The existence of IL-17A and/or IL-17C either induced by ETEC infection or supplemented artificially in IPEC-J2 cells can elicit the expression of host defense gene expression of mucin-2, claudin-1, claudin-2 and beta defensins-2. These results indicated that IL-17 induction in intestinal epithelial cells upon ETEC infection are beneficial for the mucosal host defense by promoting the intestinal barrier integrity.

CLC Number:

S852.612

LUO Yu, XU Jia, ZHANG Chaoying, JIANG Chunyan, HE Haijian, YU Jianguo, ZHANG Hongbing. The Expression and Functional Analysis of IL-17 Cytokine Family in Enterotoxigenic E.coli Infected IPEC-J2 Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(4): 830-839.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2019.04.016

https://www.xmsyxb.com/EN/Y2019/V50/I4/830

References

[1] 张秀媛,袁万哲,孙继国. 产肠毒素性大肠杆菌主要毒力因子及其疫苗的研究进展[J]. 畜牧与兽医, 2014, 46(5):112-116.
ZHANG X Y, YUAN W Z, SUN J G. Research progress on main virulence factors and vaccines of enterotoxigenic Escherichia coli[J]. Animal Husbandry & Veterinary Medicine, 2014, 46(5):112-116. (in Chinese)
[2] GYLES C L, PRESCOTT J F, SONGER G, et al. Pathogenesis of bacterial infections in animals[M]. 4th ed. Ames, Iowa:Wiley-Blackwell, 2010.
[3] PETERSON L W, ARTIS D. Review. Intestinal epithelial cells:regulators of barrier function and immune homeostasis[J]. Nat Rev Immunol, 2014, 14(3):141-153.
[4] BROSNAHAN A J, BROWN D R. Porcine IPEC-J2 intestinal epithelial cells in microbiological investigations[J]. Vet Microbiol, 2012, 156(3-4):229-237.
[5] ZHOU C L, LIU Z Z, JIANG J C, et al. Differential gene expression profiling of porcine epithelial cells infected with three enterotoxigenic Escherichia coli strains[J]. BMC Genomics, 2012, 13:330
[6] GAO Y, HAN F, HUANG X, et al. Changes in gut microbial populations, intestinal morphology, expression of tight junction proteins, and cytokine production between two pig breeds after challenge with Escherichia coli K88:a comparative study[J]. J Anim Sci, 2013, 91(12):5614-5625.
[7] NGENDAHAYO MUKIZA C, DUBREUIL J D.Escherichia coli heat-stable toxin b impairs intestinal epithelial barrier function by altering tight junction proteins[J]. Infect Immun, 2013, 81(8):2819-2827.
[8] NASSOUR H, DUBREUIL J D.Escherichia coli STb enterotoxin dislodges claudin-1 from epithelial tight junctions[J]. PLoS One, 2014, 9(11):e113273.
[9] KUMAR P, LUO Q W, VICKERS T J, et al.EatA, an immunogenic protective antigen of enterotoxigenic Escherichia coli, degrades intestinal mucin[J]. Infect Immun, 2014, 82(2):500-508.
[10] LUO Q W, KUMAR P, VICKERS T J, et al. Enterotoxigenic Escherichia coli secretes a highly conserved mucin-degrading metalloprotease to effectively engage intestinal epithelial cells[J]. Infect Immun, 2014, 82(2):509-521.
[11] ZHOU M, ZHU J, YU H, et al. Investigation into in vitro and in vivo models using intestinal epithelial IPEC-J2 cells and Caenorhabditis elegans for selecting probiotic candidates to control porcine enterotoxigenic Escherichia coli[J]. J Appl Microbiol, 2014, 117(1):217-226.
[12] SPITZER F, SPEISER S, VAHJEN W, et al. Effect of different feed ingredients and additives on IPEC-J2 cells challenged with an enterotoxigenic Escherichia coli strain[J]. Cytotechnology, 2016, 68(4):1463-1471.
[13] KARIMI S, JONSSON H, LUNDH T, et al.Lactobacillus reuteri strains protect epithelial barrier integrity of IPEC-J2 monolayers from the detrimental effect of enterotoxigenic Escherichia coli[J]. Physiol Rep, 2018, 6(2):e13514.
[14] KERN M, GÜNZEL D, ASCHENBACH J R, et al. Altered cytokine expression and barrier properties after in vitro infection of porcine epithelial cells with enterotoxigenic Escherichia coli and probiotic Enterococcus faecium[J]. Mediators Inflamm, 2017, 2017:2748192.
[15] SONG X Y, HE X, LI X X, et al. The roles and functional mechanisms of interleukin-17 family cytokines in mucosal immunity[J]. Cell Mol Immunol, 2016, 13(4):418-431.
[16] LEE J S, TATO C M, JOYCE-SHAIKH B, et al. Interleukin-23-independent IL-17 production regulates intestinal epithelial permeability[J]. Immunity, 2015, 43(4):727-738.
[17] MAXWELL J R, ZHANG Y, BROWN W A, et al. Differential roles for interleukin-23 and interleukin-17 in intestinal immunoregulation[J]. Immunity, 2015, 43(4):739-750.
[18] IWAKURA Y, ISHIGAME H, SAIJO S, et al. Functional specialization of interleukin-17 family members[J]. Immunity, 2011, 34(2):149-162.
[19] RAMIREZ-CARROZZI V, SAMBANDAM A, LUIS E, et al.IL-17C regulates the innate immune function of epithelial cells in an autocrine manner[J]. Nat Immunol, 2011, 12(12):1159-1166.
[20] SONG X Y, ZHU S, SHI P Q, et al.IL-17RE is the functional receptor for IL-17C and mediates mucosal immunity to infection with intestinal pathogens[J]. Nat Immunol, 2011, 12(12):1151-1158.
[21] HUANG J L, MENG S H, HONG S J, et al.IL-17C is required for lethal inflammation during systemic fungal infection[J]. Cell Mol Immunol, 2016, 13(4):474-483.
[22] LOOS M, GEENS M, SCHAUVLIEGE S, et al. Role of heat-stable enterotoxins in the induction of early immune responses in piglets after infection with enterotoxigenic Escherichia coli[J]. PLoS One, 2012, 7(7):e41041.
[23] LUO Y, VAN NGUYEN U, DE LA FE RODRIGUEZ P Y, et al. F4⁺ ETEC infection and oral immunization with F4 fimbriae elicits an IL-17-dominated immune response[J]. Vet Res, 2015, 46:121.
[24] JIAO W, MA Q, LV X, et al. Gene expression and tissue distribution of β-defensins in Chinese Min pigs and Landrace pigs[J].Czech J Anim Sci, 2017, 62(4):178-183.
[25] PASTERNAK J A, AIYER V I A, HAMONIC G, et al. Molecular and physiological effects on the small intestine of weaner pigs following feeding with deoxynivalenol-contaminated feed[J]. Toxins (Basel), 2018, 10(1):40.
[26] BRUEL T, GUIBON R, MELO S, et al. Epithelial induction of porcine suppressor of cytokine signaling 2(SOCS2) gene expression in response to Entamoeba histolytica[J]. Dev Comp Immunol, 2010, 34(5):562-571.
[27] VON DER HARDT K, KANDLER M A, FINK L, et al. High frequency oscillatory ventilation suppresses inflammatory response in lung tissue andmicrodissected alveolar macrophages in surfactant depleted piglets[J]. Pediatr Res, 2004, 55(2):339-346.
[28] MELKEBEEK V, SONCK E, VERDONCK F, et al. Optimized FaeG expression and a thermolabile enterotoxin DNA adjuvant enhance priming of an intestinal immune response by an FaeG DNA vaccine in pigs[J]. Clin Vaccine Immunol, 2007, 14(1):28-35.
[29] STARNES T, BROXMEYER H E, ROBERTSON M J, et al. Cutting edge:IL-17D, a novel member of the IL-17 family, stimulates cytokine production and inhibits hemopoiesis[J]. J Immunol, 2002, 169(2):642-646.
[30] FRIEDRICH M, DIEGELMANN J, SCHAUBER J, et al. Intestinal neuroendocrine cells and goblet cells are mediators of IL-17A-amplified epithelial IL-17C production in human inflammatory bowel disease[J]. Mucosal Immunol, 2015, 8(4):943-958.
[31] IM E, JUNG J, RHEE S H. Toll-like receptor 5 engagement induces interleukin-17C expression in intestinal epithelial cells[J]. J Interf Cytok Res, 2012, 32(12):583-591.
[32] PFEIFER P, VOSS M, WONNENBERG B, et al.IL-17C is a mediator of respiratory epithelial innate immune response[J]. Am J Respir Cell Mol Biol, 2013, 48(4):415-421.
[33] KUSAGAYA H, FUJISAWA T, YAMANAKA K, et al. Toll-like receptor-mediated airway IL-17C enhances epithelial host defense in an autocrine/paracrine manner[J]. Am J Respir Cell Mol Biol, 2014, 50(1):30-39.
[34] REYNOLDS J M, ANGKASEKWINAI P, DONG C.IL-17 family member cytokines:regulation and function in innate immunity[J]. Cytok Growth Factor Rev, 2010, 21(6):413-423.
[35] ZAPH C, DU Y R, SAENZ S A, et al. Commensal-dependent expression of IL-25 regulates the IL-23-IL-17 axis in the intestine[J]. J Exp Med, 2008, 205(10):2191-2198.
[36] SAWA S, LOCHNER M, SATOH-TAKAYAMA N, et al.RORγt⁺ innate lymphoid cells regulate intestinal homeostasis by integrating negative signals from the symbiotic microbiota[J]. Nat Immunol, 2011, 12(4):320-326.

The Expression and Functional Analysis of IL-17 Cytokine Family in Enterotoxigenic E.coli Infected IPEC-J2 Cells

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	GAO Jie, LI Xiaocheng, MU Yang, ZHANG Hui, WEI Rong, LI Jie. Biological Characteristics and Immune Effect Evaluation of Outer Membrane Vesicles of Capsular Type B Pasteurella multocida [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2168-2175.
[2]	SU Wennan, LIU Jiaqi, ZHONG Jiacheng, CHEN Jidang, ZHU Wanjun, ZHANG Yishan, ZHANG Jipei. Complete Genome Re-sequence and Comparative Genomic Analysis of Avibacterium paragallinarum from Geese [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1208-1216.
[3]	ZHUANG Cuicui, HAN Bo. Mechanism of Mitochondrial Damage in Bovine Mammary Epithelial Cells and Mouse Mammary Gland Infected with Escherichia coli Isolated from Bovine Mastitis [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 822-833.
[4]	ZHI Yan, MEI Chen, LIU Zhenyi, WUYUN Gerile, WANG Hongjun, HU Ge. Research Progress on the Virulence Factors of Avibacterium paragallinarum [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 4934-4942.
[5]	WANG Jianing, ZHANG Ziqiang, KONG Dejing, FENG Caicai, ZHANG Feike, LIU Yumei. Isolation and Identification of Klebsiella pneumoniae in Rabbits [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5198-5206.
[6]	LI Lili, CHEN Kaifeng, CHEN Bing, ZHOU Zhouping, WANG Nanwei, QU Xiaoyun, XU Chenggang, LIAO Ming, ZHANG Jianmin. Regulatory Role of STM1827 in the Biofilm Formation and Environmental Stress of Salmonella Typhimurium [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5207-5217.
[7]	TONG Panpan, HUANG Shunmin, WANG Yudan, SHI Xuhui, CHEN Wenxia, SONG Xinlong, ZHANG Yi, SU Zhanqiang, XIE Jinxin. Phylogenetic Clustering, Serotype and Drug Resistance Analysis of Escherichia coli from Diarrhea with Piglets in Xinjiang [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 414-420.
[8]	WANG Fei, YANG Jie, Lü Qingjie, WANG Mixue, LIU Peng, ZHANG Ruoyu, SHI Congcong, WANG Xueying, LIN Lin, HUA Lin, SONG Wenbo, LIANG Wan, CHEN Huanchun, WU Bin, PENG Zhong. Isolation and Genomic Characterization of a Meningitis Causing Pasteurella multocida [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4346-4355.
[9]	MEN Kaikai, LIU Jialong, GUO Yage, HE Lei, JIA Yanyan, YU Zuhua. Effects of Chicken TGF-β1 on the Adhesion of Escherichia coli and Salmonella pullorum to DF1 Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 4000-4007.
[10]	YANG Yang, XIE Liqing, HU Pei, GAO Lixu, YUAN Xiang, LI Pan, PENG Yuanyi, LI Nengzhang. Construction of Bovine Pasteurella multocida Type A hyaD Mutant and Its Cross-protection Analysis in Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(10): 3582-3597.
[11]	SHAO Ying, FU Dandan, WU Xiaoyan, GU Yi, SONG Xiangjun, TU Jian, QI Kezong. Effect of the ETT2 Structural Gene epaPQR on the Biological Properties and Pathogenicity of Avian Pathogenic Escherichia coli [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(10): 3631-3641.
[12]	XU Wenbo, WU Limei, LIU Xin, LI Sheng, HUANG Fushen, LI Runcheng. Analysis on Complete Genome Sequence and Pathogenic Genes of a Pasteurella multocida Strain [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1858-1869.
[13]	WANG Xi, LI Ke, LI Tingcui, YAN Hongya, ZHAO Rong, CHANG Zhishun, LIAO Ming, SUN Minhua, XIN Aiguo. MLST Typing and Drug Resistance Analysis of 75 Salmonella Strains Isolated from Laying Hens [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(5): 1626-1631.
[14]	WANG Jun, LI Jun, CUI Guolin. The Effect of R91S Mutation in FliC on the Flagellar Shape and Salmonella Enteritidis Colonization in BALB/c Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 607-617.
[15]	TONG Panpan, ZHANG Mengmeng, CHEN Wenxia, LIU Luyao, ZHANG Ling, TANG Xuelin, SU Zhanqiang, XIE Jinxin. Phylogenetic Clustering, Serotype, Virulence Genes and Drug Resistance Analysis of Shiga Toxin-producing Escherichia coli from Cattle, Sheep and Camel in Xinjiang [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3660-3668.