IL-17抑制铜绿假单胞菌感染肺上皮细胞的机制研究

doi:10.19586/j.2095-2341.2023.0151

生物技术进展 ›› 2024, Vol. 14 ›› Issue (2): 304-311.DOI: 10.19586/j.2095-2341.2023.0151

• 研究论文 • 上一篇

IL-17抑制铜绿假单胞菌感染肺上皮细胞的机制研究

王彩虹(), 邵煜涵, 蒋梦媛

首都医科大学附属北京友谊医院，北京热带医学研究所，北京 100050

收稿日期:2023-11-23 接受日期:2023-12-11 出版日期:2024-03-25 发布日期:2024-04-17
作者简介:王彩虹 E-mail：13520137104@163.com
基金资助:
北京市医管中心青苗项目(QML20230105)

The Mechanisms of IL-17 Inhibiting Pseudomonas aeruginosa Infection on Lung Epithelial Cells

Caihong WANG(), Yuhan SHAO, Mengyuan JIANG

Beijing Institute of Tropical Medicine，Beijing Friendship Hospital，Capital Medical University，Beijing 100050，China

Received:2023-11-23 Accepted:2023-12-11 Online:2024-03-25 Published:2024-04-17

摘要/Abstract

摘要：

铜绿假单胞菌是常见的致病菌，白细胞介素-17(interleukin-17，IL-17)家族细胞因子直接参与宿主免疫应答，是宿主防御感染或炎症的关键介质。利用重组人IL-17蛋白处理铜绿假单胞菌PAO1，探索其对肺上皮细胞A549的侵袭作用。结果发现，在不影响细菌生长和细胞增殖的情况下，IL-17能够抑制PAO1对A549细胞的入侵。利用转录组测序分析IL-17对PAO1转录组的改变，探索IL-17影响PAO1入侵宿主细胞的作用机制。结果表明，有210个差异基因表达上调，123个表达下调；GO功能分析显示差异基因主要富集在肽酶活性、蛋白结合和蛋白水解等方面；KEGG富集分析显示，差异基因涉及RNA降解、生物膜形成、微生物代谢、ABC转运以及碳代谢等通路。此外，生物膜形成实验进一步证实了IL-17能够降低PAO1生物膜的形成。研究结果为耐药铜绿假单胞菌在临床感染造成的相关疾病的治疗提供了新思路和理论基础。

关键词: 铜绿假单胞菌, 感染, 转录组, 白细胞介素-17, 肺上皮细胞

Abstract:

Pseudomonas aeruginosa is a common pathogen. Interleukin-17 （IL-17） family cytokines are directly involved in the host immune response， which are key mediators of the defense against infection or inflammation in host. Using recombinant human IL-17 protein to treat Pseudomonas aeruginosa PAO1 and explore its invasive effect on lung epithelial cell A549， it was found that IL-17 can inhibit the invasion of PAO1 to A549 cells without affecting bacterial growth and cell proliferation. To explore the mechanism of IL-17 influencing PAO1 infection on host cells， we utilized transcriptome sequencing to analyze the changes of IL-17 on the PAO1 transcriptome. The results showed that there are 210 upregulated genes and 123 downregulated. GO functional analysis showed that the differential genes were mainly enriched in peptidase activity， protein binding and proteolysis. KEGG enrichment analysis showed that the differential genes were involved in RNA degradation， biofilm formation， microbial metabolism， ABC transport and carbon metabolism. In addition， the biofilm formation experiments further confirmed that IL-17 could reduce PAO1 biofilm formation. These findings provided a new idea and theoretical basis for the treatment of drug-resistant Pseudomonas aeruginosa in the treatment of related diseases caused by clinical infection.

Key words: Pseudomonas aeruginosa, infection, transcriptome, interleukin-17, lung epithelial cell

中图分类号:

Q291

王彩虹, 邵煜涵, 蒋梦媛. IL-17抑制铜绿假单胞菌感染肺上皮细胞的机制研究[J]. 生物技术进展, 2024, 14(2): 304-311.

Caihong WANG, Yuhan SHAO, Mengyuan JIANG. The Mechanisms of IL-17 Inhibiting Pseudomonas aeruginosa Infection on Lung Epithelial Cells[J]. Current Biotechnology, 2024, 14(2): 304-311.

图/表 8

图1 不同浓度IL-17对A549细胞生长的影响注：***表示两组数据间在P<0.001水平有统计学差异，ns表示无统计学差异。

Fig. 1 The effect of different concentrations of IL-17 on the growth of A549 cells

图2 不同浓度IL-17对PAO1生长的影响

Fig. 2 The effect of different concentrations of IL-17 on the growth of PAO1

图3 IL-17抑制PAO1入侵A549细胞注：**表示两组数据在P<0.01水平有统计学差异。

Fig. 3 IL-17 inhibits PAO1 invasion into A549 cells

图4 差异基因分布火山图注：图中横坐标表示基因在处理和对照两组中的表达倍数变化(log2FoldChange)，纵坐标表示基因在处理和对照两组中表达差异的显著性水平(-lgPadj)。

Fig. 4 Volcano map of differential gene distribution

图5 GO功能分类图

Fig. 5 GO function classification diagram

图6 GO富集结果图

Fig. 6 GO enrichment result chart

图7 KEGG富集分析结果

Fig. 7 KEGG enrichment analysis results

图8 IL-17影响生物膜形成注：***表示在P<0.001水平具有统计学意义。

Fig. 8 IL-17 affects biofilm formation

参考文献 29

1	QIN S, XIAO W, ZHOU C, et al.. Pseudomonas aeruginosa: pathogenesis, virulence factors, antibiotic resistance, interaction with host, technology advances and emerging therapeutics[J/OL]. Signal. Transduct. Target. Ther., 2022, 7(1): 199[2023-12-02]. .
2	曾桃花,李文茹,谢小保,等.植物精油对铜绿假单胞菌群体感应系统的抑制作用研究进展[J].生物技术进展,2021,11(3):279-288.
	ZENG T H, LI W R, XIE X B, et al.. Research progress in the inhibitory effects of essential oils on the quorum sensing system of Pseudomonas aeruginosa [J]. Curr. Biotechnol., 2021, 11(3): 279-288.
3	CURRAN C S, BOLIG T, TORABI-PARIZI P. Mechanisms and targeted therapies for Pseudomonas aeruginosa lung infection[J]. Am. J. Respir. Crit. Care Med., 2018, 197(6): 708-727.
4	MIDDLETON M A, LAYEGHIFARD M, KLINGEL M, et al.. Epidemiology of clonal Pseudomonas aeruginosa infection in a Canadian cystic fibrosis population[J]. Ann. Am. Thorac. Soc., 2018, 15(7): 827-836.
5	OCHMAN H, MORAN N A. Genes lost and genes found: evolution of bacterial pathogenesis and symbiosis[J]. Science, 2001, 292(5519): 1096-1099.
6	ANTUNES M B, COHEN N A. Mucociliary clearance: a critical upper airway host defense mechanism and methods of assessment[J]. Curr. Opin. Allergy Clin. Immunol., 2007, 7(1): 5-10.
7	O'CONNELL K M, HODGKINSON J T, SORE H F, et al.. Combating multidrug-resistant bacteria: current strategies for the discovery of novel antibacterials[J]. Angew. Chem. Int. Ed., 2013, 52(41): 10706-10733.
8	MCGEACHY M J, CUA D J, GAFFEN S L. The IL-17 family of cytokines in health and disease[J]. Immunity, 2019, 50(4): 892-906.
9	PAPPU R, RUTZ S, OUYANG W. Regulation of epithelial immunity by IL-17 family cytokines[J]. Trends Immunol., 2012, 33(7): 343-349.
10	IWAKURA Y, ISHIGAME H, SAIJO S, et al.. Functional specialization of interleukin-17 family members[J]. Immunity, 2011, 34(2): 149-162.
11	ASADA Y, SUZUKI I, SUZUKI M, et al.. Atypical multiple benign osteoblastomas accompanied by simple bone cysts: a case report[J]. J. Cranio Maxill. Surg., 1991, 19(4): 166-171.
12	JEON Y J, JO A, WON J, et al.. IL-17C protects nasal epithelium from Pseudomonas aeruginosa infection[J]. Am. J. Respir. Cell Mol. Biol., 2020, 62(1): 95-103.
13	PAPAZIAN L, KLOMPAS M, LUYT C E. Ventilator-associated pneumonia in adults: a narrative review[J]. Intensive Care Med., 2020, 46(5): 888-906.
14	PALAVUTITOTAI N, JITMUANG A, TONGSAI S, et al.. Epidemiology and risk factors of extensively drug-resistant Pseudomonas aeruginosa infections[J/OL]. PLoS ONE, 2018, 13(2): e0193431[2018-02-23]. .
15	RICHARDS M J, EDWARDS J R, CULVER D H, et al.. Nosocomial infections in combined medical-surgical intensive care units in the United States[J]. Infect. Contr. Hosp. Epidemiol., 2000, 21(8): 510-515.
16	LISTER P D, WOLTER D J, HANSON N D. Antibacterial-resistant Pseudomonas aeruginosa: clinical impact and complex regulation of chromosomally encoded resistance mechanisms[J]. Clin. Microbiol. Rev., 2009, 22(4): 582-610.
17	GILADI M, PORAT Y, BLATT A, et al.. Microbial growth inhibition by alternating electric fields in mice with Pseudomonas aeruginosa lung infection[J]. Antimicrob. Agents Chemother., 2010, 54(8): 3212-3218.
18	NOORANIDOOST M, COGAN N G, STOODLEY P, et al.. Bayesian estimation of Pseudomonas aeruginosa viscoelastic properties based on creep responses of wild type, rugose, and mucoid variant biofilms[J/OL]. Biofilm, 2023, 5:100133[2023-07-03]. .
19	GAGNE-THIVIERGE C, BARBEAU J, LEVESQUE R C, et al.. A new approach to study attached biofilms and floating communities from Pseudomonas aeruginosa strains of various origins reveals diverse effects of divalent ions[J/OL]. FEMS Microbiol. Lett., 2018, 365: 14[2018-06-27]. .
20	DAVIES J, DAVIES D. Origins and evolution of antibiotic resistance[J]. Microbiol. Mol. Biol. Rev., 2010, 74(3): 417-433.
21	LE ROUZIC O, PICHAVANT M, FREALLE E, et al.. Th17 cytokines: novel potential therapeutic targets for COPD pathogenesis and exacerbations[J]. Eur. Respir. J., 2017, 50: 4[2023-12-02]. .
22	CHEN K, POCIASK D A, MCALEER J P, et al.. IL-17RA is required for CCL2 expression, macrophage recruitment, and emphysema in response to cigarette smoke[J/OL]. PLoS ONE, 2011, 6(5): e20333[2023-01-02]. .
23	XU X, SHAO B, WANG R, et al.. Role of interleukin-17 in defense against pseudomonas aeruginosa infection in lungs[J]. Int. J. Clin. Exp. Med., 2014, 7(4): 809-816.
24	CONTI H R, GAFFEN S L. IL-17-mediated immunity to the opportunistic fungal pathogen Candida albicans [J]. J. Immunol. 2015, 195(3): 780-788.
25	HAPPEL K I, ZHENG M, YOUNG E, et al.. Cutting edge: roles of Toll-like receptor 4 and IL-23 in IL-17 expression in response to Klebsiella pneumoniae infection[J]. J. Immunol., 2003, 170(9): 4432-4436.
26	MA W T, YAO X T, PENG Q, et al.. The protective and pathogenic roles of IL-17 in viral infections: friend or foe?[J/OL] Open Biol., 2019, 9(7): 190109[2019-07-25]. .
27	BAYES H K, RITCHIE N D, EVANS T J. Interleukin-17 is required for control of chronic lung infection caused by Pseudomonas aeruginosa [J]. Infect. Immun., 2016, 84(12): 3507-3516.
28	MEYER N J, GATTINONI L, CALFEE C S. Acute respiratory distress syndrome[J]. Lancet Lond. Engl., 2021, 398(10300): 622-637.
29	ME R, GAO N, DAI C, et al.. IL-17 promotes Pseudomonas aeruginosa keratitis in C57BL/6 mouse corneas[J]. J. Immunol., 2020, 204(1): 169-179.

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IL-17抑制铜绿假单胞菌感染肺上皮细胞的机制研究

The Mechanisms of IL-17 Inhibiting Pseudomonas aeruginosa Infection on Lung Epithelial Cells

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