LIPI-3和LIPI-4共存对致病性单核细胞增生李斯特菌毒力的影响

doi:10.16431/j.cnki.1671-7236.2024.05.024

Abstract

Abstract: 【Objective】 This study was aimed to explore the impact of the coexistence of LIPI-3 and LIPI-4 on the virulence of Listeria monocytogenes (LM),reveal their potential regulatory effects on bacterial virulence,and lay a foundation for further research on the pathogenic mechanisms of LM.【Method】 LM928 (possessing LIPI-4 present) and LM873 (possessing both LIPI-3 and LIPI-4) were used as research objects to monitor the adhesion and invasion of two strains by infecting HCMEC/D3 cells.The effect of two strains on the median lethal dose (LD₅₀) of chicken embryos were evaluated by chicken embryo infection test.The hemolytic activity of two strains were determined using hemolysis test.The apoptosis of HCMEC/D3 cells was analyzed using flow cytometry to evaluate the ability of two strain to induce cell apoptosis.The transcriptional differences of virulence genes between two strains under BHI culture conditions and post-infection of HCMEC/D3 cells were compared using Real-time quantitative PCR.【Result】 There was no significant difference of adhesion rates between LM928 and LM873 strains (P＞0.05).The invasion rate of LM928 strain was extremely significantly higher than that of LM873 strain (P＜0.01).The LD₅₀ of LM873 strain was approximately 1 000 times that of LM928 strain.The hemolytic values of LM928 and LM873 strains were 2⁴ and 2³,respectively.The apoptosis rate induced by LM928 strain at 24 and 48 h post-infection were significantly or extremely significantly higher than that of LM873 strain (P＜0.05 or P＜0.01),while there was no significant difference of the apoptosis rate between LM873 strain and control group (P＞0.05).When cultured in BHI culture,compared with LM928 strain,the transcription levels of virulence genes mpl,inlB,inlC,inlP,actA,plcA,plcB and sigB in LM873 strain were significantly or extremely significantly upregulated (P＜0.05 or P＜0.01),and the transcription levels of hly,inlA and iap genes were extremely significantly or significantly downregulated (P＜0.01 or P＜0.05).After infecting HCMEC/D3 cells,compared with LM928 strain,the transcription levels of virulence genes inlP,actA,plcA,plcB and prfA in LM873 strain were significantly or extremely significantly upregulated (P＜0.05 or P＜0.01),and the transcription levels of hly,mpl,inlA,inlB,inlC and iap genes were extremely significantly downregulated (P＜0.01).【Conclusion】 The coexistence of LIPI-3 and LIPI-4 reduced the virulence of LM,which holded significant implications for further research into the molecular mechanisms and complex interactions of LIPI-3 and LIPI-4 in bacterial virulence regulation.

Key words: Listeria monocytogenes (LM); LIPI-3; LIPI-4; virulence; virulence gene

CLC Number:

S852.61⁺7

QIAN Ruixuan, LI Nan, KANG Lichao, MA Xun, LIU Caixia, SHI Weidi, KOU Lijun, REN Huijie, QI Yatao, YIN Zhongke, LIU Lu, WANG Jing, WANG Zhengrong, JIANG Jianjun. Influence of Coexistence of LIPI-3 and LIPI-4 on Virulence of Pathogenic Listeria monocytogenes[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 2027-2036.

References

[1] SWAMINATHAN B,GERNER-SMIDT P.The epidemiology of human listeriosis[J].Microbes and Infection,2007,9(10):1236-1243.
[2] VÁZQUEZ-BOLAND J A,KUHN M,BERCHE P,et al.Listeria pathogenesis and molecular virulence determinants[J].Clinical Microbiology Reviews,2001,14(3):584-640.
[3] MAURY M M,TSAI Y H,CHARLIER C,et al.Uncovering Listeria monocytogenes hypervirulence by harnessing its biodiversity[J].Nature Genetics,2016,48(3):308-313.
[4] 李红欢,陈朔,康立超,等.食源性单增李斯特菌毒力岛基因检测与致病性[J].江苏农业科学,2019,47(14):193-196. LI H H,CHEN S,KANG L C,et al.Pathogenicity island gene detection and pathogenicity of food-borne Listeria monocytogenes[J].Jiangsu Agricultural Sciences,2019,47(14):193-196.(in Chinese)
[5] LEE S.Bacteriocins of Listeria monocytogenes and their potential as a virulence factor[J]. Toxins,2020,12(2):103.
[6] 刘扬扬,康立超,马勋,等.食源性单增李斯特菌LIPI-4基因的检测及序列分析[J].中国兽医学报,2019,39(12):2380-2385. LIU Y Y,KANG L C,MA X,et al.Detection and sequence analysis of LIPI-4 gene on food-borne Listeria monocytogenes[J]. Chinese Journal of Veterinary Science,2019,39(12):2380-2385.(in Chinese)
[7] 刘彩霞,高盛杰,寇丽君,等.LIPI-4 EⅡA基因对单核细胞增生性李斯特菌重要毒力基因转录调控作用的研究[J].中国预防兽医学报,2022,44(12):1269-1276. LIU C X,GAO S J,KOU L J,et al.Transcriptional regulation of important virulence genes in Listeria monocytogenes by permease ⅡA of Listeria pathogenicity island 4[J].Chinese Journal of Preventive Veterinary Medicine,2022,44(12):1269-1276.(in Chinese)
[8] 康立超,钱晶,杜冬冬,等.新疆食源性单增李斯特菌MLST分型及耐药性分析[J].食品安全质量检测学报,2021,12(15):6255-6261. KANG L C,QIAN J,DU D D,et al.Multilocus sequence typing and antibiotic resistance of food-borne Listeria monocytogenes in Xinjiang[J].Journal of Food Safety & Quality,2021,12(15):6255-6261.(in Chinese)
[9] 张琴.肉鸡屠宰与销售环节单核细胞增生李斯特菌的分子流行特征及有机酸减菌技术的研究[D].扬州:扬州大学,2023. ZHANG Q.Molecular epidemiological characteristics of Listeria monocytogenes in the chicken slaughtering and sale process and decontamination of Listeria technology using organic acid[D].Yangzhou:Yangzhou University,2023.(in Chinese)
[10] 炊慧霞,李薇薇,崔箐坡,等.河南省2015至2019年李斯特菌病分子流行病学特征[J].郑州大学学报(医学版),2021,56(6):767-773. CHUI H X,LI W W,CUI Q P,et al.Molecular epidemiological characteristics of listeriosis in Henan from 2015 to 2019[J]. Journal of Zhengzhou University(Medical Sciences),2021,56(6):767-773.(in Chinese)
[11] QUEREDA J J,MORÓN-GARCÍA A,PALACIOS-GORBA C,et al.Pathogenicity and virulence of Listeria monocytogenes:A trip from environmental to medical microbiology[J].Virulence,2021,12(1):2509-2545.
[12] JOHNSON L J,AZARI S,WEBB A,et al.Human placental trophoblasts infected by Listeria monocytogenes undergo a pro-inflammatory switch associated with poor pregnancy outcomes[J].Frontiers in Immunology,2021,12:709466.
[13] LI W,GUO Y,CUI Q,et al.Whole-genome sequencing-based characterization of clinical Listeria monocytogenes isolates in China,2013-2019[J]. Foodborne Pathogens and Disease,2023,20(4):158-168.
[14] MYINTZAW P,PENNONE V,MCAULIFFE O,et al.Association of virulence,biofilm,and antimicrobial resistance genes with specific clonal complex types of Listeria monocytogenes[J]. Microorganisms,2023,11(6):1603.
[15] CENTOROTOLA G,ZIBA M W,CORNACCHIA A,et al.Listeria monocytogenes in ready to eat meat products from Zambia:Phenotypical and genomic characterization of isolates[J].Frontiers in Microbiology,2023,14:1228726.
[16] COTTER P D,DRAPER L A,LAWTON E M,et al.Listeriolysin S,a novel peptide haemolysin associated with a subset of lineage Ⅰ Listeria monocytogenes[J]. PLoS Pathogens,2008,4(9):e1000144.
[17] WIKTORCZYK-KAPISCHKE N,SKOWRON K,WAECKA-ZACHARSKA E.Genomic and pathogenicity islands of Listeria monocytogenes-Overview of selected aspects[J]. Frontiers in Molecular Biosciences,2023,10:1161486.
[18] 刘扬扬,康立超,马勋,等.食源性单核细胞增生李斯特菌毒力岛4携带株的分型、毒力及生物被膜形成能力的测定[J].中国预防兽医学报,2020,42(6):567-571. LIU Y Y,KANG L C,MA X,et al.Typing,virulence and biofilm formation of the food-borne Listeria monocytogenes carrying pathogenicity island 4[J]. Chinese Journal of Preventive Veterinary Medicine,2020,42(6):567-571.(in Chinese)
[19] LEE S,PARSONS C,CHEN Y,et al.Contrasting genetic diversity of Listeria pathogenicity islands 3 and 4 harbored by nonpathogenic Listeria spp.[J]. Applied and Environmental Microbiology,2023,89(2):e02097-22.
[20] 刘武康,陈国薇,谢曼曼,等.毒力因子InlA和InlB缺失影响单增李斯特菌侵袭宿主细胞和诱导凋亡的能力[J].食品科学,2017,38(19):49-54. LIU W K,CHEN G W,XIE M M,et al.Effect of deletion of genes encoding virulence factors InlA and InlB on the ability of Listeria monocytogenes to invade host cells and induce cell apoptosis[J]. Food Science,2017,38(19):49-54.(in Chinese)
[21] PHELPS C C,VADIA S,ARNETT E,et al.Relative roles of listeriolysin O,InlA,and InlB in Listeria monocytogenes uptake by host cells[J].Infection and Immunity,2018,86(10):e00555-18.
[22] KÖHLER S,LEIMEISTER-WÄCHTER M,CHAKRABORTY T,et al.The gene coding for protein p60 of Listeria monocytogenes and its use as a specific probe for Listeria monocytogenes[J].Infection and Immunity,1990,58(6):1943-1950.
[23] 王政,张闻桐,吴美娇,等.单增李斯特菌溶血素O的功能研究进展[J].微生物学杂志,2020,40(4):98-104. WANG Z,ZHANG W T,WU M J,et al.Advances in function of Listeria monocytogenes listeriolysin O (LLO)[J].Journal of Microbiology,2020,40(4):98-104.(in Chinese)
[24] CARRERO J A,UNANUE E R.Mechanisms and immunological effects of apoptosis caused by Listeria monocytogenes[J].Advances in Immunology,2012,113:157-174.
[25] ZHANG T,BAE D,WANG C.Listeriolysin O mediates cytotoxicity against human brain microvascular endothelial cells[J].FEMS Microbiology Letters,2015,362(12):fnv084.
[26] NAVARRE W W,PORWOLLIK S,WANG Y,et al.Selective silencing of foreign DNA with low GC content by the H-NS protein in Salmonella[J].Science (New York,N.Y.),2006,313(5784):236-238.
[27] SIRYAPORN A,GOULIAN M.Cross-talk suppression between the CpxA-CpxR and EnvZ-OmpR two-component systems in E.coli[J]. Molecular Microbiology,2008,70(2):494-506.
[28] KWOK R.Five hard truths for synthetic biology[J]. Nature,2010,463(7279):288-290.
[29] SAITO H.Regulation of cross-talk in yeast MAPK signaling pathways[J].Current Opinion in Microbiology,2010,13(6):677-683.
[30] MVLLER I E,RUBENS J R,JUN T,et al.Gene networks that compensate for crosstalk with crosstalk[J].Nature Communications,2019,10:4028.
[31] ALONSO-ROMAN R,LAST A,MIRHAKKAK M H,et al.Lactobacillus rhamnosus colonisation antagonizes Candida albicans by forcing metabolic adaptations that compromise pathogenicity[J].Nature Communications,2022,13(1):3192.

Influence of Coexistence of LIPI-3 and LIPI-4 on Virulence of Pathogenic Listeria monocytogenes

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