金黄色葡菌球菌脂蛋白对M1型小鼠骨髓源巨噬细胞免疫作用的影响

doi:10.16431/j.cnki.1671-7236.2021.08.041

中国畜牧兽医 ›› 2021, Vol. 48 ›› Issue (8): 3079-3086.doi: 10.16431/j.cnki.1671-7236.2021.08.041

金黄色葡菌球菌脂蛋白对M1型小鼠骨髓源巨噬细胞免疫作用的影响

张静^1,2, 钱英红³, 张凯^1,2, 吴金迪^1,2, 刘博^1,2, 毛伟^1,2, 曹金山^1,2

1. 内蒙古农业大学兽医学院, 呼和浩特 010018;
2. 农业农村部动物疾病临床诊疗技术重点实验室, 呼和浩特 010018;
3. 内蒙古自治区农牧业科学院, 呼和浩特 010018

收稿日期:2021-03-05 出版日期:2021-08-20 发布日期:2021-08-19
通讯作者: 毛伟, 曹金山 E-mail:maowei820115@163.com;jinshancao@imau.edu.cn
作者简介:张静(1993-),女,内蒙古赤峰人,硕士生,研究方向:临床兽医学,E-mail:240206605@qq.com
基金资助:
国家自然科学基金（31860720）；内蒙古自治区科学计划（2020GG0042）；内蒙古自治区自然科学基金（2019MS03053）

Effects of Staphylococcus aureus Lipoprotein on Immunity in M1 Mouse Bone Marrow-Derived Macrophages

ZHANG Jing^1,2, QIAN Yinghong³, ZHANG Kai^1,2, WU Jindi^1,2, LIU Bo^1,2, MAO Wei^1,2, CAO Jinshan^1,2

1. College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. Key Laboratory of Animal Disease Clinical Diagnosis, Ministry of Agriculture and Rural Affairs, Hohhot 010018, China;
3. Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Hohhot 010018, China

Received:2021-03-05 Online:2021-08-20 Published:2021-08-19

摘要/Abstract

摘要： 本研究旨在阐明金黄色葡萄球菌脂蛋白对M1型小鼠骨髓源巨噬细胞免疫作用的影响，为金黄色葡萄球菌致病性研究提供理论参考。以金黄色葡萄球菌野生株SA113（WT SA113）和SA113 lgt：：ermB脂蛋白表达缺失菌株（SA113Δlgt株）体外感染M1型小鼠骨髓源巨噬细胞，分为3组：空白对照组、WT SA113感染组（MOI：3：1）、SA113Δlgt感染组（MOI：3：1）。采用ELISA法检测M1型小鼠骨髓源巨噬细胞中肿瘤坏死因子-α（TNF-α）、白细胞介素1β（IL-1β）、趋化因子（RANTES）和白细胞介素10（IL-10）的水平，实时荧光定量PCR检测Toll样受体2（TLR2）、Toll样受体4（TLR4）和含NLR家族Pyrin域蛋白3（NLRP3）基因的表达，免疫荧光法检测脂蛋白对M1型小鼠骨髓源巨噬细胞吞噬金黄色葡萄球菌作用的影响。结果显示，与空白对照组相比，WT SA113感染组和SA113Δlgt感染组均可显著上调M1型小鼠骨髓源巨噬细胞中TNF-α、RANTES、IL-10分泌量以及WT SA113感染组TLR2、NLRP3基因表达水平（P<0.05），而TLR4基因表达量显著降低（P<0.05）；与WT SA113感染组相比，SA113Δlgt感染组M1型小鼠骨髓源巨噬细胞中TNF-α、IL-1β、RANTES、IL-10分泌量以及TLR2（12 h除外）、NLRP3基因表达水平显著降低（P<0.05）。免疫荧光结果显示，M1型巨噬细胞对SA113Δlgt株的吞噬作用显著低于对WT SA113株的吞噬作用（P<0.05）。综上，金黄色葡萄球菌的脂蛋白在M1型小鼠骨髓源巨噬细胞中主要通过激活TLR2和NLRP3受体，诱导细胞因子TNF-α、IL-1β、RANTES和IL-10的产生和释放。

关键词: 金黄色葡萄球菌; 脂蛋白; 骨髓源巨噬细胞; 细胞因子

Abstract: The aim of this study was to clarify the effect of Staphylococcus aureus lipoproteins on immunity of M1 mouse bone marrow-derived macrophages, and provide the theoretical reference for the study of Staphylococcus aureus pathogenicity.In vitro, M1 mouse bone marrow-derived macrophages was infected by WT SA113 and SA113 lgt::ermB strains (SA113Δlgt) of Staphylococcus aureus.The mice were divided into three groups:Blank control group, WT SA113 infection group (MOI:3:1), and SA113Δlgt infection group (MOI:3:1).The tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), chemokines (RANTES) and interleukin-10 (IL-10) in M1 mouse bone marrow-derived macrophages was detected by ELISA method, the expression of Toll-like receptor 2 (TLR2), Toll-like receptor 2(TLR4) and recombinant NLR family, Pyrin domain containing protein 3 (NLRP3) genes were detected by Real-time quantitative PCR, and the effect of lipoproteins on phagocytosis of Staphylococcus aureus by M1 mouse bone marrow-derived macrophages was detected by immunofluorescence.The results showed that compared with control group, both WT SA113 and SA113Δlgt infection could significantly upregulate the secretion of TNF-α, RANTES and IL-10, and WT SA113 infection could significantly increase the expression of TLR2 and NLRP3 genes in M1 mouse bone marrow-derived macrophages polarization (P<0.05), while the expression of TLR4 gene was significantly decreased (P<0.05).Compared with WT SA113 infected group, the secretion of TNF-α, IL-1β, RANTES and IL-10, and the expression of TLR2 (expect for 12 h) and NLRP3 gene in SA113Δlgt infected group were significantly decreased (P<0.05).The immunofluorescence results showed that the phagocytosis effect of M1 macrophage on SA113Δlgt strain was significantly lower than that of WT SA113 strain (P<0.05).In conclusion, the lipoprotein of Staphylococcus aureus induced the production and release of cytokines TNF-α, IL-1β, RANTES and IL-10 mainly through activation of TLR2 and NLRP3 receptors in M1 mouse bone marrow-derived macrophages.

Key words: Staphylococcus aureus; lipoprotein; bone marrow-derived macrophages; cytokines

中图分类号:

S852.61⁺1

张静, 钱英红, 张凯, 吴金迪, 刘博, 毛伟, 曹金山. 金黄色葡菌球菌脂蛋白对M1型小鼠骨髓源巨噬细胞免疫作用的影响[J]. 中国畜牧兽医, 2021, 48(8): 3079-3086.

ZHANG Jing, QIAN Yinghong, ZHANG Kai, WU Jindi, LIU Bo, MAO Wei, CAO Jinshan. Effects of Staphylococcus aureus Lipoprotein on Immunity in M1 Mouse Bone Marrow-Derived Macrophages[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 3079-3086.

参考文献

[1] ARCHER G L.Staphylococcus aureus:A well-armed pathogen[J].Reviews of Infectious Diseases,1998,26(5):1179-1181.
[2] KLUYTMANS J,VAN BELKUM A,VERBRUGH H.Nasal carriage of Staphylococcus aureus:Epidemiology,underlying mechanisms,and associated risks[J].Clinical Microbiology Reviews,1997,10(3):505-520.
[3] BONE R C.Gram-positive organisms and sepsis[J].Archives of Internal Medicine,1994,154(1):26-34.
[4] FLUIT A C,SCHMITZ F J,VERHOEF J,et al.Frequency of isolation of pathogens from bloodstream,nosocomial pneumonia,skin and soft tissue,and urinary tract infections occurring in European patients[J].International Journal of Infectious Diseases,2001,20(3):188-191.
[5] PETIT T,SPERGSER J,ROSENGARTEN R,et al.Prevalence of potentially pathogenic bacteria as genital pathogens in dairy cattle[J].Reproduction in Domestic Animals,2009,44(1):88-91.
[6] 刘昆.PGE₂在金黄色葡萄球菌毒力因子诱导奶牛子宫内膜组织炎症因子表达过程中的调控作用及其机制研究[D].呼和浩特:内蒙古农业大学,2020. LIU K.Study on effect and mechanism of PGE₂ in process of inflammatory factors expression of bovine endometrial tissue induce by Staphylococcus aureus[D].Hohhot:Inner Mongolia Agricultural University,2020.(in Chinese)
[7] KRETSCHMER D,HANZELMANN D,PESCHEL A.Lipoprotein immunoproteomics question the potential of Staphylococcus aureus TLR2 agonists as vaccine antigens[J].Proteomics,2016,16(20):2603-2604.
[8] WANG C,CHEN T,ZHANG J,et al.The E3 ubiquitin ligase Nrdp1 ‘preferentially’ promotes TLR-mediated production of type interferon[J].Nature Immunology,2009,10(7):744-752.
[9] 吴金迪.金黄色葡萄球菌感染小鼠腹腔巨噬细胞的分子机制及前列腺素E₂对其影响的研究[D].呼和浩特:内蒙古农业大学,2018. WU J D.Molecular mechanism and the effect of prostaglandin E₂ regulate activation of mouse peritoneal macrophages by Staphylococcus aureus[D].Hohhot:Inner Mongolia Agricultural University,2018.(in Chinese)
[10] BENOIT M,DESNUES B,MEGE J.Macrophage polarization in bacterial infections[J].Journal of Immunology,2008,181(6):3733-3739.
[11] MōGE J L,MEHRAJ V,CAPO C.Macrophage polarization and bacterial infections[J].Current Opinion in Infectious Diseases,2011,24(3):230-234.
[12] NGUYEN M T,KRAFT B,YU W,et al.The νSaα specific lipoprotein like cluster (LPL) of S.aureus USA300 contributes to immune stimulation and invasion in human cells[J].PLoS Pathogens,2015,11(6):e1004984.
[13] TAWARATSUMIDA K,FURUYASHIKI M,KATSUMOTO M,et al.Characterization of N-terminal structure of TLR2-activating lipoprotein in Staphylococcus aureus[J].The Journal of Biological Chemistry,2009,284(14):9147-9152.
[14] STOLL H,DENGJEL J,NERZ C,et al.Staphylococcus aureus deficient in lipidation of prelipoproteins is attenuated in growth and immune activation[J].Infection and Immunity,2005,73(4):2411-2423.
[15] THIRIOT J D,MARTINEZ-MARTINEZ Y B,ENDSLEY J J,et al.Hacking the host:Exploitation of macrophage polarization by intracellular bacterial pathogens[J].Pathogens and Disease,2020,78(1):ftaa009.
[16] KENGMO T A,PESCHEL A.Staphylococcus aureus releases proinflammatory membrane vesicles to resist antimicrobial fatty acids[J].mSphere,2020,5(5):e00804-20.
[17] MOHAMMAD M,HU Z,ALI A,et al.The role of Staphylococcus aureus lipoproteins in hematogenous septic arthritis[J].Scientific Reports,2020,10(1):7936.
[18] BUWITT B U,HEINE H,WIESMVLLER K H,et al.TLR1-and TLR6-independent recognition of bacterial lipopeptides[J].The Journal of Biological Chemistry,2006,281:9049-9057.
[19] SCHMALER M,JANN NJ,FERRACIN F,et al.Lipoproteins in Staphylococcus aureus mediate inflammation by TLR2 and iron-dependent growth in vivo[J].Journal of Immunology,2009,182:7110-7118.
[20] WU J,LIU B,MAO W,et al.Prostaglandin E₂ regulates activation of mouse peritoneal macrophages by Staphylococcus aureus through Toll-like receptor 2,Toll-like receptor 4,and NLRP3 inflammasome signaling[J].Journal of Innate Immunity,2020,12(2):154-169.
[21] HASHIMOTO M,TAWARATSUMIDA K,KARIYA H,et al.Lipoprotein is a predominant Toll-like receptor 2 ligand in Staphylococcus aureus cell wall components[J].International Immunology,2006,18(2):355-362.
[22] FOWLER T,WANN E R,JOH D,et al.Cellular invasion by Staphylococcus aureus involves a fibronectin bridge between the bacterial fibronectin binding MSCRAMMs and host cell beta1 integrins[J].European Journal of Cell Biology,2000,79:672-679.

金黄色葡菌球菌脂蛋白对M1型小鼠骨髓源巨噬细胞免疫作用的影响

Effects of Staphylococcus aureus Lipoprotein on Immunity in M1 Mouse Bone Marrow-Derived Macrophages

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