Comparison in the characterization of outer membrane vesicles from Pseudomonas aeruginosa in normal culture and under meropenem pressure

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Journal of Microbes and Infections ›› 2024, Vol. 19 ›› Issue (3) : 129-139.

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Abstract

This aim of this study was to distinct the physical characterization and protein components of outer membrane vesicle (OMV) secreted from Pseudomonas aeruginosa under normal culture condition and meropenem (MEM) stimulated culture by a variety of technological means including transmission electron microscopy (TEM), Liquid chromatography tandem mass spectrometry (LC-MS/MS). At the same time, mouse macrophage cell line Raw264.7 was cultured and incubated with OMV for 48 hours in vitro, the secretion level of various inflammatory cytokines in Raw264.7 cells was detected by enzyme-linked immunosorbent assay (ELISA). The results showed that, compared with normal culture samples (normal-OMV), there was no significant difference in the morphology and size of OMV derived from P. aeruginosa under pressure culture with a certain concentration of MEM (MEM-OMV). The particle concentration of MEM-OMV was significantly higher than the particle concentration of Normal-OMV (20 ± 0.53×108/mL vs 1.50 ± 0.76×108/mL). The results of proteomics analysis showed that the protein expression of the two kind of OMVs was also heterogeneous, MEM-OMV containing more MexA and MexB proteins related to efflux pumps. The results of cell experiments showed that MEM-OMV could stimulate Raw264.7 cells to produce significantly higher concentrations of IL-6, TNF-α, IL-8 and IFN-γ than Normal-OMV. In conclusion, the addition of meropenem can change the production rate of OMV secreted by P. aeruginosa, the inclusion of protein and the immune stimulation effect on macrophages.

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Comparison in the characterization of outer membrane vesicles from Pseudomonas aeruginosa in normal culture and under meropenem pressure[J]. Journal of Microbes and Infections. 2024, 19(3): 129-139

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[1]CRONE S, VIVES-FLóREZ M, KVICH L, SAUNDERS A M, MALONE M, NICOLAISEN M H, MARTíNEZ-GARCíA E, ROJAS-ACOSTA C, CATALINA GOMEZ-PUERTO M, CALUM H, WHITELEY M, KOLTER R, BJARNSHOLT T.The environmental occurrence of Pseudomonas aeruginosa[J].Apmis, 2020, 128(3):220-231
[2]WANG K, CHEN Y Q, SALIDO M M, KOHLI G S, KONG J L, LIANG H J, YAO Z T, XIE Y T, WU H Y, CAI S Q, DRAUTZ-MOSES D I, DARLING A E, SCHUSTER S C, YANG L, DING Y.The rapid in vivo evolution of Pseudomonas aeruginosa in ventilator-associated pneumonia patients leads to attenuated virulence[J].Open Biol, 2017, 7(9):170029-170029
[3]ROSSI E, LA ROSA R, BARTELL J A, MARVIG R L, HAAGENSEN J A J, SOMMER L M, MOLIN S, JOHANSEN H K.Pseudomonas aeruginosa adaptation and evolution in patients with cystic fibrosis[J].Nat Rev Microbiol, 2021, 19(5):331-342
[4]JURADO-MARTíN I, SAINZ-MEJíAS M, MCCLEAN S.Pseudomonas aeruginosa: An Audacious Pathogen with an Adaptable Arsenal of Virulence Factors[J].International Journal of Molecular Sciences, 2021, 22(6):3128-3128
[5]JUODEIKIS R, CARDING SIMON R.Outer Membrane Vesicles: Biogenesis,Functions,and Issues[J].Microbiology and Molecular Biology Reviews, 2022, 86(4):e00032-00022
[6]BITTO N J, BAKER P J, DOWLING J K, WRAY-MCCANN G, DE PAOLI A, TRAN L S, LEUNG P L, STACEY K J, MANSELL A, MASTERS S L, FERRERO R L.Membrane vesicles from Pseudomonas aeruginosa activate the noncanonical inflammasome through caspase-5 in human monocytes[J].Immunol Cell Biol, 2018, 96(10):1120-1130
[7]ORENCH-RIVERA N, KUEHN M J.Environmentally controlled bacterial vesicle-mediated export[J].Cell Microbiol, 2016, 18(11):1525-1536
[8]METRUCCIO M M, EVANS D J, GABRIEL M M, KADURUGAMUWA J L, FLEISZIG S M.Pseudomonas aeruginosa Outer Membrane Vesicles Triggered by Human Mucosal Fluid and Lysozyme Can Prime Host Tissue Surfaces for Bacterial Adhesion[J].Front Microbiol, 2016, 7(1):871-871
[9]BAGGETT N S, BRONSON A S, CABEEN M T.SOS-Independent Pyocin Production in Paeruginosa Is Induced by XerC Recombinase Deficiency[J].mBio, 2021, 12(6):e0289321-e0289321
[10]MACDONALD I A, KUEHN M J.Stress-induced outer membrane vesicle production by Pseudomonas aeruginosa[J].J Bacteriol, 2013, 195(13):2971-2981
[11](CLSI) C A L S I.Performance standards for antimicrobial susceptibility testing[J].M100-Ed32 CLSI, 2022, (ISBN (electronic) )::, 978-971- 68440-68135-68442.-
[12]中华医学会呼吸病学分会感染学组.中国铜绿假单胞菌下呼吸道感染诊治专家共识（2022年版）[J].中华结核和呼吸杂志, 2022, 08(1):739-752
[13]LI Q, LI J, HE T, JI X, WEI R, YU M, WANG R.Sub-MIC Antibiotics Modulate Productions of Outer Membrane Vesicles in Tigecycline-Resistant Escherichia coli[J].Antibiotics (Basel), 2024, 13(3):276-276
[14]SARTORIO M G, PARDUE E J, FELDMAN M F, HAURAT M F.Bacterial Outer Membrane Vesicles: From Discovery to Applications[J].Annu Rev Microbiol, 2021, 75(1):609-630
[15]MAREDIA R, DEVINENI N, LENTZ P, DALLO S F, YU J, GUENTZEL N, CHAMBERS J, ARULANANDAM B, HASKINS W E, WEITAO T.Vesiculation from < i> Pseudomonas aeruginosa< /i> under SOS[J].The Scientific World Journal, 2012, :402919-
[16]PODLESEK Z, ?GUR BERTOK D.The DNA Damage Inducible SOS Response Is a Key Player in the Generation of Bacterial Persister Cells and Population Wide Tolerance[J].Front Microbiol, 2020, 11(1):1785-1785
[17]PAN Y P, XU Y H, WANG Z X, FANG Y P, SHEN J L.Overexpression of MexAB-OprM efflux pump in carbapenem-resistant Pseudomonas aeruginosa[J].Arch Microbiol, 2016, 198(6):565-571
[18]ARORA S K, RITCHINGS B W, ALMIRA E C, LORY S, RAMPHAL R.The Pseudomonas aeruginosa flagellar cap protein,FliD,is responsible for mucin adhesion[J].Infect Immun, 1998, 66(3):1000-1007
[19]WANG X, EAGEN W J, LEE J C.Orchestration of human macrophage NLRP3 inflammasome activation by Staphylococcus aureus extracellular vesicles[J].Proc Natl Acad Sci U S A, 2020, 117(6):3174-3184
[20]ZAVAN L, FANG H, JOHNSTON E L, WHITCHURCH C, GREENING D W, HILL A F, KAPARAKIS-LIASKOS M.The mechanism of Pseudomonas aeruginosa outer membrane vesicle biogenesis determines their protein composition[J].Proteomics, 2023, 23(10):e2200464-e2200464
[21]PARK K S, LEE J, JANG S C, KIM S R, JANG M H, L?TVALL J, KIM Y K, GHO Y S.Pulmonary inflammation induced by bacteria-free outer membrane vesicles from Pseudomonas aeruginosa[J].Am J Respir Cell Mol Biol, 2013, 49(4):637-645
[22]DEO P, CHOW S H, HAN M L, SPEIR M, HUANG C, SCHITTENHELM R B, DHITAL S, EMERY J, LI J, KILE B T, VINCE J E, LAWLOR K E, NADERER T.Mitochondrial dysfunction caused by outer membrane vesicles from Gram-negative bacteria activates intrinsic apoptosis and inflammation[J].Nat Microbiol, 2020, 5(11):1418-1427
[23]MCELVANEY O J, MCELVANEY N G.Targeting IL-8 in Cystic Fibrosis: Enough but Not Too Much[J].Am J Respir Cell Mol Biol, 2018, 59(4):401-402
[24]PRIEM B, VAN LEENT M M T, TEUNISSEN A J P, SOFIAS A M, MOURITS V P, WILLEMSEN L, KLEIN E D, OOSTERWIJK R S, MEERWALDT A E, MUNITZ J, PRéVOT G, VERA VERSCHUUR A, NAUTA S A, VAN LEEUWEN E M, FISHER E L, DE JONG K A M, ZHAO Y, TONER Y C, SOULTANIDIS G, CALCAGNO C, BOMANS P H H, FRIEDRICH H, SOMMERDIJK N, REINER T, DUIVENVOORDEN R, ZUPAN?I? E, DI MARTINO J S, KLUZA E, RASHIDIAN M, PLOEGH H L, DIJKHUIZEN R M, HAK S, PéREZ-MEDINA C, BRAVO-CORDERO J J, DE WINTHER M P J, JOOSTEN L A B, VAN ELSAS A, FAYAD Z A, RIALDI A, TORRE D, GUCCIONE E, OCHANDO J, NETEA M G, GRIFFIOEN A W, MULDER W J M.Trained Immunity-Promoting Nanobiologic Therapy Suppresses Tumor Growth and Potentiates Checkpoint Inhibition[J].Cell, 2020, 183(3):786-801
[25]LIU G, MA N, CHENG K, FENG Q, MA X, YUE Y, LI Y, ZHANG T, GAO X, LIANG J, ZHANG L, WANG X, REN Z, FU Y-X, ZHAO X, NIE G.Bacteria-derived nanovesicles enhance tumour vaccination by trained immunity[J].Nature Nanotechnology, 2023, 19(3):387-398