title: Differential Responses of Pattern Recognition Receptors to Outer Membrane Vesicles of Three Periodontal Pathogens.
abstract: Highly purified outer membrane vesicles (OMVs) of the periodontal pathogens, Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia were produced using tangential flow ultrafiltration, ultracentrifugation and Optiprep density gradient separation. Cryo-TEM and light scattering showed OMVs to be single lipid-bilayers with modal diameters of 75 to 158 nm. Enumeration of OMVs by nanoparticle flow-cytometry at the same stage of late exponential culture indicated that P. gingivalis was the most prolific OMV producer. P. gingivalis OMVs induced strong TLR2 and TLR4-specific responses and moderate responses in TLR7, TLR8, TLR9, NOD1 and NOD2 expressing-HEK-Blue cells. Responses to T. forsythia OMVs were less than those of P. gingivalis and T. denticola OMVs induced only weak responses. Compositional analyses of OMVs from the three pathogens demonstrated differences in protein, fatty acids, lipopolysaccharide, peptidoglycan fragments and nucleic acids. Periodontal pathogen OMVs induced differential pattern recognition receptor responses that have implications for their role in chronic periodontitis.
authors.affiliation: Oral Health Cooperative Research Centre, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Melbourne, Victoria, Australia.
authors.forename: Jessica D
authors.initials: JD
authors.lastname: Cecil
authors.affiliation: Oral Health Cooperative Research Centre, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Melbourne, Victoria, Australia.
authors.forename: Neil M
authors.initials: NM
authors.lastname: O'Brien-Simpson
authors.affiliation: Oral Health Cooperative Research Centre, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Melbourne, Victoria, Australia.
authors.forename: Jason C
authors.initials: JC
authors.lastname: Lenzo
authors.affiliation: Oral Health Cooperative Research Centre, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Melbourne, Victoria, Australia.
authors.forename: James A
authors.initials: JA
authors.lastname: Holden
authors.affiliation: Oral Health Cooperative Research Centre, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Melbourne, Victoria, Australia.
authors.forename: Yu-Yen
authors.initials: YY
authors.lastname: Chen
authors.affiliation: Oral Health Cooperative Research Centre, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Melbourne, Victoria, Australia.
authors.forename: William
authors.initials: W
authors.lastname: Singleton
authors.affiliation: Department of Chemical and Biomolecular Engineering, The University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Katelyn T
authors.initials: KT
authors.lastname: Gause
authors.affiliation: Department of Chemical and Biomolecular Engineering, The University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Yan
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Department of Chemical and Biomolecular Engineering, The University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Frank
authors.initials: F
authors.lastname: Caruso
authors.affiliation: Oral Health Cooperative Research Centre, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Melbourne, Victoria, Australia.
authors.forename: Eric C
authors.initials: EC
authors.lastname: Reynolds
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D001439:Bacteroides / Q000737:chemistry / Q000276:immunology*; D002460:Cell Line; D055113:Chronic Periodontitis / Q000276:immunology* / Q000382:microbiology; D006801:Humans; D008070:Lipopolysaccharides / Q000032:analysis; D053474:Nod1 Signaling Adaptor Protein / Q000276:immunology; D053473:Nod2 Signaling Adaptor Protein / Q000276:immunology; D010457:Peptidoglycan / Q000032:analysis; D016966:Porphyromonas gingivalis / Q000737:chemistry / Q000276:immunology*; D051192:Receptors, Pattern Recognition / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D051195:Toll-Like Receptor 2 / Q000276:immunology; D051197:Toll-Like Receptor 4 / Q000276:immunology; D048373:Treponema denticola / Q000737:chemistry / Q000276:immunology*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D008070:Lipopolysaccharides; C119067:NOD1 protein, human; C429018:NOD2 protein, human; D053474:Nod1 Signaling Adaptor Protein; D053473:Nod2 Signaling Adaptor Protein; D010457:Peptidoglycan; D051192:Receptors, Pattern Recognition; C495342:TLR2 protein, human; C495345:TLR4 protein, human; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4