Cytokine induction of peripheral blood mononuclear cells by biofilms and biofilm supernatants of Granulicatella and Abiotrophia spp.

Bhardwaj, Radhika G.; Al-Khabbaz, Areej K.; Karched, Maribasappa

doi:10.1016/j.micpath.2017.11.037

Cited by 15 publications

(10 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although cytokine induction of PBMCs from total protein preparation from G. adiacens was similar, MCP-1 and G-CSF were absent. This was also observed in our previous study where biofilms failed to stimulate these cytokines but the biofilm supernatants did (Bhardwaj et al, 2018). While secreted components of different bacterial species have been previously shown to elicit inflammatory response from host cells (Oscarsson et al, 2008; Dapunt et al, 2016), a protein of Brucella suis was able to inhibit TNF-α production from macrophages when it was released extracellularly (Caron et al, 1996).…”

Section: Discussionsupporting

confidence: 87%

“…Ethical approval for blood collection from a healthy human volunteer was obtained from Health Science Center Ethical Committee, Kuwait University. Peripheral blood mononuclear cells (PBMCs) were isolated from the blood of a systemically healthy human volunteer as described earlier (Fuss et al, 2009; Bhardwaj et al, 2018). Blood was collected by venipuncture into tubes containing heparin vacutainer (4 ml /tube).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Proteomic Analysis and Virulence Assessment of Granulicatella adiacens Secretome

Karched

Bhardwaj

Tiss

et al. 2019

Front. Cell. Infect. Microbiol.

Self Cite

View full text Add to dashboard Cite

Despite reports on the occurrence of Granulicatella adiacens in infective endocarditis, few mechanistic studies on its virulence characteristics or pathogenicity are available. Proteins secreted by this species may act as determinants of host-microbe interaction and play a role in virulence. Our aim in this study was to investigate and functionally characterize the secretome of G. adiacens . Proteins in the secretome preparation were digested by trypsin and applied to nanoLC-ESI-MS/MS. By using a combined mass spectrometry and bioinformatics approach, we identified 101 proteins. Bioinformatics tools predicting subcellular localization revealed that 18 of the secreted proteins possessed signal sequence. More than 20% of the secretome proteins were putative virulence proteins including serine protease, superoxide dismutase, aminopeptidase, molecular chaperone DnaK, and thioredoxin. Ribosomal proteins, molecular chaperones, and glycolytic enzymes, together known as “moonlighting proteins,” comprised fifth of the secretome proteins. By Gene Ontology analysis, more than 60 proteins of the secretome were grouped in biological processes or molecular functions. KEGG pathway analysis disclosed that the secretome consisted of enzymes involved in biosynthesis of antibiotics. Cytokine profiling revealed that secreted proteins stimulated key cytokines, such as IL-1β, MCP-1, TNF-α, and RANTES from human PBMCs. In summary, the results from the current investigation of the G. adiacens secretome provide a basis for understanding possible pathogenic mechanisms of G. adiacens .

show abstract

Section: Discussionsupporting

confidence: 87%

Section: Methodsmentioning

confidence: 99%

Proteomic Analysis and Virulence Assessment of Granulicatella adiacens Secretome

Karched

Bhardwaj

Tiss

et al. 2019

Front. Cell. Infect. Microbiol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…They are all gram‐positive, nonmotile, nonspore forming, and can be considered commensal species of the gut microbiota. Importantly, they are also aero‐tolerant or facultatively anaerobic (Bhardwaj, Al‐Khabbaz, & Karched, ; Davis, ; Murray, ; Patterson, ). These findings represent a marked difference between this observation of the C. apella gut microbiota and what is known regarding the human gut microbiota.…”

Section: Discussionmentioning

confidence: 99%

Metagenomic assessment of the Cebus apella gut microbiota

Firrman

Liu

Tanes

et al. 2019

American J Primatol

View full text Add to dashboard Cite

Cebus Apella (C. apella) is a species of Nonhuman Primate (NHP) used for biomedical research because it is phylogenetically similar and shares anatomical commonalities with humans. Here, the gut microbiota of three C. apella were examined in the different regions of the intestinal tract. Using metagenomics, the gut microbiota associated with the luminal content and mucus layer for each intestinal region was identified, and functionality was investigated by quantifying the levels of short chain fatty acids (SCFAs) produced. The results of this study show a high degree of similarity in the intestinal communities among C. apella subjects, with multiple shared characteristics. First, the communities in the lumen were more phylogenetically diverse and rich compared to the mucus layer communities throughout the entire intestinal tract. The small intestine communities in the lumen displayed a higher Shannon diversity index compared to the colon communities. Second, all the communities were dominated by aero-tolerant taxa such as Streptococcus, Enterococcus, Abiotrophia, and Lactobacillus, although there was preferential colonization of specific taxa observed. Finally, the primary SCFA produced throughout the intestinal tract was acetic acid, with some propionic acid and butyric acid detected in the colon regions. The small intestine microbiota produced significantly less SCFAs compared to the communities in the colon. Collectively, these data provide an in-depth report on the composition, distribution, and SCFA production of the gut microbiota along the intestinal tract of the C. apella NHP animal model. K E Y W O R D SCebus apella, gut microbiota, in vivo studies, intestinal tract, primate research, tufted capuchin

show abstract

“…PBMCs from the blood of a healthy human volunteer were isolated using Ficoll-Paque density gradient centrifugation method [35]. After obtaining written informed consent from the donor, blood was collected by venipuncture into vacutainer heparin tubes (3 ml per tube).…”

Section: Methodsmentioning

confidence: 99%

Proteomics of Extracellular Vesicles Produced byGranulicatellaSpecies that cause Infective Endocarditis

Ellepola

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

30When oral bacteria accidentally enter the bloodstream due to transient tissue 31 damage during dental procedures, they have the potential to attach to the endocardium 32 or an equivalent surface of an indwelling prosthesis and cause infection. Many bacterial 33 species produce extracellular vesicles (EVs) as part of normal physiology, but also use 34 it as a virulence strategy. In this study, it was hypothesized that Granulicatella species 35 produce EVs that possibly help them in virulence. Therefore, the objectives were to 36 isolate and characterize EVs produced by these species and to investigate their immune-37 stimulatory effects. The reference strains G. adiacens CCUG 27809 and G. elegans 38 CCUG 38949 were cultured on chocolate blood agar for 2 days. From subsequent broth 39 cultures, the EVs were isolated using differential centrifugation and filtration protocol 40 and then observed using scanning electron microscopy. Proteins in the vesicle 41 preparations were identified by nano LC-ESI-MS/MS. The EVs proteomes were 42 analyzed and characterized using different bioinformatics tools. The immune-43 stimulatory effect of the EVs was studied via ELISA quantification of IL-8, IL-1β and 44 CCL5, major proinflammatory cytokines, produced from stimulated human PBMCs. It 45 was revealed that both G. adiacens and G. elegans produced EVs, ranging in diameter 46 from 30 to 250 nm. Overall, G. adiacens EVs contained 160 proteins, and G. elegans 47 EVs contained 107 proteins. Both proteomes consist of several ribosomal proteins, 48DNA associated proteins, binding proteins, and metabolic enzymes. It was also shown 49 that these EVs carry putative virulence factors including moonlighting proteins. These 50 EVs were able to induce the production of IL-8, IL-1β and CCL5 from human PBMCs. 51The diversity in EVs content indicates that these vesicles could have possible roles in 3 52 bacterial survival, invasion, host immune modulation as well as infection. Further 53 functional characterization of the Granulicatella EVs may provide new insights into 54 virulence mechanisms of these important but less studied oral bacterial species. 55 56 57 Granulicatella species, formerly known as nutritionally variant streptococci 58 based on their characteristic dependence on pyridoxal or cysteine supplementation for 59 their growth in standard media [1], are catalase and oxidase negative, non-motile, non-60 spore-forming, facultatively anaerobic Gram-positive cocci [2, 3]. They are part of the 61 normal oral flora [4], but cause serious infections such as infective endocarditis. The 62 genus Granulicatella consists of 3 species: Granulicatella adiacens, Granulicatella 63 elegans and Granulicatella balaenopterae [3]. The species G. balaenopterae has not 64 been isolated from human samples, whereas both G. adiacens and G. elegans have been 65 reported from IE cases [5, 6]. In addition, these oral commensal cocci have been 66 associated with endodontic infections [7, 8], dental caries [9], and periodontitis [8, 10] 67 via DNA-based s...

show abstract

Cytokine induction of peripheral blood mononuclear cells by biofilms and biofilm supernatants of Granulicatella and Abiotrophia spp.

Cited by 15 publications

References 39 publications

Proteomic Analysis and Virulence Assessment of Granulicatella adiacens Secretome

Proteomic Analysis and Virulence Assessment of Granulicatella adiacens Secretome

Metagenomic assessment of the Cebus apella gut microbiota

Proteomics of Extracellular Vesicles Produced byGranulicatellaSpecies that cause Infective Endocarditis

Contact Info

Product

Resources

About