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Extracellular vesicles from gram-positive bacteria Staphylococcus aureus and Staphylococcus epidermidis induce a strong inflammatory response and cell death in vitro

Poster (konferens)
Författare Forugh Vazirisani
Margarita Trobos
Magdalena Zaborowska
Xiaoqin Wang
Omar Omar
Karin Ekström
Peter Thomsen
Publicerad i ISEV 2016, Rotherdam, Netherlands
Publiceringsår 2016
Publicerad vid Institutionen för kliniska vetenskaper, Avdelningen för biomaterialvetenskap
Språk en
Ämneskategorier Mikrobiologi

Sammanfattning

The majority of biomaterial-associated infections (BAI) are caused by S.aureus and S.epidermidis. Bacterial extracellular vesicles (EVs) are involved in the delivery of toxins and bacterial components to host cells. It is therefore of relevance to investigate if the EVs from these bacterial species have a role in BAI. The aim of the present study was: i) to investigate whether clinical strains from S. aureus and S. epidermidis, isolated from osteomyelitis associated with bone-anchored amputation prosthesis, release EVs, ii) to study the biological effects of these EVs on monocytes and iii) to evaluate the effect of these EVs on the secretion of pro-inflammatory substances by monocytes. Material & Methods EVs were isolated from S. aureus 64516 and S. epidermidis 64518 cultures. The collected EVs were characterized by Western blot. Human primary monocytes were stimulated by different concentrations of EVs (0, 5, 10 and 50 µg/ml). After 24 h, cell death was evaluated using propidium iodide in a nucleocounter and further, levels of IL-8, IL-6, TNF-α, MCP-1 and MMP-9 were determined by ELISA. Results The two staphylococcal strains secreted EVs in vitro. Both types of EVs contained δ-toxin whereas protein A and SCP-A were only detected in S. aureus EVs. The monocyte viability was reduced in a dose-dependent manner after incubation with EVs. More than 50 % of monocytes died with 5 µg S. aureus EVs. The S. epidermidis EVs had relatively less toxic effects. Stimulation of monocytes with S. aureus and S. epidermidis EVs significantly increased the release of IL-8, IL-6, TNF-α, MCP-1 and MMP-9. Conclusion The results from this study show that clinical pathogens causing BAI have the ability to secrete EVs in vitro. The EVs promote pro-inflammatory cytokine release from human monocyte and subsequent cell death. It is suggested that EVs contribute to the inflammation and injury associated with biomaterials-associated infection.

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