Staphylococcus aureus Phenol-Soluble Modulins Impair Interleukin Expression in Bovine Mammary Epithelial Cells

Deplanche, Martine; Алексеева, Л. И.; Semenovskaya, Ksenia; Fu, Chih-Lung; Dessauge, Frédéric; Finot, Laurence; Petzl, Wolfram; Zerbe, Holm; Loir, Yves Le; Rainard, Pascal; Smith, David George Emslie; Germon, Pierre; Otto, Michael; Berkova, Nadejda

doi:10.1128/iai.01330-15

Cited by 19 publications

(16 citation statements)

References 48 publications

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“…The MOI was based on pilot experiments that demonstrated that at this multiplicity, PS cells have the strongest IL-8 response to S. aureus in the absence of cytotoxic effects, as assessed by trypan blue staining and microscopy examination. The same maximum MOI for stimulating PS cells with S. aureus was also found by Deplanche et al [28]. After three hours, bacteria were removed by washing cells two times with PBS, followed by incubation in 0.5 mL SM containing 100 µg/mL streptomycin and 100 I.U./mL penicillin to prevent growth of bacteria.…”

Section: Ps Cell Responses To S Aureussupporting

confidence: 67%

Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent

et al. 2019

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Bovine mastitis is a costly disease to the dairy industry and intramammary infections (IMI) with Staphylococcus aureus are a major cause of mastitis. Staphylococcus aureus strains responsible for mastitis in cattle predominantly belong to ruminant-associated clonal complexes (CCs). Recognition of pathogens by bovine mammary epithelial cells (bMEC) plays a key role in activation of immune responsiveness during IMI. However, it is still largely unknown to what extent the bMEC response differs according to S. aureus CC. The aim of this study was to determine whether ruminant-associated S. aureus CCs differentially activate bMEC. For this purpose, the immortalized bMEC line PS was stimulated with S. aureus mastitis isolates belonging to four different clonal complexes (CCs; CC133, CC479, CC151 and CC425) and interleukin 8 (IL-8) release was measured as indicator of activation. To validate our bMEC model, we first stimulated PS cells with genetically modified S. aureus strains lacking (protein A, wall teichoic acid (WTA) synthesis) or expressing (capsular polysaccharide (CP) type 5 or type 8) factors expected to affect S. aureus recognition by bMEC. The absence of functional WTA synthesis increased IL-8 release by bMEC in response to bacterial stimulation compared to wildtype. In addition, bMEC released more IL-8 after stimulation with S. aureus expressing CP type 5 compared to CP type 8 or a strain lacking CP expression. Among the S. aureus lineages, isolates belonging to CC133 induced a significantly stronger IL-8 release from bMEC than isolates from the other CCs, and the IL-8 response to CC479 was higher compared to CC151 and CC425. Transcription levels of IL-8, tumor necrosis factor alpha (TNFα), serum amyloid A3 (SAA3), Toll-like receptor (TLR)-2 and nuclear factor κB (NF-κB) in bMEC after bacterial stimulation tended to follow a similar pattern as IL-8 release, but there were no significant differences between the CCs. This study demonstrates a differential activation of bMEC by ruminant-associated CCs of S. aureus, which may have implications for the severity of mastitis during IMI by S. aureus belonging to these lineages.

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Section: Ps Cell Responses To S Aureussupporting

confidence: 67%

Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent

et al. 2019

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“…For example, in conjunctiva epithelial cells, a significant increase in IL-8 mRNA was detected at 3 h post infection and a marked reduction after 4 h, an effect attributed to phenol-soluble modulins from S. aureus ( 45 ). Also, bovine mammary epithelial cells infected with S. aureus expressed IL-8 at initial stages (8 h), but at longer infection times (24–48 h), no increase was detected ( 46 ). In neutrophils, S. aureus strain USA300 inhibited NF-κB-dependent IL-8 expression and promoted cell death ( 6 ).…”

Section: Discussionmentioning

confidence: 99%

Glycogen Synthase Kinase 3α Is the Main Isoform That Regulates the Transcription Factors Nuclear Factor-Kappa B and cAMP Response Element Binding in Bovine Endothelial Cells Infected with Staphylococcus aureus

et al. 2018

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Glycogen synthase kinase 3 (GSK3) is a constitutive enzyme implicated in the regulation of cytokine expression and the inflammatory response during bacterial infections. Mammals have two GSK3 isoforms named GSK3α and GSK3β that plays different but often overlapping functions. Although the role of GSK3β in cytokine regulation during the inflammatory response caused by bacteria is well described, GSK3α has not been found to participate in this process. Therefore, we tested if GSK3α may act as a regulatory isoform in the cytokine expression by bovine endothelial cells infected with Staphylococcus aureus because this bacterium is one of the major pathogens that cause tissue damage associated with inflammatory dysfunction. Interestingly, although both isoforms were phosphorylated–inactivated, we consistently observed a higher phosphorylation of GSK3α at Ser21 than that of GSK3β at Ser9 after bacterial challenge. During a temporal course of infection, we characterized a molecular switch from pro-inflammatory cytokine expression (IL-8), promoted by nuclear factor-kappa B (NF-κB), at an early stage (2 h) to an anti-inflammatory cytokine expression (IL-10), promoted by cAMP response element binding (CREB), at a later stage (6 h). We observed an indirect effect of GSK3α activity on NF-κB activation that resulted in a low phosphorylation of CREB at Ser133, a decreased interaction between CREB and the co-activator CREB-binding protein (CBP), and a lower expression level of IL-10. Gene silencing of GSK3α and GSK3β with siRNA indicated that GSK3α knockout promoted the interaction between CREB and CBP that, in turn, increased the expression of IL-10, reduced the interaction of NF-κB with CBP, and reduced the expression of IL-8. These results indicate that GSK3α functions as the primary isoform that regulates the expression of IL-10 in endothelial cells infected with S. aureus.

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“…S. aureus produces phenol-soluble modulins (PSMs) as virulence factors, which have potent cytolytic activity against many cell types, including neutrophils and monocytes. Furthermore, PSMs decrease the expression of several defensin genes in epithelial cells and inhibit the production of IL-6, IL-8 and IL-32 in bovine MECs, resulting in attenuation of antibacterial function [ 3 ]. These components of S. uberis and S. aureus may prevent the bacteria from being attacked by immune cells and antimicrobial components.…”

Section: Discussionmentioning

confidence: 99%

Cellular and soluble components decrease the viable pathogen counts in milk from dairy cows with subclinical mastitis

Koshiishi¹,

Watanabe²,

Miyake³

et al. 2017

The Journal of Veterinary Medical Science

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The present study was undertaken to clarify the factors that reduce the viable pathogen count in milk collected from the udders of subclinical mastitic cows during preservation. Milk was centrifuged to divide somatic cells (cellular components, precipitates) and antimicrobial peptides (soluble components, supernatants without fat layer); each fraction was cultured with bacteria, and the number of viable bacteria was assessed prior to and after culture. In 28.8% of milk samples, we noted no viable bacteria immediately after collection; this value increased significantly after a 5-hr incubation of milk with cellular components but not with soluble components (48.1 and 28.8%, respectively). After culture with cellular components, the numbers of bacteria (excluding Staphylococcus aureus and Streptococcus uberis) and yeast decreased dramatically, although the differences were not statistically significant. After cultivation with soluble components, only yeasts showed a tendency toward decreased mean viability, whereas the mean bacterial counts of S. uberis and T. pyogenes tended to increase after 5-hr preservation with soluble components. These results suggest that most pathogens in high somatic cell count (SCC) milk decreased during preservation at 15 to 25°C, due to both the cellular components and antimicrobial components in the milk. Particularly, the cellular components more potently reduced bacterial counts during preservation.

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Staphylococcus aureus Phenol-Soluble Modulins Impair Interleukin Expression in Bovine Mammary Epithelial Cells

Cited by 19 publications

References 48 publications

Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent

Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent

Glycogen Synthase Kinase 3α Is the Main Isoform That Regulates the Transcription Factors Nuclear Factor-Kappa B and cAMP Response Element Binding in Bovine Endothelial Cells Infected with Staphylococcus aureus

Cellular and soluble components decrease the viable pathogen counts in milk from dairy cows with subclinical mastitis

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