Molecular basis of surface anchored protein A deficiency in the Staphylococcus aureus strain Wood 46

Balachandran, Manasi; Giannone, Richard J.; Bemis, David A.; Kania, Stephen A.

doi:10.1371/journal.pone.0183913

Cited by 7 publications

(5 citation statements)

References 60 publications

(71 reference statements)

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“…Because we observed background binding of control IgG1 to Wood46 biofilm compared to planktonic Wood46 ( Figure 2—figure supplement 1 ), we wondered whether this could be explained by secreted SpA being incorporated in the biofilm matrix as Wood46 is unable to link secreted SpA to the surface due to a sortase defect ( Balachandran et al, 2017a ). To test this hypothesis, we performed a binding assay on planktonic versus biofilm Wood46 using nonspecific IgG1, nonspecific IgG3 (which is unable to bind to SpA via the Fc-domain Jendeberg et al, 1997 ), and anti-SpA-IgG3 (binding SpA via the Fab-domain but not the Fc-domain).…”

Section: Resultsmentioning

confidence: 99%

“…S. aureus strains Wood46 (ATCC 10832) ( Amend et al, 1984 ; Balachandran et al, 2017b ; Balachandran et al, 2017a ), USA300 LAC (AH1263) ( Boles et al, 2010 ), and USA300 LAC ∆spa, sbi ::Tn (AH4116) were used in this study. Strain USA300 LAC ∆spa, sbi ::Tn (AH4116) was constructed by transducing sbi ::Tn from Nebraska Transposon Library ( Fey et al, 2013 ) into USA300 LAC ∆spa (AH3052) ( Ibberson et al, 2014 ) with phage 11.…”

Section: Methodsmentioning

confidence: 99%

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Human monoclonal antibodies against Staphylococcus aureus surface antigens recognize in vitro and in vivo biofilm

Vor

Dijk

Kessel

et al. 2022

eLife

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Implant-associated Staphylococcus aureus infections are difficult to treat because of biofilm formation. Bacteria in a biofilm are often insensitive to antibiotics and host immunity. Monoclonal antibodies (mAbs) could provide an alternative approach to improve the diagnosis and potential treatment of biofilm-related infections. Here, we show that mAbs targeting common surface components of S. aureus can recognize clinically relevant biofilm types. The mAbs were also shown to bind a collection of clinical isolates derived from different biofilm-associated infections (endocarditis, prosthetic joint, catheter). We identify two groups of antibodies: one group that uniquely binds S. aureus in biofilm state and one that recognizes S. aureus in both biofilm and planktonic state. Furthermore, we show that a mAb recognizing wall teichoic acid (clone 4497) specifically localizes to a subcutaneously implanted pre-colonized catheter in mice. In conclusion, we demonstrate the capacity of several human mAbs to detect S. aureus biofilms in vitro and in vivo.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Human monoclonal antibodies against Staphylococcus aureus surface antigens recognize in vitro and in vivo biofilm

Vor

Dijk

Kessel

et al. 2022

eLife

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“…Inhibition of SrtA activity may therefore have a different, strain-dependent effect of changing the surface expression of the modified bacterial proteins [36,37]. For example a portion of SpA is released into the supernatant with an intact sorting signal, and release of SpA was reduced when the native sorting signal of SpA was replaced with the corresponding region of another sortase-anchored protein, namely, SdrE [38].…”

Section: Resultsmentioning

confidence: 99%

The Pros and Cons of Cystic Fibrosis (CF) Patient Use of Herbal Supplements Containing Pulmonaria officinalis L. Extract: the Evidence from an In Vitro Study on Staphylococcus aureus CF Clinical Isolates

Sadowska

Wójcik

Krzyżanowska-Kowalczyk

et al. 2019

Molecules

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The justification for the use of herbal supplements with Pulmonaria officinalis L. extract (POE) in the case of staphylococcal lung colonization/infections characteristic for cystic fibrosis (CF), was examined in vitro. The impact of POE phenolic-rich fraction on the virulence attributes of CF-associated Staphylococcus aureus (S. aureus) clinical strains has been assessed, including pathogen adhesion, biofilm formation on native and protein-conditioned surfaces (mucin, elastin), mature biofilm eradication, staphylococcal protein A expression, α-toxin release, and S. a. adhesion to A549 cells. Cytotoxicity of the extract to lung epithelial cells was also investigated. It was found that POE has bacteriostatic effects at MIC 1–2 mg/mL, recognized as of limited efficacy, but at MIC/subMICs it targeted virulence not viability. It usually decreased S. aureus adhesion and less frequently inhibited biofilm formation on native and protein-conditioned surfaces. Observed effect seems to be related to significant reduction by POE of sortase A activity. However, in some cases POE favored the creation of biofilm by staphylococci and S. aureus adhesion to the lung epithelium was not limited. On the other side POE caused significant decrease of S. a. α-toxin synthesis and slightly weakened the expression of SpA. When used at supraMICs POE eradicated mature biofilm, but in some cases with unsatisfying outcomes. Promisingly, POE has been recognized as a safe product, with no cytotoxicity up to 4 mg/mL. These results reflect the positive, negative or neutral anti-staphylococcal properties of POE. It seems that POE may be beneficial as a prophylactic, but not as a therapeutic or supportive agent in the area of CF—integrative medicine. However, introduction the official recommendations needs further in vivo studies.

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“…Большое количество штаммов стафилококка продуцирует протеин А, который получают путем лизиса бактериальной клетки или из супернатанта культуральной жидкости. Штамм S. aureus Cowan I известен как хороший продуцент белка А, а штамм Wood 46 характеризуется отсутствием гена spa, кодирующего экспрессию белка А [24,25]. Соответственно, водорастворимые АГ S. aureus Wood 46 не взаимодействовали с МкАТ (rPly) -ПХ.…”

Section: Discussionunclassified

Development of the test system based on enzyme immunoassay for the detection of recombinant <i>Streptococcus</i> pneumoniae pneumolysin

Kurbatova,

Yakovleva,

Gavrilova

et al. 2023

Rossijskij bioterapevtičeskij žurnal

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Backgraund. Pneumolysin (Ply) is a hemolytic toxin of Streptococcus pneumoniae (S. pneumoniae) expressed by all strains of pneumococci. The use of sandwich enzyme-linked immunosorbent assay (ELISA) can be a simple, fast and effective way of its qualitative and quantitative determination in biological fluids.Aim. To develop and evaluate the specificity of sandwich ELISA test system for qualitative and quantitative determination of recombinant Ply (rPly) of S. pneumoniae.Materials and methods. Immobilized on the solid phase rabbit’s polyclonal antibodies (pAbs) to rPly were used as recognition antibodies in sandwich ELISA. The studied antigens were added to the pAbs (rPly). The reaction was manifested by using detecting mouse monoclonal IgG1 (rPly) – antibodies conjugated with horseradish root peroxidase. The specificity of the test system was evaluated when using recombinant α-hemolysin (rα-Hly) and water-soluble S. aureus antigens as reference preparations.Results. Using sandwich ELISA, rPly was detected at a concentration of 0.15 µ / ml. The test system was characterized by specificity, which was confirmed by the absence of reaction with recombinant rα-Hly of Staphylococcus aureus (S. aureus). Reference preparations of water-soluble surface antigens of S. aureus strains No 209, 1986,1991 and Cowan I gave a false positive reaction due to the presence of protein A (SpA) in their composition, a thermostable surface protein expressed by many strains of staphylococci capable of binding immunoglobulins via Fc-fragment or Fab fragments of the V3H domain of the B cells receptor. A negative reaction was obtained with antigens from the S. aureus wood 46 strain, which does not have the spa gene encoding SpA expression. The presence of protein A in preparations of water-soluble S. aureus antigens was confirmed in the ELISA inhibition assay.Conclusion. Sandwich ELISA has been developed for qualitative and quantitative determination of S. pneumoniae Ply. The conducted studies have confirmed the specificity of the test system.

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Molecular basis of surface anchored protein A deficiency in the Staphylococcus aureus strain Wood 46

Cited by 7 publications

References 60 publications

Human monoclonal antibodies against Staphylococcus aureus surface antigens recognize in vitro and in vivo biofilm

Human monoclonal antibodies against Staphylococcus aureus surface antigens recognize in vitro and in vivo biofilm

The Pros and Cons of Cystic Fibrosis (CF) Patient Use of Herbal Supplements Containing Pulmonaria officinalis L. Extract: the Evidence from an In Vitro Study on Staphylococcus aureus CF Clinical Isolates

Development of the test system based on enzyme immunoassay for the detection of recombinant <i>Streptococcus</i> pneumoniae pneumolysin

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