scholarly journals Inhibition of Biofilm Formation by Monoclonal Antibodies against Staphylococcus epidermidis RP62A Accumulation-Associated Protein

2005 ◽  
Vol 12 (1) ◽  
pp. 93-100 ◽  
Author(s):  
Daqian Sun ◽  
M. A. Accavitti ◽  
J. D. Bryers
Keyword(s):  
Cell Wall ◽  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Polyclonal Antibodies ◽  
Bacterial Colonization ◽  
Reversed Phase ◽  
Negative Control ◽  
Protein Accumulation ◽  
Dynamic Expression ◽  
Accumulation Associated Protein

ABSTRACT Staphylococcus epidermidis expresses a 140-kDa cell wall-bound protein accumulation-associated protein (AAP) to adhere to and accumulate as a biofilm on a surface. Potentially blocking AAP with a monoclonal antibody (MAb) could reduce or eliminate S. epidermidis bacterial colonization of biomedical devices. Here, we report on our efforts to (i) isolate AAP, (ii) generate MAbs against AAP, and (iii) determine the efficacy of MAbs to inhibit S. epidermidis biofilm formation. An M7 S. epidermidis mutant, reportedly deficient in AAP expression, was used as a negative control. Postinoculation murine sera, containing polyclonal antibodies against AAP, were able to reduce S. epidermidis biofilm formation by 54%. Select MAbs against AAP were able to reduce S. epidermidis by no more than 66%. Two MAb mixtures, 12C6/12A1 and 3C1/12A1, reduced S. epidermidis accumulation up to 79 and 87%, respectively, significantly more than individual MAbs. Contrary to a previous report, biofilm-deficient S. epidermidis mutant M7 expressed a 200-kDa protein on its cell wall that specifically bound AAP MAbs. Peptide characterization of this M7 protein by microcapillary reversed-phase high-pressure liquid chromatography-nanoelectrospray tandem mass spectrometry resulted in 53% homology with AAP. Ongoing studies will elucidate the dynamic expression of AAP and the M7 200-kDa protein in order to define their roles in biofilm formation.

scholarly journals A Single B-Repeat of Staphylococcus epidermidis Accumulation-Associated Protein Induces Protective Immune Responses in an Experimental Biomaterial-Associated Infection Mouse Model

2014 ◽  
Vol 21 (9) ◽  
pp. 1206-1214 ◽  
Author(s):  
Lin Yan ◽  
Lei Zhang ◽  
Hongyan Ma ◽  
David Chiu ◽  
James D. Bryers
Keyword(s):  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Porous Scaffold ◽  
The United States ◽  
Protein Vaccine ◽  
Weight Changes ◽  
Biofilm Inhibition ◽  
Content Type ◽  
Accumulation Associated Protein

ABSTRACTNosocomial infections are the fourth leading cause of morbidity and mortality in the United States, resulting in 2 million infections and ∼100,000 deaths each year. More than 60% of these infections are associated with some type of biomedical device.Staphylococcus epidermidisis a commensal bacterium of the human skin and is the most common nosocomial pathogen infecting implanted medical devices, especially those in the cardiovasculature.S. epidermidisantibiotic resistance and biofilm formation on inert surfaces make these infections hard to treat. Accumulation-associated protein (Aap), a cell wall-anchored protein ofS. epidermidis, is considered one of the most important proteins involved in the formation ofS. epidermidisbiofilm. A small recombinant protein vaccine comprising a single B-repeat domain (Brpt1.0) ofS. epidermidisRP62A Aap was developed, and the vaccine's efficacy was evaluatedin vitrowith a biofilm inhibition assay andin vivoin a murine model of biomaterial-associated infection. A high IgG antibody response againstS. epidermidisRP62A was detected in the sera of the mice after two subcutaneous immunizations with Brpt1.0 coadministered with Freund's adjuvant. Sera from Brpt1.0-immunized mice inhibitedin vitroS. epidermidisRP62A biofilm formation in a dose-dependent pattern. After receiving two immunizations, each mouse was surgically implanted with a porous scaffold disk containing 5 × 106CFU ofS. epidermidisRP62A. Weight changes, inflammatory markers, and histological assay results after challenge withS. epidermidisindicated that the mice immunized with Brpt1.0 exhibited significantly higher resistance toS. epidermidisRP62A implant infection than the control mice. Day 8 postchallenge, there was a significantly lower number of bacteria in scaffold sections and surrounding tissues and a lower residual inflammatory response to the infected scaffold disks for the Brpt1.0-immunized mice than for of the ovalbumin (Ova)-immunized mice.

scholarly journals Accumulation-Associated Protein Enhances Staphylococcus epidermidis Biofilm Formation under Dynamic Conditions and Is Required for Infection in a Rat Catheter Model

2014 ◽  
Vol 83 (1) ◽  
pp. 214-226 ◽  
Author(s):  
Carolyn R. Schaeffer ◽  
Keith M. Woods ◽  
G. Matt Longo ◽  
Megan R. Kiedrowski ◽  
Alexandra E. Paharik ◽  
...  
Keyword(s):  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Host Cells ◽  
Content Type ◽  
Microtiter Plates ◽  
Allelic Replacement ◽  
Abiotic Surfaces ◽  
Cell Accumulation ◽  
Accumulation Associated Protein

Biofilm formation is the primary virulence factor ofStaphylococcus epidermidis.S. epidermidisbiofilms preferentially form on abiotic surfaces and may contain multiple matrix components, including proteins such as accumulation-associated protein (Aap). Following proteolytic cleavage of the A domain, which has been shown to enhance binding to host cells, B domain homotypic interactions support cell accumulation and biofilm formation. To further define the contribution of Aap to biofilm formation and infection, we constructed anaapallelic replacement mutant and anicaADBC aapdouble mutant. When subjected to fluid shear, strains deficient in Aap production produced significantly less biofilm than Aap-positive strains. To examine thein vivorelevance of our findings, we modified our previously described rat jugular catheter model and validated the importance of immunosuppression and the presence of a foreign body to the establishment of infection. The use of our allelic replacement mutants in the model revealed a significant decrease in bacterial recovery from the catheter and the blood in the absence of Aap, regardless of the production of polysaccharide intercellular adhesin (PIA), a well-characterized, robust matrix molecule. Complementation of theaapmutant with full-length Aap (containing the A domain), but not the B domain alone, increased initial attachment to microtiter plates, as did intransexpression of the A domain in adhesion-deficientStaphylococcus carnosus. These results demonstrate Aap contributes toS. epidermidisinfection, which may in part be due to A domain-mediated attachment to abiotic surfaces.

Overview of Staphylococcus epidermidis cell wall-anchored proteins: potential targets to inhibit biofilm formation

2019 ◽  
Vol 47 (1) ◽  
pp. 771-784 ◽  
Author(s):  
Silvestre Ortega-Peña ◽  
Sergio Martínez-García ◽  
Sandra Rodríguez-Martínez ◽  
Mario E. Cancino-Diaz ◽  
Juan C. Cancino-Diaz
Keyword(s):  
Cell Wall ◽  
Staphylococcus Epidermidis ◽  
Biofilm Formation

scholarly journals Inhibition of Staphylococcus epidermidis Biofilm Formation by Rabbit Polyclonal Antibodies against the SesC Protein

2009 ◽  
Vol 77 (9) ◽  
pp. 3670-3678 ◽  
Author(s):  
Mohammad Shahrooei ◽  
Vishal Hira ◽  
Benoit Stijlemans ◽  
Rita Merckx ◽  
Peter W. M. Hermans ◽  
...  
Keyword(s):  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Polyclonal Antibodies ◽  
Late Phase ◽  
Surface Protein ◽  
Lpxtg Motif ◽  
Fibrinogen Binding ◽  
Gene And Protein Expression

ABSTRACT Several well-studied proteins with defined roles in Staphylococcus epidermidis biofilm formation are LPXTG motif-containing proteins. Here, we investigate the possible use of the LPXTG motif-containing protein SesC (S . epidermidis surface protein C; accession no. NP_765787) as a target for antibodies to prevent biofilm formation. In vitro and in a in vivo rat model of catheter infection, gene and protein expression analysis showed that SesC is expressed more strongly in biofilm-associated cells than in planktonic cells and is expressed particularly during the late phase of in vivo biofilm formation. Polyclonal rabbit antibodies raised against SesC reduced the fibrinogen-binding ability of S. epidermidis RP62A and Staphylococcus aureus RN4220 transformants expressing SesC, inhibited in vitro biofilm formation by S. epidermidis strains 10b and 1457, and significantly reduced the numbers of bacteria in a 1-day-old in vivo biofilm (P < 0.001, one-way analysis of variance). Our findings revealed that SesC is a promising target for prevention and treatment of S. epidermidis biofilms because it affects both the primary attachment and biofilm accumulation phases. The precise role of SesC in biofilm formation remains to be identified.

scholarly journals The Terminal A Domain of the Fibrillar Accumulation-Associated Protein (Aap) of Staphylococcus epidermidis Mediates Adhesion to Human Corneocytes

2009 ◽  
Vol 191 (22) ◽  
pp. 7007-7016 ◽  
Author(s):  
Robin L. Macintosh ◽  
Jane L. Brittan ◽  
Ritwika Bhattacharya ◽  
Howard F. Jenkinson ◽  
Jeremy Derrick ◽  
...  
Keyword(s):  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Domain Architecture ◽  
Opportunistic Pathogen ◽  
Additional Function ◽  
Artificial Joints ◽  
A Cell ◽  
Skin Colonization ◽  
Accumulation Associated Protein

ABSTRACT The opportunistic pathogen Staphylococcus epidermidis colonizes indwelling medical devices by biofilm formation but is primarily a skin resident. In many S. epidermidis strains biofilm formation is mediated by a cell wall-anchored protein, the accumulation-associated protein (Aap). Here, we investigate the role of Aap in skin adhesion. Aap is an LPXTG protein with a domain architecture including a terminal A domain and a B-repeat region. S. epidermidis NCTC 11047 expresses Aap as localized, lateral tufts of fibrils on one subpopulation of cells (Fib+), whereas a second subpopulation does not express these fibrils of Aap (Fib−). Flow cytometry showed that 72% of NCTC 11047 cells expressed Aap and that 28% of cells did not. Aap is involved in the adhesion of Fib+ cells to squamous epithelial cells from the hand (corneocytes), as the recombinant A-domain protein partially blocked binding to corneocytes. To confirm the role of the Aap A domain in corneocyte attachment, Aap was expressed on the surface of Lactococcus lactis MG1363 as sparsely distributed, peritrichous fibrils. The expression of Aap increased corneocyte adhesion 20-fold compared to L. lactis carrying Aap without an A domain. S. epidermidis isolates from catheters, artificial joints, skin, and the nose also used the A domain of Aap to adhere to corneocytes, emphasizing the role of Aap in skin adhesion. In addition, L. lactis expressing Aap with different numbers of B repeats revealed a positive correlation between the number of B repeats and adhesion to corneocytes, suggesting an additional function for the B region in enhancing A-domain-dependent attachment to skin. Therefore, in addition to its established role in biofilm formation, Aap can also promote adhesion to corneocytes and is likely to be an important adhesin in S. epidermidis skin colonization.

Quantitative Analysis of Biofilm Formed on Vascular Prostheses by Staphylococcus Epidermidis with Different ica and aap Genetic Status

2013 ◽  
Vol 36 (2) ◽  
pp. 105-112 ◽  
Author(s):  
Joanna Golus ◽  
Magdalena Stankevic ◽  
Rafal Sawicki ◽  
Renata Los ◽  
Anna Malm ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Fluorescence Microscopy ◽  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Bacterial Colonization ◽  
Formation Mechanisms ◽  
Microscopy Imaging ◽  
Vascular Prostheses ◽  
Coverage Ratio ◽  
Genetic Status

Objectives This study aims to examine biofilm formed on vascular prostheses by Staphylococcus epidermidis with different ica and aap genetic status, and to evaluate the effect of antibiotic-modified prostheses on bacterial colonization. Methods Biofilm formation was determined using fluorescence microscopy imaging. Quantitative analysis was conducted using the biofilm coverage ratio (BCR) calculations. Results Our investigations prove that the BCR method with fluorescent dye enabled an accurate assessment of biofilm coverage and comparison of the obtained results. The ica+ aap+ strains formed a biofilm on all of the examined vascular prostheses. Uni-Graft® modified with covalently immobilized amikacin was effective in preventing bacterial adherence. Conclusions Molecular biology techniques combined with phenotype studies give a broad insight into biofilm formation mechanisms. On the other hand, fluorescence microscopy imaging along with BCR calculations are reliable and simple tools to quantitatively estimate biofilm formation, as well as the effectiveness of antimicrobial prosthesis modification.

scholarly journals Phenotypic and Genetic Analysis of Biofilm Formation by Staphylococcus epidermidis

Medicina ◽  
2012 ◽  
Vol 48 (6) ◽  
pp. 45 ◽  
Author(s):  
Iveta Līduma ◽  
Tatjana Tračevska ◽  
Uģis Bērs ◽  
Aija Žileviča
Keyword(s):  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Clinical Isolates ◽  
Control Group ◽  
Clinical Group ◽  
Clinical Strains ◽  
Implanted Medical Devices ◽  
Important Virulence Factor ◽  
Accumulation Associated Protein

Objective. The most important virulence factor of Staphylococcus epidermidis is their capability to form a biofilm on the surfaces of implanted medical devices. The accumulative phase of biofilm formation is linked to the production of intercellular adhesin encoded by the icaADBC operon and accumulation-associated protein by the aap gene. The aim of the study was to investigate biofilm formation phenotypically and genetically in clinical strains of S. epidermidis in comparison with commensal strains. Material and Methods. The study was carried out in 4 hospitals in Riga, Latvia. In total, 105 clinical strains of Staphylococcus epidermidis isolated from patients’ blood (n=67) and intravenous catheters (n=38) in a case of laboratory-confirmed bacteremia were studied. Moreover, 60 Staphylococcus epidermidis commensal strains isolated from nose epithelium of healthy people were included as a control group. Appearance of the icaA and aap genes was tested by polymerase chain reaction. The microtiter plate method was used. Results. Biofilm formation was detected in 50 (47%) of Staphylococcus epidermidis isolates in the clinical group and 15 (25%) of isolates in the control group (P=0.0049). Among 50 biofilmforming clinical isolates, 46 (92%) were positive for the icaA and/or aap genes. The icaA and aap genes were not found only in 4 strains. Conclusions. The clinical isolates of Staphylococcus epidermidis were more likely to form biofilms than the commensal strains. The carriage of the icaA or aap gene alone, or their absence, is not applicable as a molecular marker for the discrimination invasive Staphylococcus epidermidis strains from contaminants.

scholarly journals Vitamin E for prevention of biofilm-caused Healthcare-associated infections

Open Medicine ◽  
2019 ◽  
Vol 15 (1) ◽  
pp. 14-21 ◽  
Author(s):  
Franca Vergalito ◽  
Laura Pietrangelo ◽  
Giulio Petronio Petronio ◽  
Federica Colitto ◽  
Marco Alfio Cutuli ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Vitamin E ◽  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Bacterial Colonization ◽  
Bacterial Biofilm ◽  
Tocopheryl Acetate ◽  
Healthcare Associated Infections ◽  
Biofilm Development ◽  
Healthcare Associated

AbstractThe healthcare-associated infections (HCAIs) occur in patients both in nosocomial environments and in community. More often HCAIs are associated to the use of medical devices and bacterial biofilm development on these equipments. Due to the clinical and economic relevance of this topic, new strategies for the treatment of infections caused by biofilm proliferation are unceasingly searched by scientists.The present study investigated the role of vitamin E to reduce the biofilm formation for a larger panel of human pathogens, including strains of Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Acinetobacter baumannii, Pseudomonas aeruginosa and Pseudomonas putida.This potential activity was tested by placing a preparation of vitamin E (α-Tocopheryl acetate) as interface between the bacterial culture and the polystyrene walls of a 96 well plate at different concentrations of glucose, used as a biofilm enhancer.The Staphylococcus genus was further investigated by spreading the vitamin E on a silicone catheter lumen and evaluating its influence on the bacterial colonization.From our results, vitamin E has been able to interfere with bacterial biofilm and prevent in vitro biofilm formation. Furthermore, the ability of Staphylococcus aureus and Staphylococcus epidermidis to colonize the catheter surface decreased as a result of vitamin E application.

scholarly journals The biofilm adhesion protein Aap from Staphylococcus epidermidis forms zinc-dependent amyloid fibers

2020 ◽  
Vol 295 (14) ◽  
pp. 4411-4427 ◽  
Author(s):  
Alexander E. Yarawsky ◽  
Stefanie L. Johns ◽  
Peter Schuck ◽  
Andrew B. Herr
Keyword(s):  
Staphylococcus Epidermidis ◽  
Biofilm Formation ◽  
Fiber Formation ◽  
Amyloid Fibers ◽  
Adhesion Protein ◽  
Amyloid Fiber Formation ◽  
Necessary And Sufficient ◽  
Hospital Acquired ◽  
Amyloid Assembly ◽  
Accumulation Associated Protein

The skin-colonizing commensal bacterium Staphylococcus epidermidis is a leading cause of hospital-acquired and device-related infections. Its pathogenicity in humans is largely due to its propensity to form biofilms, surface-adherent bacterial accumulations that are remarkably resistant to chemical and physical stresses. Accumulation-associated protein (Aap) from S. epidermidis has been shown to be necessary and sufficient for mature biofilm formation and catheter infection. Aap contains up to 17 tandem B-repeat domains, capable of zinc-dependent assembly into twisted, rope-like intercellular filaments in the biofilm. Using microscopic and biophysical techniques, we show here that Aap B-repeat constructs assemble further into zinc-dependent functional amyloid fibers. We observed such amyloid fibers by confocal microscopy during both early and late stages of S. epidermidis biofilm formation, and we confirmed that extracellular fibrils from these biofilms contain Aap. Unlike what has been observed for amyloidogenic biofilm proteins from other bacteria, which typically use chaperones or initiator proteins to initiate amyloid assembly, our findings indicate that Aap from S. epidermidis requires Zn2+ as a catalyst that drives amyloid fiber formation, similar to many mammalian amyloid-forming proteins that require metals for assembly. This work provides detailed insights into S. epidermidis biofilm formation and architecture that improve our understanding of persistent staphylococcal infections.

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