Monoclonal Antibodies against Accumulation-Associated Protein Affect EPS Biosynthesis and Enhance Bacterial Accumulation of Staphylococcus epidermidis

Hu, Jianguo; Xu, Tao; Zhu, Tao; Lou, Qiang; Wang, Xueqin; Wu, Yang; Huang, Ruoyu; Liu, Jingran; Liu, Huayong; Yu, Fangyou; Ding, Baixing; Huang, Ya-Lin; Tong, Wenyan; Qu, Di

doi:10.1371/journal.pone.0020918

Cited by 41 publications

(31 citation statements)

References 41 publications

(76 reference statements)

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“…3 and 4) and SesC (Fig. 4); this phenomenon has been documented previously for Aap (16). Antibodies to EmbP performed well in the prophylactic assay (Fig.…”

Section: Discussionsupporting

confidence: 84%

“…Proteins were expressed in Escherichia coli BL21(DE3) using Overnight Express Autoinduction System 1 (EMD Millipore), per the manufacturer's protocol. For Aap/SERP2398, a 20-amino-acid peptide (PGKPGVK NPDTGEVVTPPVD) was synthesized for immunization (GenScript), based on its published identification as an antibiofilm epitope (16). Expression cultures were harvested by centrifugation, and cell pellets were stored at Ϫ80°C.…”

Section: Methodsmentioning

confidence: 99%

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Antibodies to PhnD Inhibit Staphylococcal Biofilms

et al. 2014

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Biofilm formation on central lines or peripheral catheters is a serious threat to patient well-being. Contaminated vascular devices can act as a nidus for bloodstream infection and systemic pathogen dissemination. Staphylococcal biofilms are the most common cause of central-line-associated bloodstream infections, and antibiotic resistance makes them difficult to treat. As an alternative to antibiotic intervention, we sought to identify anti-staphylococcal biofilm targets for the development of a vaccine or antibody prophylactic. A screening strategy was devised using a microfluidic system to test antibody-mediated biofilm inhibition under biologically relevant conditions of shear flow. Affinity-purified polyclonal antibodies to target antigen PhnD inhibited both Staphylococcus epidermidis and S. aureus biofilms. PhnD-specific antibodies blocked biofilm development at the initial attachment and aggregation stages, and deletion of phnD inhibited normal biofilm formation. We further adapted our microfluidic biofilm system to monitor the interaction of human neutrophils with staphylococcal biofilms and demonstrated that PhnD-specific antibodies also serve as opsonins to enhance neutrophil binding, motility, and biofilm engulfment. These data support the identification of PhnD as a lead target for biofilm intervention strategies performed either by vaccination or through passive administration of antibodies.

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“…3 and 4) and SesC (Fig. 4); this phenomenon has been documented previously for Aap (16). Antibodies to EmbP performed well in the prophylactic assay (Fig.…”

Section: Discussionsupporting

confidence: 84%

Section: Methodsmentioning

confidence: 99%

Antibodies to PhnD Inhibit Staphylococcal Biofilms

et al. 2014

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“…Alternatively, the rod-like structure might play a role in maintaining bacterial separation in the biofilm. Furthermore, a recent study on Aapmediated biofilm formation by S. epidermidis showed that anti-G5 monoclonal antibodies enhanced bacterial accumulation, whereas those with an epitope in the E segment inhibited biofilm accumulation to 60% of the maximum (42). This suggests that G5 and E domains might play different roles during bacterial accumulation, despite their structural similarity.…”

Section: Discussionmentioning

confidence: 98%

Staphylococcal biofilm-forming protein has a contiguous rod-like structure

Gruszka

Wojdyla

Bingham

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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Staphylococcus aureus and Staphylococcus epidermidis form communities (called biofilms) on inserted medical devices, leading to infections that affect many millions of patients worldwide and cause substantial morbidity and mortality. As biofilms are resistant to antibiotics, device removal is often required to resolve the infection. Thus, there is a need for new therapeutic strategies and molecular data that might assist their development. Surface proteins S. aureus surface protein G (SasG) and accumulation-associated protein ( S. epidermidis ) promote biofilm formation through their “B” regions. B regions contain tandemly arrayed G5 domains interspersed with approximately 50 residue sequences (herein called E) and have been proposed to mediate intercellular accumulation through Zn 2+ -mediated homodimerization. Although E regions are predicted to be unstructured, SasG and accumulation-associated protein form extended fibrils on the bacterial surface. Here we report structures of E–G5 and G5–E–G5 from SasG and biophysical characteristics of single and multidomain fragments. E sequences fold cooperatively and form interlocking interfaces with G5 domains in a head-to-tail fashion, resulting in a contiguous, elongated, monomeric structure. E and G5 domains lack a compact hydrophobic core, and yet G5 domain and multidomain constructs have thermodynamic stabilities only slightly lower than globular proteins of similar size. Zn 2+ does not cause SasG domains to form dimers. The work reveals a paradigm for formation of fibrils on the 100-nm scale and suggests that biofilm accumulation occurs through a mechanism distinct from the “zinc zipper.” Finally, formation of two domains by each repeat (as in SasG) might reduce misfolding in proteins when the tandem arrangement of highly similar sequences is advantageous.

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“…Previous investigations have demonstrated that Aap domain architecture and subsequent processing is variable among clinical and laboratory strains (12,26,33,35,39). Draft genomic sequencing revealed that Aap of 1457 has an A domain with 11 short, 16-aa repeats, followed by the conserved L-type lectin domain, while the B domain contains 6 full and 1 partial B repeat followed by 2 putative collagen-like repeat motifs.…”

Section: S Epidermidis Strain 1457 Produces and Processes Aap In Vitromentioning

confidence: 99%

“…Clearly, additional molecules are capable of supporting biofilms in vivo. Among these, Aap is the most well studied and frequently identified component of PIA-independent, protein-mediated S. epidermidis biofilms (12,(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43). Aap is a cell wall-anchored protein (41) of approximately 220 kDa; however, the size varies between strains due to differences in the number of B domain repeats (Fig.…”

mentioning

confidence: 99%

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

et al. 2015

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Biofilm formation is the primary virulence factor of Staphylococcus epidermidis. S. epidermidis biofilms 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 an aap allelic replacement mutant and an icaADBC aap double mutant. When subjected to fluid shear, strains deficient in Aap production produced significantly less biofilm than Aap-positive strains. To examine the in vivo relevance 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 the aap mutant with full-length Aap (containing the A domain), but not the B domain alone, increased initial attachment to microtiter plates, as did in trans expression of the A domain in adhesion-deficient Staphylococcus carnosus. These results demonstrate Aap contributes to S. epidermidis infection, which may in part be due to A domain-mediated attachment to abiotic surfaces.

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Monoclonal Antibodies against Accumulation-Associated Protein Affect EPS Biosynthesis and Enhance Bacterial Accumulation of Staphylococcus epidermidis

Cited by 41 publications

References 41 publications

Antibodies to PhnD Inhibit Staphylococcal Biofilms

Antibodies to PhnD Inhibit Staphylococcal Biofilms

Staphylococcal biofilm-forming protein has a contiguous rod-like structure

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

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