The Childrens Hospital of the Kings Daughters

Norfolk, United States

The Childrens Hospital of the Kings Daughters

Norfolk, United States
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Hair P.S.,Eastern Virginia Medical School | Foley C.K.,Eastern Virginia Medical School | Krishna N.K.,Eastern Virginia Medical School | Nyalwidhe J.O.,Eastern Virginia Medical School | And 5 more authors.
Results in Immunology | Year: 2013

Staphylococcus aureus is a premier human pathogen and the most common cause of osteoarticular, wound, and implanted device infections. We recently demonstrated S. aureus efficiently binds the classical complement regulator C4b-binding protein (C4BP) inhibiting antibody-initiated complement-mediated opsonization. Here we identify S. aureus surface protein SdrE as a C4BP-binding protein. Recombinant SdrE and recombinant bone sialoprotein-binding protein (Bbp), an allelic variant of SdrE, both efficiently bound to C4BP in heat-inactivated human serum. We previously described SdrE as binding alternative pathway regulator factor H. Recombinant SdrE and Bbp efficiently bound C4BP and factor H in serum without apparent interference. Gain of function studies utilizing Lactococcus lactis clones expressing SdrE or Bbp increased serum C4BP and factor H binding, compared with empty-vector control (WT) approximately 2-fold. Correspondingly, classical pathway-mediated C3-fragment opsonization and bacterial killing by human neutrophils decreased by half for L. lactis clones expressing SdrE or Bbp compared with WT. In summary, we identify SdrE and allelic variant Bbp as S. aureus surface proteins that bind the complement regulator C4BP inhibiting classical pathway-mediated bacterial opsonization and killing. © 2013 The Authors.

Hair P.S.,Eastern Virginia Medical School | Echague C.G.,Eastern Virginia Medical School | Rohn R.D.,Eastern Virginia Medical School | Rohn R.D.,Childrens Specialty Group | And 7 more authors.
Journal of Translational Medicine | Year: 2012

Background: Diabetic patients are at increased risk for bacterial infections; these studies provide new insight into the role of the host defense complement system in controlling bacterial pathogens in hyperglycemic environments.Methods: The interactions of complement C3 with bacteria in elevated glucose were assayed for complement activation to opsonic forms, phagocytosis and bacterial killing. C3 was analyzed in euglycemic and hyperglycemic conditions by mass spectrometry to measure glycation and structural differences.Results: Elevated glucose inhibited S. aureus activation of C3 and deposition of C3b and iC3b on the bacterial surface. S. aureus-generated C5a and serum-mediated phagocytosis by neutrophils were both decreased in elevated glucose conditions. Interestingly, elevated glucose increased the binding of unactivated C3 to S. aureus, which was reversible on return to normal glucose concentrations. In a model of polymicrobial infection, S. aureus in elevated glucose conditions depleted C3 from serum resulting in decreased complement-mediated killing of E. coli. To investigate the effect of differing glucose concentration on C3 structure and glycation, purified C3 incubated with varying glucose concentrations was analyzed by mass spectrometry. Glycation was limited to the same three lysine residues in both euglycemic and hyperglycemic conditions over one hour, thus glycation could not account for observed changes between glucose conditions. However, surface labeling of C3 with sulfo-NHS-biotin showed significant changes in the surface availability of seven lysine residues in response to increasing glucose concentrations. These results suggest that the tertiary structure of C3 changes in response to hyperglycemic conditions leading to an altered interaction of C3 with bacterial pathogens.Conclusions: These results demonstrate that hyperglycemic conditions inhibit C3-mediated complement effectors important in the immunological control of S. aureus. Mass spectrometric analysis reveals that the glycation state of C3 is the same regardless of glucose concentration over a one-hour time period. However, in conditions of elevated glucose C3 appears to undergo structural changes. © 2012 Hair et al; licensee BioMed Central Ltd.

Hair P.S.,Eastern Virginia Medical School | Wagner S.M.,Eastern Virginia Medical School | Friederich P.T.,Eastern Virginia Medical School | Drake R.R.,Eastern Virginia Medical School | And 4 more authors.
Molecular Immunology | Year: 2012

Staphylococcus aureus is the major cause of human skin and soft-tissue infections as well as invasive infections like post-operative wound infections, septic arthritis, and osteomyelitis. The complement system plays an important role in the immunological control of many bacteria, but can be inhibited by a variety of strategies including recruitment of complement regulatory proteins like C4b-binding protein (C4BP). These experiments demonstrate that S. aureus opsonization with C4b occurs rapidly in serum and is predominantly initiated by anti-staphylococcal antibodies. Much of the S. aureus-bound C4b is quickly cleaved to the inactive forms iC4b and C4d. Clinical S. aureus strains rapidly bind significant amounts of the complement regulator C4BP from serum. S. aureus also binds purified C4BP. S. aureus-bound C4BP functions as a cofactor for factor I-mediated C4b cleavage to iC4b and C4d. In the absence of factor I, C4BP decreases classical pathway-mediated deposition of C3b on the S. aureus surface by inhibiting the classical pathway C3-convertase. In summary, C4BP is recruited to the S. aureus surface where it functions to inhibit C4 complement effectors, suggesting a previously undescribed immune evasion strategy for this pathogen. © 2012 Elsevier Ltd.

Yetisen A.K.,University of Cambridge | Butt H.,University of Cambridge | da Cruz Vasconcellos F.,University of Cambridge | Montelongo Y.,University of Cambridge | And 9 more authors.
Advanced Optical Materials | Year: 2014

This tunable holographic sensor offers interrogation and a reporting transducer as well as an analyte-responsive hydrogel, rendering it label-free and reusable. A single 6 ns laser pulse is used to fabricate holographic sensors consisting of silver nanoparticles arranged periodically within a polymer film. The tunability of the sensor is demonstrated through pH sensing of artificial urine and validated through computational modeling. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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