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Bacterial Infections

Tracking Anti-Staphylococcus aureus Antibodies Produced In Vivo and Ex Vivo during Foot Salvage Therapy for Diabetic Foot Infections Reveals Prognostic Insights and Evidence of Diversified Humoral Immunity

Irvin Oh, Gowrishankar Muthukrishnan, Mark J. Ninomiya, James D. Brodell Jr., Benjamin L. Smith, Charles C. Lee, Steven R. Gill, Christopher A. Beck, Edward M. Schwarz, John L. Daiss
Victor J. Torres, Editor
Irvin Oh
aCenter for Musculoskeletal Research and Department of Orthopaedics, University of Rochester Medical Center, Rochester, New York, USA
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Gowrishankar Muthukrishnan
aCenter for Musculoskeletal Research and Department of Orthopaedics, University of Rochester Medical Center, Rochester, New York, USA
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  • ORCID record for Gowrishankar Muthukrishnan
Mark J. Ninomiya
aCenter for Musculoskeletal Research and Department of Orthopaedics, University of Rochester Medical Center, Rochester, New York, USA
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James D. Brodell Jr.
aCenter for Musculoskeletal Research and Department of Orthopaedics, University of Rochester Medical Center, Rochester, New York, USA
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Benjamin L. Smith
aCenter for Musculoskeletal Research and Department of Orthopaedics, University of Rochester Medical Center, Rochester, New York, USA
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Charles C. Lee
aCenter for Musculoskeletal Research and Department of Orthopaedics, University of Rochester Medical Center, Rochester, New York, USA
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Steven R. Gill
bDepartment of Microbiology and Immunology, School of Medicine and Dentistry, University of Rochester, Rochester, New York, USA
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Christopher A. Beck
aCenter for Musculoskeletal Research and Department of Orthopaedics, University of Rochester Medical Center, Rochester, New York, USA
cDepartment of Biostatistics and Computational Biology, University of Rochester, Rochester, New York, USA
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Edward M. Schwarz
aCenter for Musculoskeletal Research and Department of Orthopaedics, University of Rochester Medical Center, Rochester, New York, USA
bDepartment of Microbiology and Immunology, School of Medicine and Dentistry, University of Rochester, Rochester, New York, USA
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John L. Daiss
aCenter for Musculoskeletal Research and Department of Orthopaedics, University of Rochester Medical Center, Rochester, New York, USA
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Victor J. Torres
New York University School of Medicine
Roles: Editor
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DOI: 10.1128/IAI.00629-18
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ABSTRACT

Management of foot salvage therapy (FST) for diabetic foot infections (DFI) is challenging due to the absence of reliable diagnostics to identify the etiologic agent and prognostics to justify aggressive treatments. As Staphylococcus aureus is the most common pathogen associated with DFI, we aimed to develop a multiplex immunoassay of IgG in serum and medium enriched for newly synthesized anti-S. aureus antibodies (MENSA) generated from cultured peripheral blood mononuclear cells of DFI patients undergoing FST. Wound samples were collected from 26 DFI patients to identify the infecting bacterial species via 16S rRNA sequencing. Blood was obtained over 12 weeks of FST to assess anti-S. aureus IgG levels in sera and MENSA. The results showed that 17 out of 26 infections were polymicrobial and 12 were positive for S. aureus. While antibody titers in serum and MENSA displayed similar diagnostic potentials to detect S. aureus infection, MENSA showed a 2-fold-greater signal-to-background ratio. Multivariate analyses revealed increases in predictive power of diagnosing S. aureus infections (area under the receiver operating characteristic curve [AUC] > 0.85) only when combining titers against different classes of antigens, suggesting cross-functional antigenic diversity. Anti-S. aureus IgG levels in MENSA decreased with successful FST and rose with reinfection. In contrast, IgG levels in serum remained unchanged throughout the 12-week FST. Collectively, these results demonstrate the applicability of serum and MENSA for diagnosis of S. aureus DFI with increased power by combining functionally distinct titers. We also found that tracking MENSA has prognostic potential to guide clinical decisions during FST.

FOOTNOTES

    • Received 12 August 2018.
    • Returned for modification 29 August 2018.
    • Accepted 20 September 2018.
    • Accepted manuscript posted online 1 October 2018.
  • Supplemental material for this article may be found at https://doi.org/10.1128/IAI.00629-18.

  • Copyright © 2018 American Society for Microbiology.

All Rights Reserved.

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Tracking Anti-Staphylococcus aureus Antibodies Produced In Vivo and Ex Vivo during Foot Salvage Therapy for Diabetic Foot Infections Reveals Prognostic Insights and Evidence of Diversified Humoral Immunity
Irvin Oh, Gowrishankar Muthukrishnan, Mark J. Ninomiya, James D. Brodell Jr., Benjamin L. Smith, Charles C. Lee, Steven R. Gill, Christopher A. Beck, Edward M. Schwarz, John L. Daiss
Infection and Immunity Nov 2018, 86 (12) e00629-18; DOI: 10.1128/IAI.00629-18

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Tracking Anti-Staphylococcus aureus Antibodies Produced In Vivo and Ex Vivo during Foot Salvage Therapy for Diabetic Foot Infections Reveals Prognostic Insights and Evidence of Diversified Humoral Immunity
Irvin Oh, Gowrishankar Muthukrishnan, Mark J. Ninomiya, James D. Brodell Jr., Benjamin L. Smith, Charles C. Lee, Steven R. Gill, Christopher A. Beck, Edward M. Schwarz, John L. Daiss
Infection and Immunity Nov 2018, 86 (12) e00629-18; DOI: 10.1128/IAI.00629-18
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KEYWORDS

Staphylococcus aureus
diabetes
diabetic foot infections
diagnostics
immunoassays
plasmablasts

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