Title: Serum biomarkers of Burkholderia mallei infection elucidated by proteomic imaging of skin and lung abscesses

Abstract

Background: The bacterium Burkholderia mallei is the etiological agent of glanders, a highly contagious, often fatal zoonotic infectious disease that is also a biodefense concern. Clinical laboratory assays that analyze blood or other biological fluids are the highest priority because these specimens can be collected with minimal risk to the patient. However, progress in developing sensitive assays for monitoring B. mallei infection is hampered by a shortage of useful biomarkers. Results: Reasoning that there should be a strong correlation between the proteomes of infected tissues and circulating serum, we employed imaging mass spectrometry (IMS) of thin-sectioned tissues from Chlorocebus aethiops (African green) monkeys infected with B. mallei to localize host and pathogen proteins that were associated with abscesses. Using laser-capture microdissection of specific regions identified by IMS and histology within the tissue sections, a more extensive proteomic analysis was performed by a technique that combined the physical separation capabilities of liquid chromatography (LC) with the sensitive mass analysis capabilities of mass spectrometry (LC-MS/MS). By examining standard formalin-fixed, paraffin-embedded tissue sections, this strategy resulted in the identification of several proteins that were associated with lung and skin abscesses, including the host protein calprotectin and the pathogen protein GroEL. Elevated levels ofmore » calprotectin detected by ELISA and antibody responses to GroEL, measured by a microarray of the bacterial proteome, were subsequently detected in the sera of C. aethiops, Macaca mulatta, and Macaca fascicularis primates infected with B. mallei. Conclusions: Our results demonstrate that a combination of multidimensional MS analysis of traditional histology specimens with high-content protein microarrays can be used to discover lead pairs of host-pathogen biomarkers of infection that are identifiable in biological fluids.« less

Authors:

Glaros, Trevor G. ^[1]; Blancett, Candace D. ^[2]; Bell, Todd M. ^[2]; Natesan, Mohan ^[1]; Ulrich, Robert G. ^[1]

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+ Show Author Affiliations

1. US Army Medical Research Inst. of Infectious Diseases (USAMRIID), Frederick, MD (United States). Molecular and Translational Sciences
2. US Army Medical Research Inst. of Infectious Diseases (USAMRIID), Frederick, MD (United States). Pathology

Publication Date:

Tue Mar 10 00:00:00 EDT 2015

Research Org.:

Oak Ridge Institute for Science and Education (ORISE), Oak Ridge, TN (United States)

Sponsoring Org.:

USDOE Office of Science (SC); Defense Threat Reduction Agency (DTRA)

OSTI Identifier:

1626750

Grant/Contract Number:  

SC0010687; CB3498

Resource Type:

Accepted Manuscript

Journal Name:

Clinical Proteomics

Additional Journal Information:

Journal Volume: 12; Journal Issue: 1; Journal ID: ISSN 1542-6416

Publisher:

BioMed Central

Country of Publication:

United States

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES; Biochemistry & Molecular Biology; Imaging mass spectrometry; Biomarker; Burkholderia mallei; Burkholderia pseudomallei; Laser capture microdissection; LC-MS/MS; Protein microarray; Glanders; Melioidosis; GroEL; Calprotectin; Formalin-fixed paraffin embedded tissue; FFPE