Discovery and Targeted Proteomics on Cutaneous Biopsies Infected by Borrelia to Investigate Lyme Disease*

Schnell, Gilles; Bœuf, Amandine; Westermann, Benoît; Jaulhac, B.; Lipsker, Dan; Carapito, Christine; Boulanger, Nathalie; Ehret‐Sabatier, Laurence

doi:10.1074/mcp.m114.046540

Cited by 20 publications

(32 citation statements)

References 60 publications

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“…The selection of a more specific body part such as gut and/or the use of other mass spectrometry strategies (i.e. targeted proteomics like Selected Reaction Monitoring in tandem with mass spectrometry [ 39 ]) could improve concomitant tick identification and/or pathogen detection [ 40 ]. However, the great reproducibility of the MS spectra generated from the three compartments would allow an identification using each one of these compartments.…”

Section: Discussionmentioning

confidence: 99%

Assessment of MALDI-TOF MS biotyping for Borrelia burgdorferi sl detection in Ixodes ricinus

et al. 2017

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Matrix Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) has been demonstrated to be useful for tick identification at the species level. More recently, this tool has been successfully applied for the detection of bacterial pathogens directly in tick vectors. The present work has assessed the detection of Borrelia burgdorferi sensu lato in Ixodes ricinus tick vector by MALDI-TOF MS. To this aim, experimental infection model of I. ricinus ticks by B. afzelii was carried out and specimens collected in the field were also included in the study. Borrelia infectious status of I. ricinus ticks was molecularly controlled using half-idiosome to classify specimens. Among the 39 ticks engorged on infected mice, 14 were confirmed to be infected by B. afzelii. For field collection, 14.8% (n = 12/81) I. ricinus ticks were validated molecularly as infected by B. burgdorferi sl. To determine the body part allowing the detection of MS protein profile changes between non-infected and B. afzelii infected specimens, ticks were dissected in three compartments (i.e. 4 legs, capitulum and half-idiosome) prior to MS analysis. Highly reproducible MS spectra were obtained for I. ricinus ticks according to the compartment tested and their infectious status. However, no MS profile change was found when paired body part comparison between non-infected and B. afzelii infected specimens was made. Statistical analyses did not succeed to discover, per body part, specific MS peaks distinguishing Borrelia-infected from non-infected ticks whatever their origins, laboratory reared or field collected. Despite the unsuccessful of MALDI-TOF MS to classify tick specimens according to their B. afzelii infectious status, this proteomic tool remains a promising method for rapid, economic and accurate identification of tick species. Moreover, the singularity of MS spectra between legs and half-idiosome of I. ricinus could be used to reinforce this proteomic identification by submission of both these compartments to MS.

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Section: Discussionmentioning

confidence: 99%

Assessment of MALDI-TOF MS biotyping for Borrelia burgdorferi sl detection in Ixodes ricinus

et al. 2017

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“…Although not observed in our two-dimensional gel, three more B. burgdorferi proteins, namely outer surface protein C (ospC), flagellar filament 41 kDa core protein (fla), and DNA-binding protein HU (hup) have also attracted our attention. These proteins were previously reported as abundant B. burgdorferi proteins 16, 23–26 and have unique tryptic peptides for selective detection of B. burgdorferi. Consequently, the list of tryptic peptides for the seven B. burgdorferi proteins was generated in silico .…”

Section: Resultsmentioning

confidence: 94%

“…Current instrumentation allows for the measurement of many proteins in a single sample, making MRM an ideal assay to perform high-throughput measurements on a panel of target proteins. 13–15 Successful application of the MRM assay for detection of Borrelia proteins in human skin biopsies has been recently reported, 16 however direct MRM assay in human blood or serum for early detection of Lyme disease poses additional challenges due to the extremely low abundance of total circulating Borrelia proteins.…”

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confidence: 99%

Quantification of Borrelia burgdorferi Membrane Proteins in Human Serum: A New Concept for Detection of Bacterial Infection

et al. 2015

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The B. burgdorferi spirochete is the causative agent of Lyme disease, the most common tick-borne disease in the United States. The low abundance of bacterial proteins in human serum during infection imposes a challenge for early proteomic detection of Lyme disease. To address this challenge, we propose to detect membrane proteins released from bacteria due to disruption of their plasma membrane triggered by the innate immune system. These membrane proteins can be separated from the bulk of serum proteins by high-speed centrifugation causing substantial sample enrichment prior to targeted protein quantification using multiple reaction monitoring mass spectrometry. This new approach was first applied to detection of B. burgdorferi membrane proteins supplemented in human serum. Our results indicated that detection of B. burgdorferi membrane proteins, which are ≈ 107 lower in abundance than major serum proteins, is feasible. Therefore quantitative analysis was also carried out for serum samples from three patients with acute Lyme disease. We were able to demonstrate the detection of ospA, the major B. burgdorferi lipoprotein at the level of 4.0 fmol of ospA/mg of serum protein. The results confirm the concept and suggest that the proposed approach can be expanded to detect other bacterial infections in humans, particularly where existing diagnostics are unreliable.

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“…[104] In addition, the solubilizing and denaturing capacity of SDS allows for extraction of membrane proteins. GeLC-SRM was also successfully applied to human skin biopsies,[117] serum/plasma[118] and liver tissue samples. [119] Nevertheless, this approach may not be suitable for some post-translationally modified proteins with a broad molecular weight range; the enrichment efficiency may also vary significantly for different proteins.…”

Section: Targeted Proteomics Approaches For Preclinical Verification mentioning

confidence: 99%

The clinical impact of recent advances in LC-MS for cancer biomarker discovery and verification

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Shi

Qian

et al. 2015

Expert Review of Proteomics

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Mass spectrometry (MS) -based proteomics has become an indispensable tool with broad applications in systems biology and biomedical research. With recent advances in liquid chromatography (LC) and MS instrumentation, LC–MS is making increasingly significant contributions to clinical applications, especially in the area of cancer biomarker discovery and verification. To overcome challenges associated with analyses of clinical samples (for example, a wide dynamic range of protein concentrations in bodily fluids and the need to perform high throughput and accurate quantification of candidate biomarker proteins), significant efforts have been devoted to improve the overall performance of LC–MS-based clinical proteomics platforms. Reviewed here are the recent advances in LC–MS and its applications in cancer biomarker discovery and quantification, along with the potentials, limitations and future perspectives.

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Discovery and Targeted Proteomics on Cutaneous Biopsies Infected by Borrelia to Investigate Lyme Disease*

Cited by 20 publications

References 60 publications

Assessment of MALDI-TOF MS biotyping for Borrelia burgdorferi sl detection in Ixodes ricinus

Assessment of MALDI-TOF MS biotyping for Borrelia burgdorferi sl detection in Ixodes ricinus

Quantification of Borrelia burgdorferi Membrane Proteins in Human Serum: A New Concept for Detection of Bacterial Infection

The clinical impact of recent advances in LC-MS for cancer biomarker discovery and verification

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