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Proteomic analysis reveals pathogen-derived biomarkers of acute babesiosis in erythrocytes, plasma, and urine of infected hamsters

  • Immunology and Host-Parasite Interactions - Original Paper
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Abstract

Babesiosis among humans is on the rise in North America. Current diagnostic assays for the screening of babesiosis require blood collection by venipuncture, which is an invasive method. Urine on the other hand is a desirable biospecimen for biomarker analysis of Babesia microti infections because it can be collected periodically and non-invasively. Our group uses a new class of biomarker harvesting nanocage technology, which, when combined with mass spectrometry (MS), can determine the presence of parasite proteins shed in different bodily fluids of mammalian hosts, including urine. Using the hamster model of babesiosis, our nanoparticle-MS approach identified several B. microti proteins in erythrocytes, plasma, and urine samples. Surface and secreted antigens previously shown to elicit host immune responses against the parasite were particularly abundant in erythrocytes and plasma compared to other proteins. Two of these antigens, BmSA1 and BMR1_03g00947, showed different localization patterns by immunofluorescence of infected erythrocytes. Hamster urine samples from parasitemic animals harbored lower numbers of B. microti proteins compared to erythrocytes and plasma, with glycolytic enzymes, cytoskeletal components, and chaperones being the most frequently detected proteins. By applying novel nanoparticle-MS methods, a high level of analytical sensitivity can be achieved to detect multiple B. microti proteins in blood and urine. This is generally difficult to obtain with other techniques due to the masking of parasite biomarkers by the complex biomolecular matrix of bodily fluids from the host.

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Acknowledgments

We thank Debra Adams-Fish from the ATCC® Specialty Laboratory Facilities for her assistance in the animal experiments. ATCC® is a registered trademark of the American Type Culture Collection.

Funding

R.E. Molestina was supported by the ATCC® Internal Research and Development Program. A. Luchini, L. Liotta, and R. Magni were supported by grants W81XWH-17-1-0175 from the US Army Medical Research Acquisition Activity, and R21HD097472, R01AR068436, and R21AI138135 from the US National Institutes of Health.

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Contributions

R.E. Molestina contributed to the study conception and design, animal experiments, biological material preparation, results figures, and first draft of the manuscript. Nanoparticle synthesis, protein capture, MS analysis, and data collection were performed by R. Magni. All authors commented on previous versions of the manuscript, provided edits, and read and approved the final manuscript.

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Correspondence to Robert E. Molestina.

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Section Editor: Julia Walochnik

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Magni, R., Luchini, A., Liotta, L. et al. Proteomic analysis reveals pathogen-derived biomarkers of acute babesiosis in erythrocytes, plasma, and urine of infected hamsters. Parasitol Res 119, 2227–2235 (2020). https://doi.org/10.1007/s00436-020-06712-5

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  • DOI: https://doi.org/10.1007/s00436-020-06712-5

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