Immunoproteomic analysis of Borrelia miyamotoi for the identification of serodiagnostic antigens

• Published in: Scientific reports 2019-11, Vol.9 (1), p.16808-14, Article 16808
• Main Authors: Harris, Emma K., Harton, Marisa R., de Mello Marques, Maria Angela, Belisle, John T., Molins, Claudia R., Breuner, Nicole, Wormser, Gary P., Gilmore, Robert D.
• Format: Article
• Language: English
• Subjects: 38/22; 631/326/1320; 631/326/2521; 82/1; 82/29; 82/58; 82/80; 82/83; Animals; Antibodies, Protozoan - metabolism; Antigens; Antigens, Protozoan - immunology; Borrelia - immunology; Borrelia Infections - immunology; Borrelia miyamotoi; Electrophoresis, Gel, Two-Dimensional; Female; Humanities and Social Sciences; Humans; Immune response; Immunoblotting; Immunogenicity; Immunoglobulin G; Inoculation; Insect bites; Lipoproteins; Lipoproteins - immunology; Lyme disease; Mass Spectrometry; Mass spectroscopy; Mice; multidisciplinary; Proteins; Proteomics - methods; Public health; Science; Science (multidisciplinary); Serologic Tests; Spirochetes; Tick Bites - immunology; Tick Bites - parasitology; Vector-borne diseases
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-53248-5

The tick-borne spirochete, Borrelia miyamotoi , is an emerging pathogen of public health significance. Current B . miyamotoi serodiagnostic testing depends on reactivity against GlpQ which is not highly sensitive on acute phase serum samples. Additionally, anti- B . miyamotoi antibodies can cross-react with C6 antigen testing for B . burgdorferi , the causative agent of Lyme disease, underscoring the need for improved serological assays that produce accurate diagnostic results. We performed an immunoproteomics analysis of B . miyamotoi proteins to identify novel serodiagnostic antigens. Sera from mice infected with B . miyamotoi by subcutaneous inoculation or tick bite were collected for immunoblotting against B . miyamotoi membrane-associated proteins separated by 2-dimensional electrophoresis (2DE). In total, 88 proteins in 40 2DE immunoreactive spots were identified via mass spectrometry. Multiple variable large proteins (Vlps) and a putative lipoprotein were among those identified and analyzed. Reactivity of anti- B . miyamotoi sera against recombinant Vlps and the putative lipoprotein confirmed their immunogenicity. Mouse anti- B . burgdorferi serum was cross-reactive to all recombinant Vlps, but not against the putative lipoprotein by IgG. Furthermore, antibodies against the recombinant putative lipoprotein were present in serum from a B . miyamotoi -infected human patient, but not a Lyme disease patient. Results presented here provide a comprehensive profile of B . miyamotoi antigens that induce the host immune response and identify a putative lipoprotein as a potentially specific antigen for B . miyamotoi serodetection.