Proteomic analysis of threeBorrelia burgdorferisensu lato native species and disseminating clones: Relevance for Lyme vaccine design

Abstract

Lyme borreliosis is the most important vector-borne disease in the Northern hemisphere. It is caused by Borrelia burgdorferi sensu lato bacteria transmitted to humans by the bite of hard ticks, Ixodes spp. Although antibiotic treatments are efficient in the early stage of the infection, a significant number of patients develop disseminated manifestations (articular, neurological, and cutaneous) due to unnoticed or absence of erythema migrans, or to inappropriate treatment. Vaccine could be an efficient approach to decrease Lyme disease incidence. We have developed a proteomic approach based on a one dimensional gel electrophoresis followed by LC-MS/MS strategy to identify new vaccine candidates. We analyzed a disseminating clone and the associated wild-type strain for each major pathogenic Borrelia species: B. burgdorferi sensu stricto, B. garinii, and B. afzelii. We identified specific proteins and common proteins to the disseminating clones of the three main species. In parallel, we used a spectral counting strategy to identify upregulated proteins common to the clones. Finally, 40 proteins were found that could potentially be involved in bacterial virulence and of interest in the development of a new vaccine. We selected the three proteins specifically detected in the disseminating clones of the three Borrelia species and checked by RT-PCR whether they are expressed in mouse skin upon B. burgdorferi ss inoculation. Interestingly, BB0566 appears as a potential vaccine candidate. All MS data have been deposited in the ProteomeXchange with identifier PXD000876 (http://proteomecentral.proteomexchange.org/dataset/PXD000876).

Heterogeneity of Borrelia burgdorferi Sensu Stricto Population and Its Involvement in Borrelia Pathogenicity: Study on Murine Model with Specific Emphasis on the Skin Interface

“…A total of 887 proteins were identified with 848 proteins in 297/4 and 777 proteins in the parental strain ( Fig 5A and S2 Table ). As compared to our previous results [ 18 ], we observed a reproducibility of 89% of the total proteins identified. Interestingly, in this study, 110 proteins were detected specifically in the clone 297/4.…”

“…burgdorferi ss strain 297 [ 17 ]. We selected the clone 297/4 because of its cerebral dissemination at day 30 [ 18 ]. In addition, this clone induced neurological manifestations in a few mice (alteration of behavior and hemorraghes—data not shown).…”

“…burgdorferi 297 and the clone 297/4. We used a GeLC-MS/MS based on SDS-PAGE, systematic excision of the protein bands, and identification through nanoLC-MS/MS as described [ 18 ]. This analytical strategy does not specifically target membrane proteins but affords an exhaustive identification of B .…”

“…burgdorferi 297, since this strain is particularly disseminating and is defined as RST2 (intermediate in dissemination). We chose clone 4 for its cerebral localization compared to the others we isolated [ 18 ]. A recent study in horses also revealed the cerebral localization of a strain of B .…”

Proteomic Analyses of Morphological Variants ofBorrelia burgdorferiShed New Light on Persistence Mechanisms: Implications for Pathogenesis, Diagnosis and Treatment

“…Proteomics approach has been used previously to study the B. burgdorferi proteome difference during transmission between ticks and mammals [27], in response to environmental variations of temperature and pH [28], and between various native species [29]. However, previous studies provided little quantitative information, and did not compare the proteomes of morphological variants of B. burgdorferi in different growth period.…”