Identification of Borrelia protein candidates in mouse skin for potential diagnosis of disseminated Lyme borreliosis

Grillon, Antoine; Westermann, BenoÃ®t.; Cantero, Paola; Jaulhac, B.; Voordouw, Maarten J.; Kapps, Delphine; Collin, Elody; Barthel, Cathy; Ehret‐Sabatier, Laurence; Boulanger, Nathalie

doi:10.1038/s41598-017-16749-9

Cited by 21 publications

(35 citation statements)

References 66 publications

(76 reference statements)

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“…The puzzling results of these studies with enormous inocula were also contested by other authors [70], since the maximal concentration of LB spirochetes achievable in suitable culture medium is ~ 10 8 -10 9 per ml [71,72]. In comparison, only a small number of spirochetes per injection site (up to 10 3 organisms) is deposited into the dermis of mice after tick attachment [61,[73][74][75][76]. Hence, not only the strict IV inoculation but also a reasonable inoculum dose plays the most crucial role in establishing a reliable murine model and drawing a realistic conclusion.…”

Section: Discussionmentioning

confidence: 91%

“…The lack of Bbss-specific antibodies in the course of the intravenous injection strongly suggests that these spirochetes did not even multiply or provide sufficient antigen to stimulate the host B cell population. However, LB spirochetes inoculated into the skin can persist and multiply locally and disseminate to distal sites [61]. During dissemination, the blood seems to play no role because Bbss organisms were not detected in any blood sample.…”

Section: Discussionmentioning

confidence: 99%

“…They and others [63] have also highlighted the issue that prompt excision or topical treatment with antibiotics applied in the site of murine skin shortly after tick exposure or needle inoculation can avert systemic infection. Moreover, immunosuppression with dermocorticoid clobetasol reactivated LB Borrelia abundance in the skin tissue, while the blood still remained spirochete negative [61]. In contrast, high levels of spirochetemia were achieved in the course of a TBRF infection (persistently infected brain tissues as source for spirochetes) when a state of immunosuppression was initiated [50].…”

Section: Discussionmentioning

confidence: 99%

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Rapid clearance of Borrelia burgdorferi from the blood circulation

Liang¹,

Wang

Schorter³

et al. 2020

Parasites Vectors

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Background: Borrelia burgdorferi is a tick-borne spirochete that causes Lyme borreliosis (LB). After an initial tick bite, it spreads from the deposition site in the dermis to distant tissues of the host. It is generally believed that this spirochete disseminates via the hematogenous route. Borrelia persica causes relapsing fever and is able to replicate in the blood stream. Currently the exact dissemination pathway of LB pathogens in the host is not known and controversially discussed. Methods: In this study, we established a strict intravenous infection murine model using host-adapted spirochetes. Survival capacity and infectivity of host-adapted B. burgdorferi sensu stricto (Bbss) were compared to those of B. persica (Bp) after either intradermal (ID) injection into the dorsal skin of immunocompetent mice or strict intravenous (IV) inoculation via the jugular vein. By in vitro culture and PCR, viable spirochetes and their DNA load in peripheral blood were periodically monitored during a 49/50-day course post-injection, as well as in various tissue samples collected at day 49/50. Specific antibodies in individual plasma/serum samples were detected with serological methods. Results: Regardless of ID or IV injection, DNA of Bp was present in blood samples up to day 24 post-challenge, while no Bbss was detectable in the blood circulation during the complete observation period. In contrast to the brain tropism of Bp, Bbss spirochetes were found in ear, skin, joint, bladder, and heart tissue samples of only ID-inoculated mice. All tested tissues collected from IV-challenged mice were negative for traces of Bbss. ELISA testing of serum samples showed that Bp induced gradually increasing antibody levels after ID or IV inoculation, while Bbss did so only after ID injection but not after IV inoculation. Conclusions: This study allows us to draw the following conclusions: (i) Bp survives in the blood and disseminates to the host's brain via the hematogenous route; and (ii) Bbss, in contrast, is cleared rapidly from the blood stream and is a tissue-bound spirochete.

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Section: Discussionmentioning

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Rapid clearance of Borrelia burgdorferi from the blood circulation

Liang¹,

Wang

Schorter³

et al. 2020

Parasites Vectors

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“…In this study, murine skin biopsies were chosen for several reasons. Skin is the primary site to B. burgdorferi infection after a tick bite, and it is well known that B. burgdorferi can establish persistent infection in mice skin [57]. Furthermore, murine skin biopsies have a great advantage in that they are easily available from surplus leftover material from various rodent research studies.…”

Section: Discussionmentioning

confidence: 99%

Ex Vivo Murine Skin Model for B. burgdorferi Biofilm

et al. 2020

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Borrelia burgdorferi, the causative agent of Lyme disease, has been recently shown to form biofilm structures in vitro and in vivo. Biofilms are tightly clustered microbes characterized as resistant aggregations that allow bacteria to withstand harsh environmental conditions, including the administration of antibiotics. Novel antibiotic combinations have recently been identified for B. burgdorferi in vitro, however, due to prohibiting costs, those agents have not been tested in an environment that can mimic the host tissue. Therefore, researchers cannot evaluate their true effectiveness against B. burgdorferi, especially its biofilm form. A skin ex vivo model system could be ideal for these types of experiments due to its cost effectiveness, reproducibility, and ability to investigate host–microbial interactions. Therefore, the main goal of this study was the establishment of a novel ex vivo murine skin biopsy model for B. burgdorferi biofilm research. Murine skin biopsies were inoculated with B. burgdorferi at various concentrations and cultured in different culture media. Two weeks post-infection, murine skin biopsies were analyzed utilizing immunohistochemical (IHC), reverse transcription PCR (RT-PCR), and various microscopy methods to determine B. burgdorferi presence and forms adopted as well as whether it remained live in the skin tissue explants. Our results showed that murine skin biopsies inoculated with 1 × 107 cells of B. burgdorferi and cultured in BSK-H + 6% rabbit serum media for two weeks yielded not just significant amounts of live B. burgdorferi spirochetes but biofilm forms as well. IHC combined with confocal and atomic force microscopy techniques identified specific biofilm markers and spatial distribution of B. burgdorferi aggregates in the infected skin tissues, confirming that they are indeed biofilms. In the future, this ex vivo skin model can be used to study development and antibiotic susceptibility of B. burgdorferi biofilms in efforts to treat Lyme disease effectively.

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“…For example, this relationship has been shown in rodent malaria where the parasite density in the blood is critical for mouse-to-mosquito transmission (59). For B. burgdorferi sl pathogens, the skin rather than the blood is the critical tissue for host-to-tick transmission (60, 61). Field studies on B. afzelii in bank voles and other wild rodents found a positive relationship between the spirochete load in the ear tissues and transmission to larval I. ricinus ticks (62).…”

Section: Discussionmentioning

confidence: 99%

Borrelia afzeliidoes not suppress the development of anti-tick immunity in bank voles

Gomez-Chamorro

Herrera

et al. 2020

Preprint

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Vector-borne pathogens manipulate their vertebrate hosts to enhance their transmission to arthropod vectors. The ability of vertebrate hosts to develop acquired immunity against arthropod vectors represents an existential threat for both the vector and the pathogen. The purpose of the study was to test whether the tick-borne spirochete bacterium Borrelia afzelii could suppress the development of acquired immunity to its tick vector Ixodes ricinus in the bank vole Myodes glareolus, which is an important host for both the tick and the pathogen. We created a group of B. afzelii-infected bank voles and an uninfected control group by exposing lab-reared animals to infected or uninfected ticks. At 1, 2, and 3 months post-infection, all bank voles were infested with larval I. ricinus ticks. The bank voles developed a strong antibody response against tick salivary gland extract proteins. This anti-tick immunity had negative effects on tick fitness traits including engorged larval weight, unfed nymphal weight, larva-to-nymph molting time and larva-to-nymph molting success. Infection with B. afzelii did not suppress the development of acquired immunity against I. ricinus ticks. The development of anti-tick immunity was strongly correlated with a dramatic temporal decline in both the bacterial abundance in the host ear tissues and the host-tick transmission success of B. afzelii. Our study suggests that the development of anti-tick immunity in bank voles has important consequences for the density of infected ticks and the risk of Lyme borreliosis.ImportanceMany pathogens enhance their persistence and transmission by suppressing the immune system of their host. We used an experimental infection approach to test whether the Lyme disease pathogen, Borrelia afzelii, could suppress the development of acquired immunity against its tick vector (Ixodes ricinus) in the bank vole (Myodes glareolus), but found no evidence for this phenomenon. Uninfected and B. afzelii-infected bank voles both developed a strong IgG antibody response against tick salivary gland extract following repeated infestations with I. ricinus ticks. The development of anti-tick immunity was negatively correlated with the abundance of B. afzelii in ear tissue biopsies and with host-to-tick transmission to I. ricinus ticks. Our study suggests that anti-tick immunity in the bank vole reduces the prevalence of this important tick-borne pathogen.

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Identification of Borrelia protein candidates in mouse skin for potential diagnosis of disseminated Lyme borreliosis

Cited by 21 publications

References 66 publications

Rapid clearance of Borrelia burgdorferi from the blood circulation

Rapid clearance of Borrelia burgdorferi from the blood circulation

Ex Vivo Murine Skin Model for B. burgdorferi Biofilm

Borrelia afzeliidoes not suppress the development of anti-tick immunity in bank voles

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