Localization of Borrelia burgdorferi in Murine Lyme Borreliosis by Electron Microscopy

Pachner, Andrew R.; Bašta, Jiří; Delaney, Elizabeth; Hulínská, D

doi:10.4269/ajtmh.1995.52.128

Cited by 48 publications

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“…Studies by various investigators with IS NHPs (11) and rodents by silver staining (7) and electron microscopy (38) have shown that B. burgdorferi has a tropism for the extracellular collagenous matrix of many tissues. This property also has been observed for human Lyme disease, in the skin (8), skeletal muscle (47), and synovium (30).…”

Section: Discussionmentioning

confidence: 99%

Infection and Inflammation in Skeletal Muscle fromNonhuman Primates Infected with Different Genospecies of theLyme Disease SpirocheteBorreliaburgdorferi

et al. 2003

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Lyme borreliosis is a multisystemic disease caused by various genospecies of the spirochete Borrelia burgdorferi. To investigate muscle involvement in the nonhuman primate (NHP) model of Lyme disease, 16 adult Macaca mulatta animals inoculated with strain N40 of B. burgdorferi sensu strictu by syringe or by tick bite or with strain Pbi of B. burgdorferi genospecies garinii by syringe were studied. Animals were necropsied while immunosuppressed on day 50 (two animals each inoculated with B. burgdorferi N40 by syringe and with B. garinii Pbi by syringe) or on day 90, 40 days after immunosuppression had been discontinued (four animals each inoculated with strain N40 by syringe, with strain N40 by tick bite, and with strain Pbi by syringe). Skeletal muscles removed at necropsy were studied by (i) microscopic examination of hematoxylin-eosinstained sections for inflammation and tissue injury; (ii) immunohistochemical and digital image analyses for antibody and complement deposition and cellular inflammation; (iii) Western blot densitometry for the presence of antibodies; and (iv) reverse transcription-PCR for measurement of the spirochetal load or C1q (the first component of the complement cascade) synthesis. The results showed that N40 was more infectious for NHPs than Pbi. NHPs inoculated with N40 but not with Pbi developed myositis. The inflammation in skeletal muscle was more severe in NHPs inoculated with N40 by syringe than in those inoculated by tick bite. The predominant cells in the inflammatory infiltrate were T cells and plasma cells. The deposition of antibody and complement in inflamed muscles from N40-inoculated NHPs was significantly higher than that in Pbiinoculated NHPs. The spirochetal load was very high in the two N40-inoculated NHPs examined while they were immunosuppressed but decreased to minimal levels in the NHPs when immunocompetence was restored. We conclude that myositis can be a prominent feature of Lyme borreliosis depending on the infecting organism and host immune status.

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Section: Discussionmentioning

confidence: 99%

Infection and Inflammation in Skeletal Muscle fromNonhuman Primates Infected with Different Genospecies of theLyme Disease SpirocheteBorreliaburgdorferi

et al. 2003

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“…Os roedores têm sido utilizados como modelos para bioensaios com borrélias (Masuzawa et al 1992, Pachner et al 1995. Foram realizados experimentos com carrapatos avaliando-se potencial de transmissão, patogenicidade de cepas, ultramicroscopia de tecido infectado, desenvolvimento de vacinas, ensaios com drogas e expressão genética de Borrelia sp.…”

Section: V V V V Borreliose Em Animais Sil Borreliose Em Animaisunclassified

“…Foram realizados experimentos com carrapatos avaliando-se potencial de transmissão, patogenicidade de cepas, ultramicroscopia de tecido infectado, desenvolvimento de vacinas, ensaios com drogas e expressão genética de Borrelia sp. (Shih & Spielman 1993, Jobe et al 1994, Pachner et al 1995, Silva & Fikrig 1997.…”

Section: V V V V Borreliose Em Animais Sil Borreliose Em Animaisunclassified

Borrelioses, agentes e vetores

et al. 2000

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1 RESUMO RESUMO RESUMO RESUMO RESUMO.-As borrelioses são enfermidades infecciosas determinadas por espiroquetas do gênero Borrelia, agentes transmissíveis, principalmente, por carrapatos aos animais e/ou ao homem. Nesta revisão são apresentadas e discutidas as enfermidades determinadas por borrélias, bem como as características gerais das espiroquetas, os aspectos relacionados a transmissão por artrópodes, as enfermidades nos animais domésticos e silvestres, quanto aos aspectos bioló-gicos e patológicos, a doença de Lyme como principal zoonose do grupo, a associação de borrélia com outros agentes hematozoários e os métodos diagnósticos e a epidemiologia comparativa entre dados obtidos no Brasil com os de outros países. Estas borrelioses possuem características patológicas, clínicas e epidemiológicas variadas de acordo à região fisiográ-fica, devido à existência de distintas espécies, genoespécies e cepas; estes aspectos variam ainda em função dos artrópodes vetores, da interação vetor-patógeno e dos ecossistemas distintos.TERMOS DE INDEXAÇÃO: Borreliose, carrapatos, Borrelia spp., febre recurrente, borreliose aviária, borreliose bovina, doença de Lyme, aborto epizoótico bovino.

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“…They existed free in the interstitium or were in contact with fibroblasts. The tropism of Borrelia burgdorferi for collagen (3,12) and fibroblasts (11) has been noted in vivo and in vitro. The spirochetes measured were 0.2 urn in diameter.…”

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“…There has been much evidence to explain the mechanism: i.e., B. burgdorferi were found, using electron microscopy, in the vessels of bioptic skin samples from Lyme disease patients (8) and experimentally infected mice (12). Furthermore, B. burgdorferi penetrate into cultured human umbilical vein endothelial cells (2,14).…”

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Ultrastructural Features of Borrelia garinii in Naturally Infected Voles, Clethrionomys rufocanus, with Special Reference to the Relationship with Peripheral Nerves

Yano

Ishiguro²,

Nakata

et al. 2001

Microbiology and Immunology

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Abstract:The ultrastructural features of Borrelia garinii found in the ear tissues of the vole, Clethrionomys rufocanus, are described. The spirochetes were observed in the interstitium of connective tissue or in contactwith fibroblasts and were occasionally situated in the endoneurium of the peripheral nerves. The spirochetes did not injure or enter into the fibroblasts or Schwann cells. The cytotoxicity and migration of the spirochetes are also discussed. Key words: Borrelia garinii, Borrelia infection, Clethrionomys rufocanus, Peripheral nerves, UltrastructureSmall mammals may act as efficient reservoir hosts for Lyme disease spirochetes (16). In a field survey of the prevalence of the disease, ear punches of mammals such as rodents are often used to isolate borreliae (9, 17). We examined natural infection with borreliae among Microtinae voles, Clethrionomys rufocanus, in Hokkaido, Japan (9). The voles were known to be positive for Borrelia garinii, which is widely distributed from Europe to Far East Asia, including Japan (17).Infection by B. garinii in particular is frequently associated with neurological manifestations (1). Since there have been few in vivo studies of B. garinii, information is desirable to understand the pathogenesis of Eurasian Lyme disease. In this study, we detected B. garinii in the ear tissues of naturally infected voles by electron microscopy and found a close relationship between the spirochetes and the peripheral nerves.For isolation and identification of Borrelia, details were reported previously (9). Briefly, 12 C. rufocanus specimens, a common vole species in Hokkaido Prefecture, northern Japan, were captured in October 1994 by using Sherman traps. Five or six pieces of ear punches (about 2 mm in diameter) were removed from each vole, using sterile techniques, and divided into equal portions for culture and electron microscopy. The cul-*Address correspondence to Dr. Yasuhiro Yano, Department of Immunology and Medical Zoology, Fukui Medical University, Matsuoka, Fukui 910-1193, Japan. E-mail: yhyano@fmsrsa. fukui-med.ac.jp 847 turing of the ear punches in Barbour-Stoenner-Kelly II medium showed 5 voles were positive for spirochetes. These isolates (named strain HkCRI-5) were identified as B. garinii based on the monoclonal antibody reactivity and analysis of the restriction fragment length polymorphism pattern. Mixed infections of different borrelial species were not confirmed. Therefore these five voles were thought to be infected only with B. garinii, then selected for the following electron microscopy.The remaining ear punches were fixed in ice-cold 3% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.4) for 6 hr and washed thoroughly in buffer. They were post-fixed with 1% OS04 in buffer for 3.5 hr, dehydrated with acetone, and embedded in epoxy resin. Thin sections (about 80 om) were cut on a microtome and doubly stained with uranyl and lead acetate before observation with an electron microscope (Hitachi H-600, 75 kV).Spirochetes were often observed in the collagenous intercel...

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Localization of Borrelia burgdorferi in Murine Lyme Borreliosis by Electron Microscopy

Cited by 48 publications

References 0 publications

Infection and Inflammation in Skeletal Muscle fromNonhuman Primates Infected with Different Genospecies of theLyme Disease SpirocheteBorreliaburgdorferi

Infection and Inflammation in Skeletal Muscle fromNonhuman Primates Infected with Different Genospecies of theLyme Disease SpirocheteBorreliaburgdorferi

Borrelioses, agentes e vetores

Ultrastructural Features of Borrelia garinii in Naturally Infected Voles, Clethrionomys rufocanus, with Special Reference to the Relationship with Peripheral Nerves

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