Experimental Lyme Arthritis in Rats Infected with Borrelia burgdorferi

Barthold, Stephen W.; Moody, Kathleen D.; Terwilliger, Gordon A.; Duray, Paul H.; Jacoby, Robert O.; Steere, Allen C.

doi:10.1093/infdis/157.4.842

Cited by 123 publications

(68 citation statements)

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“…In all but one case, strain N40-inoculated TISP NHPs had Ͻ1 spirochete/g of RNA. No spirochetes were detected in any of the skeletal muscles examined from IC NHPs (four of four samples had Ct values of 40) 5 ), respectively. Inflammation in skeletal muscle.…”

Section: Methodsmentioning

confidence: 93%

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: Methodsmentioning

confidence: 93%

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

et al. 2003

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“…This strain was chosen because of its plasmid content, and the disruption of the chbC locus with the coumermycin resistance marker is not predicted to affect transposition. A3 and N40 are infectious strains; A3 is a clonal derivative of the type strain, B31, and contains all plasmids except cp9 (11), and N40 was isolated from a tick obtained in Westchester County, N.Y. (2,3). The plasmid content of N40 has not been determined.…”

Section: Methodsmentioning

confidence: 99%

Genome-Wide Transposon Mutagenesis of Borrelia burgdorferi for Identification of Phenotypic Mutants

Stewart¹,

Hoff²,

Fischer

et al. 2004

Appl Environ Microbiol

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The spirochete Borrelia burgdorferi is the causative agent of Lyme disease, the leading vector-borne illness in the United States. Many of the genetic factors affecting spirochete morphology and physiology are unknown due to the limited genetic tools available and the large number of open reading frames with unknown functions. By adapting a mariner transposon to function in B. burgdorferi, we have developed a random mutagenesis system that tags the mutated locus for rapid identification. Transposition occurs at saturating levels in B. burgdorferi and appears to be random, targeting both linear and circular replicons. By combining the transposon system with a screen for factors affecting growth rate, mutations were readily identified in genes putatively involved in cell division and chemotaxis and a hypothetical open reading frame involved in outer membrane integrity. The successful adaptation of a mariner transposon to function in B. burgdorferi should aid in identifying virulence factors and novel gene products related to spirochete physiology.Lyme disease is the leading vector-borne illness in the United States and is caused by the spirochete Borrelia burgdorferi. The class Spirochaetes contains many significant pathogens of humans and other animals, including Treponema pallidum (agent of syphilis), Treponema denticola (associated with periodontal disease), Brachyspira hyodysenteriae (associated with swine dysentery), Brachyspira pilosicoli (associated with porcine intestinal spirochetosis), Leptospira interrogans (associated with leptospirosis), and Borrelia spp. (associated with relapsing fever) (27). However, many of the genetic tools available for members of the Enterobacteriaceae have not yet been developed for the spirochetes, limiting the experimental identification of virulence factors and genes specific to spirochete physiology. Identifying new virulence factors and functions of hypothetical open reading frames (ORFs) in B. burgdorferi has been further hindered by the low efficiency of targeted allelic exchange (for a review, see reference 25).An efficient and random mutagenesis system would facilitate functional identification of the large number of unknown ORFs identified in the genome sequence of B. burgdorferi (8,12). Some transposon systems are capable of achieving saturation mutagenesis and are nearly random in their insertion sites, and the mutated locus is marked by the transposon insertion and therefore easily identified. Specifically, transposons of the mariner family have been used successfully for mutagenesis of a diverse range of organisms, including eukaryotes, archaea, and both gram-positive and gram-negative bacteria (1,14,16,21,29). The mariner elements do not require host cofactors for transposition, likely contributing to their wide host range (15). Further, mariner transposition is virtually random, requiring only a TA dinucleotide for target specificity. In addition, Lampe and colleagues derived hyperactive transposase mutants of Himar1 (a member of the mariner family), increasing...

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“…B. burgdorferi B31 clones A1.4 and A3H.3 were derived from individual colonies of isolate B31 plated on solid Barbour-Stoener-Kelly medium (37) and cultured for no more than three passages. B. burgdorferi N40 (obtained from S. Barthold, Yale University, New Haven, Conn.) is a wild-type isolate, originally obtained from a tick in Westchester, N.Y. (10), that has been cloned by limiting dilution (9) and has been passaged in culture fewer than five times. B. burgdorferi cells were grown at 34ЊC in BSK-H medium (Sigma, St. Louis, Mo.)…”

Section: Methodsmentioning

confidence: 99%

A family of genes located on four separate 32-kilobase circular plasmids in Borrelia burgdorferi B31

1996

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We have identified four loci in Borrelia burgdorferi B31 that contain open reading frames capable of encoding six proteins that are related to the antigenic proteins OspE and OspF. We have designated these proteins Erp, for OspEF-related protein, and named their respective genes erp. The erpA and erpB genes are linked, as are erpC and erpD, and the pairs probably constitute two operons. The erpG and erpH genes appear to be monocistronic. The ErpA and ErpC proteins are expressed by B. burgdorferi B31 in culture and are recognized by a polyclonal antiserum raised against the OspE protein of B. burgdorferi N40. The four erp loci are each located on different 32-kb circular plasmids that contain additional DNA sequences that are homologous to each other and to an 8.3-kb circular plasmid of B. burgdorferi sensu lato Ip21. All four 32-kb plasmids can be maintained within a single bacterium, which may provide a model for the study of plasmid replication and segregation in B. burgdorferi.Borrelia burgdorferi is a member of the order Spirochaetales, the spirochetes, an order that is phylogenetically and morphologically distinct from such well-characterized bacteria as Escherichia coli and Bacillus subtilis (8,24,34,35,53). B. burgdorferi bacteria naturally contain a wide variety of plasmids (4,13,25,40,42,45,47,54), yet nothing is known about the mechanisms by which plasmids are maintained in these bacteria. Several observations suggest that the small DNA species of B. burgdorferi and other spirochetes may be fundamentally different from the plasmids of other bacteria, the most significant being that there are linear plasmids in B. burgdorferi (4,5,7,13,22,23,42). Also, two B. burgdorferi genes involved in purine biosynthesis are located on a circular plasmid (29), which appears to be inconsistent with the classical definition of a plasmid as a nonessential extrachromosomal element. Other plasmids of B. burgdorferi appear to be capable of undergoing recombination to form multimeric plasmids (25, 31), and there have been reports of DNA sequences located on several different plasmids within the same bacterium (46,47,(54)(55)(56).B. burgdorferi is the causative agent of Lyme disease, an increasingly common ailment of humans and several other mammals (15,48). Infection of a mammal by B. burgdorferi is generally accompanied by the production of antibodies directed against a limited number of bacterial antigens (17,19,41,52), several of which have been identified as surface-exposed lipoproteins. At least three of these lipoproteins, OspC, OspE, and OspF, are differentially expressed by B. burgdorferi, being produced in greater quantities by cultured bacteria that are shifted from 23 to 32ЊC than by those maintained at the lower temperature (43, 49). For OspC, at least, this appears to be related to induction of specific protein synthesis by the spirochete in ticks following feeding upon a warm-blooded animal (43). Temperature-dependent differential expression of bacterial proteins that are involved in host infection has been o...

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Experimental Lyme Arthritis in Rats Infected with Borrelia burgdorferi

Cited by 123 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

Genome-Wide Transposon Mutagenesis of Borrelia burgdorferi for Identification of Phenotypic Mutants

A family of genes located on four separate 32-kilobase circular plasmids in Borrelia burgdorferi B31

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