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  • 1
    Electronic Resource
    Electronic Resource
    Heterogeneity of Borrelia burgdorferi sensu lato demonstrated by an ospA-type-specific PCR in synovial fluid from patients with Lyme arthritis (1998)
    Springer
    Medical microbiology and immunology 187 (1998), S. 97-102 
    Details
    ISSN: 1432-1831
    Keywords: Key words Lyme borreliosis ; Lyme arthritis ; Polymerase chain reaction ; Outer surface protein A ; Borrelia burgdorferi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Borrelia burgdorferi sensu lato, the etiological agent of Lyme borreliosis, has been divided into three genospecies: B. burgdorferi sensu stricto (OspA-type 1), B. afzelii (OspA-type 2) and B. garinii (OspA-type 3–7). Whereas in Europe B. afzelii (OspA-type 2) is predominant among human skin isolates and B. garinii (OspA-type 3–7) among human CSF isolates, some previous serological studies suggested that Lyme arthritis is also associated with B. burgdorferi sensu stricto in Europe. In the present study we designed ospA type-specific PCRs and identified four different ospA types associated with Lyme arthritis. Our study group consisted of 20 patients with positive serology (ELISA and immunoblotting) and clinical criteria for Lyme arthritis. B. burgdorferi DNA was detected in 13 patients and in none of 10 control patients from synovial fluid. We identified ospA-type 1 (26.6%), ospA-type 2 (33.3%), ospA-type 4 (6.6%) and ospA-type 5 (33.3%). Our conclusion is that in Europe B. burgdorferi sensu lato strains causing Lyme arthritis are considerably heterogeneous and that there is no prevalence of certain genospecies or OspA-types among this strains.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004300050079
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  • 2
    Electronic Resource
    Electronic Resource
    An improved recombinant IgG immunoblot for serodiagnosis of Lyme borreliosis (1999)
    Springer
    Medical microbiology and immunology 188 (1999), S. 139-144 
    Details
    ISSN: 1432-1831
    Keywords: Key wordsBorrelia burgdorferi/afzelii ; Lyme borreliosis ; Recombinant antigen ; Serodiagnosis ; Immunoblot
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We have previously described the use of the following recombinant antigens for serodiagnostic immunoblots: p83/100, p39, OspC and p41 (flagellin) internal fragment [Wilske et al. (1993) Med Microbiol Immunol 182:255–270; Rössler et al. (1997) J Clin Microbiol 35:2752–2758]. In our currently used immunoblot p83/100 is derived from strain PKo (Borrelia afzelii), p39 (BmpA) and OspC from strains PKa2 (B. burgdorferi sensu stricto), PKo and PBi (B. garinii), respectively; the p41 (flagellin) internal fragments were cloned from strains PKo and PBi. In this study we describe the use of two additional recombinantly expressed highly immunogenic proteins Osp17 (derived from PKo) and p58 (derived from PBi). A clinically well-defined panel of sera from 147 Lyme borreliosis patients and 139 controls previously tested by a standardized whole cell lysate immunoblot [Hauser et al. (1997) J Clin Microbiol 35:1433–1444] was investigated in the recombinant immunoblot without (old recombinant immunoblot) and with Osp17 and p58 (new recombinant immunoblot) for IgG antibodies. The sensitivity of the recombinant IgG immunoblot for diagnosis of stage II and stage III could be significantly improved by addition of Osp17 and p58 without loss of specificity. With the exception of sera from patients with erythema migrans the diagnostic sensitivity is comparable to the whole cell lysate IgG immunoblot. The main advantage of the recombinant immunoblot is the easy identification of diagnostic bands, whereas the identification of bands in the whole cell lysate immunoblot is difficult. The recombinant immunoblot is especially suitable where large series of sera need to be investigated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004300050116
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  • 3
    Electronic Resource
    Electronic Resource
    Molecular and immunological characterization of the p83/100 protein of various Borrelia burgdorferi sensu lato strains (1995)
    Springer
    Medical microbiology and immunology 184 (1995), S. 23-32 
    Details
    ISSN: 1432-1831
    Keywords: Lyme borreliosis ; Borrelia burgdorferi ; sensu lato ; p83/100 Antigen ; Molecular analysis ; Phenotypic analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The complete coding regions of the chromosomally encoded p83/100 protein of four Borrelia garinii strains and one Borrelia burgdorferi sensu stricto strain have been amplified by the polymerase chain reaction (PCR), cloned and sequenced. From alignment studies with the deduced amino acid sequences presented here, and five other published p83/100 sequences, the most heterologous region of the p83/100 molecule was identified to be located between amino acid position 390–540. To study the structure of this heterogeneous region, and internal fragment of the p83/100 genes from 11 additional B. burgdorferi sensu lato strains was amplified by PCR. The PCR products were analyzed by DNA sequencing and restriction enzyme analysis. These internal p83/100 fragments varied in size and sequence. Cluster analysis of internal p83/100 fragments, as well as restriction enzyme analysis, revealed three major groups in accordance with grouping into the three species causing Lyme disease. Strains within the same species (six B. burgdorferi sensu stricto and six B. afzelii strains) showed similar p83/100 partial structures. Nevertheless, nine B. garinii strains showed more sequence variations and could be further divided into two major subgroups. One group is represented by OspA serotype 4 strains, the other more heterogeneous group is represented by OspA serotypes 3, 5, 6 and 7 strains. Phenotypic analysis with four p83/100-specific monoclonal antibodies revealed four distinct reactivity patterns. Antibody L100 1B4 recognized a common epitope of B. burgdorferi sensu stricto and B. afzelii. Antibodies L100 17D3 and L100 18B4 were reactive with an epitope shared by strains of all three species. The broadest reactivity was shown by L100 18B4 which, in constrast to L100 17D3, additionally recognized the relapsing fever borreliae B. turicatae and B. hermsii. L100 8B8 detected a subgroup of the B. burgdorferi sensu stricto strains. Since comparison of the p83/100 molecule with sequences from protein databases showed similarities with characteristics of eukaryotic cell structures, the p83/100 might mimic these structures and may, therefore, be involved in the immune escape mechanism of the pathogenic agent of Lyme disease.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00216786
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  • 4
    Electronic Resource
    Electronic Resource
    Osp17, a novel immunodominant outer surface protein of Borrelia afzelii: recombinant expression in Escherichia coli and its use as a diagnostic antigen for serodiagnosis of Lyme borreliosis (1999)
    Springer
    Medical microbiology and immunology 187 (1999), S. 213-219 
    Details
    ISSN: 1432-1831
    Keywords: Key words Outer surface protein ; Osp17 ; Borrelia burgdorferi ; Borrelia afzelii ; Lyme borreliosis ; Serodiagnosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Western blot analyses of the human humoral response of patients with Lyme borreliosis have shown that a 17-kDa protein is an immunodominant protein in late disease. Immune electron microscopy with a monoclonal antibody against this protein revealed that the 17-kDa protein is abundantly expressed on the surface of Borrelia afzelii strain PKo. Therefore, the protein has been renamed outer surface protein (Osp)17. Recombinant Osp17 of strain PKo was expressed in Escherichia coli and purified by chromatography. Immunoblot analysis of human sera showed a comparable sensitivity with recombinant and natural proteins. The DNA sequences of the osp17 genes from different B. afzelii strains were determined. The DNA sequences of the different osp17 homologues (six isolates from skin, three isolates from CSF and one isolate from synovial fluid) had high sequence identities of at least 94%. Using a polyclonal antibody against recombinant Osp17, it was shown that Osp17 expression varied considerably among the investigated B. afzelii strains. As previously also observed for OspA- and OspC-encoding genes, the osp17 gene is present in strains not expressing the respective protein. It has been shown that OspA and OspC expression varies in different environments such as tick and vertebrate host. Studies are underway to examine whether this is also true for Osp17. For diagnostic purposes the use of recombinant Osp17 has the advantage that the amount of Osp17 antigen can be easily standardized for immunoblotting, and that this antigen can be used in a protein-specific enzyme-linked immunosorbent assay.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004300050095
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  • 5
    Electronic Resource
    Electronic Resource
    Active immunization with pC protein ofBorrelia burgdorferi protects gerbils againstB. burgdorferi infection (1992)
    Springer
    Infection 20 (1992), S. 342-349 
    Details
    ISSN: 1439-0973
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Weite Verbreitung und zunehmende Inzidenz der Lyme Borreliose sowie Therapieprobleme bei schweren Erkrankungsformen besonders im Spätstdadium der Infektion sind Gründe für die Entwicklung einer möglichen Alternative zur Antiobitikatherapie wie zum Beispiel Schutzimpfung. Die Schutzwirkung einer aktiven Immunisierung mit rekombinanten OspA und pC gegen die Infektion mitBorrelia burgdorferi wurde bei Gerbils geprüft. Die Tiere wurden mit rekombinanten OspA und pC vomB. burgdorferi-Stamm PKo immunisiert; die Infektion erfolgte vier Wochen nach der Immunisierung mit dem PKo-Stamm (schwache OspA, gute pC Expression). Als Kontrollgruppe dienten infizierte, nicht immunisierte Gerbils. Die immunisierten Tiere bildeten Antikörper gegen rekombinante Vakzine. Die mit pC immunisierten Tiere waren vor der Infektion geschützt, die Kontrollgruppe zeigte eine generalisierte Infektion. Die Immunisierung mit OspA schützte nicht, die Tiere zeigten, wie die Kontrollgruppe Merkmale der Infektion. Die Ergebnisse dieser Studie zeigen, daß pC für eine Immunisierung in Frage kommt. Eine Kombination von OspA und pC scheint für eine effektive Vakzine gegenBorrelia burgdorferi nötig zu sein.
    Notes: Summary Serious infection due toBorrelia burgdorferi and the disseminated infection characteristic of the disease possess unique treatment problems. The wide and still increasing incidence of Lyme borreliosis as well as the problems in treatment call for effective prevention strategies by active immunization. Vaccination experiments were done to determine if active immunization of gerbils with recombinant OspA and pC protects against infection with strains ofB. burgdorferi. Gerbils were vaccinated with recombinant OspA and pC (20 kDa protein) and challenged four weeks later with a clone (derived fromB. burgdorferi strain PKo) which expresses an abundant amount of pC but only little OspA. Non-immunized gerbils challenged with the sameB. burgdorferi strain were used as controls. Both groups of immunized gerbils developed antibodies against the recombinant vaccines. The pC vaccinated group was protected against infection, whereas the OspA vaccinated group showed signs of infection. The non-vaccinated group developed generalised infection. These results show that pC should be considered as a further vaccine candidate and probably needs to be combined with OspA for an efficient vaccine againstB. burgdorferi.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01710681
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