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  • 1
    Electronic Resource
    Electronic Resource
    Molecular analysis and expression of a Borrelia burgdorferi gene encoding a 22kDa protein (pC) in Escherichia coli (1992)
    Oxford, UK : Blackwell Publishing Ltd
    Molecular microbiology 6 (1992), S. 0 
    Details
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: We describe the cloning and expression of the pc gene which encodes a major immunodominant protein of Borrelia burgdorferi, the causative agent of Lyme borreliosis. The pC protein was purified from lysates of B. burgdorferi strain PKo. After tryptic digestion of the pC protein the resulting oligopeptides were applied to a gas-phase sequenator. Thus partial amino acid sequences were obtained. The deduced oligonucleotides were used as hybridization probes. After Southern blotting a reactive band in the 3 kb range of PstI-digested genomic DNA was detected. The insertion of these fragments into pUC vectors finally resulted in pc-positive Escherichia coli clones. The gene (encoding a protein with 212 amino acids) was expressed in E. coli with varying deletions at the 5′ end. A sequence comparison with other outer membrane proteins of B. burgdorferi indicates a processing of pC that is similar to that of lipoproteins.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-2958.1992.tb01495.x
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  • 2
    Electronic Resource
    Electronic Resource
    Borrelia burgdorferi myositis: report of eight patients (1993)
    Springer
    Journal of neurology 240 (1993), S. 278-283 
    Details
    ISSN: 1432-1459
    Keywords: Borrelia burgdorferi ; Lyme disease ; Myositis ; Immunohistology ; Culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Myositis is a rare manifestation of Lyme disease of unknown pathogenesis. This study describes the course of disease in eight patients with Lyme disease, aged 37–70 years, all of whom were suffering from histologically proven myositis. The Clnical, electrophysiological, and myopathological findings are reported. One patient showed signs and symptoms of myositis of all limbs. In six patients myositis was localized in the vicinity of skin lesions, arthritis or neuropathy caused byBorrelia burgdorferi. In another patient suffering from pronounced muscle weakness of the legs and cardiac arrest, inflammation of the myocardium, the conducting system and skeletal muscles was revealed at autopsy. Muscle biopsy revealed lymphoplasmocellular infiltrates combined with few fibre degenerations in three patients. The lymphoplasmocellular infiltrates were found predominantly in the vicinity of small vessels. Several spirochetes were stained in six of seven muscle biopsy samples by means of the immunogold-silver technique. Culturing ofB. Burgdorferi from the muscle biopsy samples was, however, unsuccessful. Antibiotic treatment succeeded in curing the myositis in four of six patients. In one patients signs and symptoms improved. One patient died from cardiac arrest caused by myocarditis and Guillain-Barré syndrome. The outcome is unknown in one patient. Clinical and myopathological findings indicate that Lyme myositis can be caused either by local spreading ofB. burgdorferi or an unknown antigen or toxin from adjacent tissues or haematogenously.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00838161
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  • 3
    Electronic Resource
    Electronic Resource
    Persistent atrioventricular block in Lyme borreliosis (1990)
    Springer
    Journal of molecular medicine 68 (1990), S. 431-435 
    Details
    ISSN: 1432-1440
    Keywords: Lyme borreliosis ; Borrelia burgdorferi spirochete ; Lyme carditis ; Atrioventricular block
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Cardiac manifestations are reported in 0.3%–4.0% of European patients withBorrelia burgdorferi (B.b.) infection. Usually symptoms disappear within 6 weeks. We report a case with persistent impairment of atrioventricular (AV) conduction. Diagnosis was confirmed by demonstration of IgM antibodies and increase of IgG antibody titers against B.b. in serum, by isolation of the spirochete from skin biopsy material and by the typical clinical combination of erythema migrans, Bannwarth syndrome (meningoradiculitis), and complete heart block. Despite immediate antibiotic therapy with ceftriaxone, first degree AV block and second degree block Wenckebach with atrial pacing at 100 beats/minute persisted for 2 years. We conclude, that Lyme carditis can cause long-standing or irreversible AV conduction defects despite adequate and early antimicrobial therapy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01648587
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  • 4
    Electronic Resource
    Electronic Resource
    Recombinant immunoblot in the serodiagnosis of Lyme borreliosis (1993)
    Springer
    Medical microbiology and immunology 182 (1993), S. 255-270 
    Details
    ISSN: 1432-1831
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A recombinant immunoblot was developed for detection of IgM and IgG antibodies in patients with Lyme borreliosis. The recombinant antigens were the chromosomal-encodedBorrelia burgdorferi proteins p100, the flagellin and an internal flagellin fragment thereof as well as the plasmid-encoded outer surface proteins A (OspA) and C (OspC). A panel of 144 sera from patients with Lyme borreliosis (erythema migrans,n = 31; neuroborreliosis state II,n = 60; Lyme arthritis,n = 24 and acrodermatitis chronica atrophicans,n =19) have been investigated and the results have been compared to the immunofluorescence absorption test (IFA-ABS) and to two different enzyme-linked immunosorbent assays [the flagellin ELISA and a newly developed ELISA (OGP-ELISA)]. The two ELISAs were comparable in sensitivity, whereas the IFA-ABS was less sensitive for IgM antibody but equally sensitive for IgG antibody detection. Immunoblot analysis revealed that IgG antibodies are mainly reactive with p 100 and the internal flagellin fragment (sensitivity 51% and 32%, respectively) and rarely with OspC (14%). All patients with late Lyme borreliosis had IgG antibodies against the p100. IgM antibodies were predominantly directed against OspC (43%) and in a lower extent against the internal flagellin fragment and p100 (15% and 13%, respectively). The complete flagellin was not useful due to a high number of unspecific reactions with control sera and the OspA was only exceptionally reactive in Lyme borreliosis patients. The sensitivity of IgM antibody detection could be increased in cases with early Lyme borreliosis from 46% to 65% when the OspC blot was performed in addition to the flagellin ELISA, or from 56% to 65% when performed in addition to the OGP-ELISA. The recombinant blot is, therefore, a valuable diagnostic test to increase sensitivity of early antibody detection and is regarded as a valuable confirmatory test also in late disease.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00579624
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  • 5
    Electronic Resource
    Electronic Resource
    Genetic heterogenity of the genes coding for the outer surface protein C (OspC) and the flagellin of Borrelia burgdorferi (1993)
    Springer
    Medical microbiology and immunology 182 (1993), S. 37-50 
    Details
    ISSN: 1432-1831
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The ospC gene coding for the outer surface protein OspC and the fla gene coding for the flagellin have been investigated in three different Borrelia burgdorferi sensu lato strains. These strains (the North American strain B31 and the European strains PKo and PBi) derive from various biological sources (lxodes dammini, human skin and human CSF) and belong to three different B. burgdorferi OspA serotypes and genospecies (OspA serotype 1, B. burgdorferi sensu stricto; OspA serotype 2, group VS461 and OspA serotype 4, B. garinii, respectively). The ospC and fla genes of the respective strains have been amplified by polymerase chain reaction, cloned in pUC8 and sequenced. The fla as well as the ospC genes were different among the three strains investigated. In general the fla genes are more conserved than the ospC genes. The fla genes have the same length of 1008 nucleotides coding for proteins of 336 amino acids, whereas the ospC genes differ in length. The ospC genes of strains B31, PKo and PBi have 630, 636 and 621 nucleotides encoding proteins of 210, 212 and 207 amino acids, respecctively. The ospC genes exhibit sequence identities between 70% and 74% among each other, sequence identities of the fla genes are in the range 96–97%. The ospC genes could be expressed in Escherichia coli to obtain proteins with and without leader peptides. The expression of the fla gene and an internal gene fragment resulted in the complete flagellin protein and a truncated protein (amino acids 129–251). The different ospC and fla gene products were immunoreactive with monoclonal antibodies and human sera and, thus, enlarge the spectrum of recombinant antigens to improve antibody detection in patients with Lyme borreliosis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00195949
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  • 6
    Electronic Resource
    Electronic Resource
    Genetic polymorphism of the gene encoding the outer surface protein A (OspA) of Borrelia burgdorferi (1992)
    Springer
    Medical microbiology and immunology 181 (1992), S. 57-70 
    Details
    ISSN: 1432-1831
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The genes coding for the outer surface protein (OspA) of Borrelia burgdorferi, the causative agent of Lyme borreliosis have been cloned and sequenced. Two German strains (skin isolate PKo and cerebrospinal fluid isolate PBi) have been analyzed. Using an OspA-specific monoclonal antibody (L32 2E7) for immunological screening of a genomic pUC 18 library of B. burgdorferi strain PKo, an OspA-producing clone was detected and subclones containing the open reading frame were constructed. The gene coding for the OspA protein of B. burgdorferi strain PBi was amplified using polymerase chain reaction (PCR) and cloned in pUC8. The open reading frame of both ospA genes consists of 822 nucleotides corresponding to a protein of 273 amino acids. Both proteins have a calculated molecular mass of 29.6 kDa. Molecular analysis revealed significant differences between each other and to already-published sequences of ospA of B. burgdorferi strains B31, ZS7 and N40 (the ospA genes of B31, ZS7 and N40 are nearly identical). The deduced amino acid sequences of the OspA protein of strains PKo and PBi showed a homology of 83% to each other and 77% and 80%, respectively, to OspA protein of strain B 31. The three proteins contain a variable middle region, whereas the N and the C terminus are conserved. This unexpected high dissimilarity of the ospA genes may be important in respect to vaccination studies and diagnostic procedures (i.e., development of PCR primers or serodiagnostic antigens). Moreover, the molecular heterogeneity of OspA confirms three out of seven immunologically defined OspA serotypes of a recently proposed OspA serotyping system.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00189424
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  • 7
    Electronic Resource
    Electronic Resource
    Immunoblot using recombinant antigens derived from different genospecies of Borrelia burgdorferi sensu lato (1994)
    Springer
    Medical microbiology and immunology 183 (1994), S. 43-59 
    Details
    ISSN: 1432-1831
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Immunodominant proteins are variable in molecular and antigenic structure among different genospecies of Borrelia burgdorferi sensu lato. We have recently developed an immunoblot using five recombinant antigens: the chromosomal-encoded B. burgdorferi proteins p 100, the flagellin and an internal flagellin fragment thereof, and the plasmid-encoded outersurface proteins A (OspA) and C (OspC). In the present study the same antigens (derived from strain PKo, genospecies B. afzelii) were compared with the homologous recombinant proteins from strain B31 (genospecies B. burgdorferi sensu stricto) and with OspA, OspC and the internal flagellin fragment from strain PBi (genospecies B. garinii). Patients with neuroborreliosis (n=28) and patients with acrodermatitits chronica atrophicans (n=20) were investigated in the IgG immunoblot; the IgM immunoblot was performed only in patients with neuroborreliosis. There was a small increase in the detection rate of OspA-specific IgG or IgM antibodies using the different variants of recombinant OspA; however, OspA remained an insensitive antigen for antibody detection in Lyme borreliosis. The same was true to OspC-specific IgG antibodies. The sensitivity of OspC, which is the immunodominant antigen for IgM antibody detection, could not be increased using recombinant antigens derived from different strains. However, some sera which were negative in the recombinant immunoblot reacted with OspC in the conventional immunoblot using B. burgdorferi whole cell lysate as antigen. The most unexpected finding was the high degree of immunological heterogeneity of the internal flagellin fragments: IgG antibodies were detected in 18 of 48 patients using B31 fragments, in 25 of 48 using PKo fragments, in 23 of 48 using PBi fragments versus 33 of 48 when the three recombinant proteins were combined. PKo-derived fragments were more sensitive for antibody detection in patients with acrodermatitis chronica atrophicans, B31- and PBi-derived fragments for antibody detection in patients with neuroborreliosis. This is in agreement with the fact that isolates from patients with neuroborreliosis are predominantly belonging to the genospecies B. burgdorferi sensu stricto and B. garinii. For detection of IgM antibodies in sera from patients with neuroborreliosis, recombinant internal fragments derived from strains B31 and PBi were more sensitive than the PKo-derived fragment. The best discrimination between neuroborreliosis sera and control sera was achieved when the IgM blot was performed using recombinant internal flagellin fragments derived from strains PKo and PBi and OspC derived from B31 or PKo.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00193630
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  • 8
    Electronic Resource
    Electronic Resource
    Persistence of borrelia burgdorferi and histopathological alterations in experimentally infected animals. A comparison with histopathological findings in human Lyme disease (1990)
    Springer
    Infection 18 (1990), S. 332-341 
    Details
    ISSN: 1439-0973
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Das Humanisolat P/Ko derBorrelia burgdorferi ist infektiös für Gerbils, wir konnten damit eine 100%ige Infektion der Tiere erreichen. Die Isolierung derB. burgdorferi aus verschiedenen Organen, sechs Monate nach der Infektion, bestätigt eine generalisierte Infektion und zeigt, daß Borrelien in diesen Tieren über Monate persistieren können. Die Spirochätemie wurde 14 Tage beobachtet und erfolgte in zwei Intervallen. Die intraperitoneale Inokulation derB. burgdorferi an sechs Gerbils der Gruppen A und B (infiziert mit 25. bzw. 5. Subkultur des Erregers) verursachte signifikante histopathologische Veränderungen in der Mehrzahl der Organsysteme und dem umgebenden Gewebe. Die histologischen Befunde zeigen perivaskuläre, entzündliche Reaktionen mit Lymphozyten, Histiozyten, Plasmazellen und eosinophilen Leukozyten. Bei Kontrolltieren wurden keine Zeichen von Entzündungen in keinem der Organe festgestellt. Die histopathologischen Ergebnisse der Tiergruppen A und B sind nahezu identisch. Die Persistenz derB. burgdorferi und die hohe Zahl der betroffenen Organsysteme in dieser Versuchsserie kann mit Beobachtungen an Patienten mit Lyme-Borreliose verglichen werden. Die Ergebnisse dieser Studie bestätigen, daß Gerbils als Tiermodell für Studien der Pathogenese der Lyme-Borreliose sowie andereIn-vivo-Studien sehr geeignet sind.
    Notes: Summary Gerbils appear to be susceptible to infection by human isolates ofBorrelia burgdorferi; we obtained 100% infection. Isolation of theB. burgdorferi from different organs six months post infection causes a generalized infection thus demonstrating that borreliae persist in these animals for a long period. Spirochetemia was present for 14 days, apparently in two intervals. TheBorrelia burgdorferi specific antibody titers increased with time after infection thus indicating the persistence of spirochetes. The intraperitoneal inoculation of theB. burgdorferi to six gerbils of groups A and B induced significant histopathologic changes in most of the major organ systems and their surrounding adipose and fibrous connective tissues. The infiltrates consisted mainly of lymphocytes and histiocytes. Various numbers of plasma cells, eosinophils and high numbers of mast cells were also present. Three further animals which served as controls displayed no histological signs of inflammation in any organ system. No significant differences were noted between the histopathological findings seen in the animals of groups A and B (infected with cells from subcultures no. 25 and with no. 5, respectively). The persistence ofB. burgdorferi and the high number of organs involved with slight to severe signs of inflammation in this series can be compared to persistence and to the multiorgan involvement seen in human Lyme disease. Thus gerbils can serve as suitable experimental animals to study the pathogenesis of Lyme disease and the extent of organ damage caused byB. burgdorferi.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01646399
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  • 9
    Electronic Resource
    Electronic Resource
    Immunodominant proteins of Borrelia burgdorferi, the etiological agent of lyme borreliosis (1991)
    Springer
    World journal of microbiology and biotechnology 7 (1991), S. 130-136 
    Details
    ISSN: 1573-0972
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Lyme borreliosis, a multisystem disorder involving the skin, the nervous system, the heart, the joints and many other organs, is a worldwide infectious disease which is transmitted by ticks of the Ixodes complex. Most frequently diagnosis is accomplished by detection of antibodies because the Borrelia are difficult to cultivate. Present serodiagnostic methods, however, are impaired by low sensitivity and unspecific reactions. The selection of immunodominant antigens with low cross-reactivity to other bacteria should improve antibody detection. Borrelia burgdorferi proteins have been analysed for cross-reactivity with immune sera from unrelated bacteria, and sera from patients with different stages of the disease. Suitable antigens for improving serodiagnosis have been detected and are reported here. In view of the immunological heterogeneity of Borrelia proteins, sensitivity of antibody detection may possibly be increased by using recombinant antigens derived from different strains. Immunization with recombinant OspA (a flagellum-associated protein) from a North American isolate protected mice from the challenge with three North American isolates. However, for development of an effective vaccine (especially in Europe), the heterogeneity of OspA has to be considered.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00328982
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  • 10
    Electronic Resource
    Electronic Resource
    Culture ofBorrelia burgdorferi on six solid media (1991)
    Springer
    European journal of clinical microbiology & infectious diseases 10 (1991), S. 1076-1079 
    Details
    ISSN: 1435-4373
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Six solid substrates were compared for their support of growth of ten isolates ofBorrelia burgdorferi, the causative agent of Lyme borreliosis. The following substrates with or without rabbit serum and bovine serum albumin were tested: BSK agar, BHIAM agar, TAROM agar, MEM agar, MKP agar and PMR agar. After incubation in a candle jar and a GasPak for two to four weeks,Borrelia colonies were counted and characterized. Colony morphology was related more to the growth substrate than to the characteristics of the variousBorrelia burgdorferi isolates. Culture on PMR agar resulted in the highest recovery rate and the best colony formation, with a size variation of 0.3–3.0 mm. Culture ofBorrelia burgdorferi on solid media may facilitate the diagnosis of Lyme borreliosis by enabling easier biological and genetical analysis of isolates and more accurate differentiation of strains.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01984935
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