1 Journal of Neuroinflammation 2008 Vol: 5(1):40. DOI: 10.1186/1742-2094-5-40

Persisting atypical and cystic forms of Borrelia burgdorferi and local inflammation in Lyme neuroborreliosis

The long latent stage seen in syphilis, followed by chronic central nervous system infection and inflammation, can be explained by the persistence of atypical cystic and granular forms of Treponema pallidum. We investigated whether a similar situation may occur in Lyme neuroborreliosis. Atypical forms of Borrelia burgdorferi spirochetes were induced exposing cultures of Borrelia burgdorferi (strains B31 and ADB1) to such unfavorable conditions as osmotic and heat shock, and exposure to the binding agents Thioflavin S and Congo red. We also analyzed whether these forms may be induced in vitro, following infection of primary chicken and rat neurons, as well as rat and human astrocytes. We further analyzed whether atypical forms similar to those induced in vitro may also occur in vivo, in brains of three patients with Lyme neuroborreliosis. We used immunohistochemical methods to detect evidence of neuroinflammation in the form of reactive microglia and astrocytes. Under these conditions we observed atypical cystic, rolled and granular forms of these spirochetes. We characterized these abnormal forms by histochemical, immunohistochemical, dark field and atomic force microscopy (AFM) methods. The atypical and cystic forms found in the brains of three patients with neuropathologically confirmed Lyme neuroborreliosis were identical to those induced in vitro. We also observed nuclear fragmentation of the infected astrocytes using the TUNEL method. Abundant HLA-DR positive microglia and GFAP positive reactive astrocytes were present in the cerebral cortex. The results indicate that atypical extra- and intracellular pleomorphic and cystic forms of Borrelia burgdorferi and local neuroinflammation occur in the brain in chronic Lyme neuroborreliosis. The persistence of these more resistant spirochete forms, and their intracellular location in neurons and glial cells, may explain the long latent stage and persistence of Borrelia infection. The results also suggest that Borrelia burgdorferi may induce cellular dysfunction and apoptosis. The detection and recognition of atypical, cystic and granular forms in infected tissues is essential for the diagnosis and the treatment as they can occur in the absence of the typical spiral Borrelia form.

Mentions
Figures
Figure 1: Characteristic morphology of Borrelia burgdorferi seen by various techniques following one week of culture in BSKII medium Figure 2: Atypical forms of Borrelia burgdorferi (B31 strain) spirochetes induced by harmful culture conditions Figure 3: Rolled and cystic forms of Borrelia burgdoferi spirochetes observed after one week of culture in medium to which Thioflavin S had been added Figure 4: Atypical and cystic Borrelia forms following 1 week exposure of primary neuronal and astrocytic cultures to Borrelia burgdorferi Figure 5: Atypical cystic spirochetes in the medium of neuronal and astrocytic cultures following 1 week exposure to Borrelia burgdorferi Figure 6: Recovery of the typical vegetative form of spirochetes re-cultured in BSK II medium and nuclear fragmentation of rat primary astrocytes exposed to Borrelia burgdorferi Figure 7: Extra- and intracellular atypical and cystic forms of spirochetes in the cerebral cortex of a patient with pathologically and serologically confirmed chronic Lyme neuroborreliosis where Borrelia burgdorferi sensu stricto was cultivated from the brain Figure 8: Chronic neuroinflammation in the frontal cortex of a patient with Lyme neuroborreliosis
Altmetric
References
  1. F Schaudinn; S Hoffman Über Spirochaeta pallida bei Syphilis und die Unterschiede dieser Form gegenuber anderen Arten dieser Gattung Berlin Klin Wochschr 42, 673-675 (1905) .
    • . . . The similarity of clinical and pathological manifestations of syphilis caused by Treponema pallidum 1 and Lyme disease caused by Borrelia burgdorferi 2 is well established . . .
    • . . . As early as 1905 it was suspected that the classical spiral (vegetative) form was not the only one that spirochetes could assume 14 . . .
  2. W Burgdorfer; AG Barbour; SF Hayes; JL Benach; E Grunwaldt; JP Davis Lyme disease-a tick-borne spirochetosis? Science 216, 1317-1319 (1982) .
    • . . . The similarity of clinical and pathological manifestations of syphilis caused by Treponema pallidum 1 and Lyme disease caused by Borrelia burgdorferi 2 is well established . . .
  3. J Miklossy; K Khalili; L Gern; RL Ericson; P Darekar; L Bolle; J Hurlimann; BJ Paster Borrelia burgdorferi persists in the brain in chronic Lyme neuroborreliosis and may be associated with Alzheimer disease J Alzheimers Dis 6, 639-649 (2004) .
    • . . . In analogy to Treponema pallidum, Borrelia burgdorferi persists in the brain in chronic Lyme neuroborreliosis 3 . . .
    • . . . Intracellular localization of Borrelia burgdorferi was observed in macrophages and keratinocytes in the skin 14 and in neurons and glial cells in vitro and in vivo 3495051. . . .
    • . . . Borrelia burgdorferi spirochetes strains B31 and ADB1 3515253 were cultivated in BSK II medium 54 . . .
    • . . . To infect neurons and astrocytes, Borrelia spirochetes of the virulent strains B31 and ADB1, the latter having been cultured from the brain of a patient with concurrent Lyme neuroborreliosis and AD 3, were employed . . .
    • . . . Brains of three patients with pathologically and serologically confirmed Lyme neuroborreliosis and concurrent AD were analyzed 3 . . .
    • . . . The following anti-Borrelia burgdorferi antibodies were used: monoclonal anti-OspA (H5332, H3T5, Symbicom, 1:50) and anti-flagellin (G 9724, H605, Symbicom, 1:50), polyclonal B65302R (Biodesign, 1:100) and BB-1017 (1:500) 3 antibodies . . .
    • . . . The specificity of these mono- and polyclonal antibodies was previously tested by Western blot analysis 3. . . .
    • . . . Panels C and E were reprinted from panels F and D of Figure 5 of Mikossy et al., 2004 3, with permission from IOS Press. . . .
    • . . . Recently we reported evidence that Borrelia burgdorferi can also persist in the brain in chronic Lyme neuroborreliosis and, in analogy to Treponema pallidum, may cause dementia, cortical atrophy and amyloid deposition 34951 . . .
    • . . . Like for Treponema pallidum in neurosyphilis, atypical and cystic forms of Borrelia burgdorferi were also observed intracellularly in the brains of these patients, as it has previously been documented 351 . . .
    • . . . That Borrelia burgdorferi was successfully cultivated from brains of the three patients with Lyme neuroborreliosis in BSK-II medium where pleomorphic and cystic forms were observed in the brain 351527273 suggests that at least part of the persisting spirochetes are viable . . .
    • . . . The clinical and the pathological hallmarks of Alzheimer's disease, including beta-amyloid deposition are also present in the atrophic form of general paresis and in tertiary Lyme neuroborreliosis 363727374 . . .
  4. K Herxheimer Zur Kenntnis der Spirochaeta Pallida München med Wochschr 53, 310-312 (1905) .
    • . . . As early as 1905 it was suspected that the classical spiral (vegetative) form was not the only one that spirochetes could assume 14 . . .
  5. L Jacquet; A Sézary Des formes atypiques et dégénératives du tréponéme pâle Bull mem Soc Med Hop Par 24, 114 (1907) .
    • . . . Transformation of various types of spirochetes into cystic forms through end knob, loop, ring-shaped and spherule formation has since been repeatedly reported 5678910 . . .
  6. AS Warthin; RE Olson The granular transformation of Spirochaeta pallida in aortic focal lesions Am J Syphilis 14, 433-437 (1930) .
    • . . . Transformation of various types of spirochetes into cystic forms through end knob, loop, ring-shaped and spherule formation has since been repeatedly reported 5678910 . . .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
    • . . . The occurrence of pleomorphic forms of Treponema pallidum in the brain in general paresis and their abundance in juvenile paresis is well documented 618264142. . . .
  7. O Brorson; SH Brorson Transformation of cystic forms of Borrelia burgdorferi to normal mobile spirochetes Infection 25, 240-246 (1997) .
    • . . . Transformation of various types of spirochetes into cystic forms through end knob, loop, ring-shaped and spherule formation has since been repeatedly reported 5678910 . . .
  8. O Brorson; S Brorson In vitro conversion of Borrelia burgdorferi to cystic forms in spinal fluid, and transformation to mobile spirochetes by incubation in BSK-H medium Infection 26, 144-150 (1998) .
    • . . . Transformation of various types of spirochetes into cystic forms through end knob, loop, ring-shaped and spherule formation has since been repeatedly reported 5678910 . . .
    • . . . Reconversion of cystic Borrelia burgdorferi into the typical spiral form has been demonstrated in vitro and in vivo 8103140. . . .
    • . . . These results are in harmony with previous observations 85566 . . .
    • . . . This has also been observed in various types of spirochetes [e.g. 113870] including Borrelia burgdorferi 8 by transmission electron microscopy analyses . . .
    • . . . The size of such cysts depends on the number of spirochetes packed inside of the cyst 8 . . .
  9. R Murgia; C Piazzetta; M Cinco Cystic forms of Borrelia burgdorferi sensu lato: induction, development, and the role of RpoS Wien Klin Wochenschr 114, 574-579 (2002) .
    • . . . Transformation of various types of spirochetes into cystic forms through end knob, loop, ring-shaped and spherule formation has since been repeatedly reported 5678910 . . .
  10. I Gruntar; T Malovrh; R Murgia; M Cinco Conversion of Borrelia garinii cystic forms to motile spirochetes in vivo APMIS 109, 383-838 (2001) .
    • . . . Transformation of various types of spirochetes into cystic forms through end knob, loop, ring-shaped and spherule formation has since been repeatedly reported 5678910 . . .
    • . . . Reconversion of cystic Borrelia burgdorferi into the typical spiral form has been demonstrated in vitro and in vivo 8103140. . . .
  11. T Umemoto; I Namikawa; Z Yoshii; H Konishi An internal view of the spherical body of Treponema macrodentium as revealed by scanning electron microscopy Microbiol Immunol 26, 191-198 (1982) .
    • . . . Agglomeration of spirochetes into colonies 11121314, enclosing numerous cystic forms, has been observed both in vitro and in vivo 12. . . .
    • . . . This has also been observed in various types of spirochetes [e.g. 113870] including Borrelia burgdorferi 8 by transmission electron microscopy analyses . . .
  12. T Umemoto; I Namikawa; M Yamamoto Colonial morphology of treponemes observed by electron microscopy Microbiol Immunol 28, 11-22 (1984) .
    • . . . Agglomeration of spirochetes into colonies 11121314, enclosing numerous cystic forms, has been observed both in vitro and in vivo 12. . . .
  13. TJ Kurtti; UG Munderloh; RC Johnson; GG Ahlstrand Colony formation and morphology in Borrelia burgdorferi J Clin Microbiol 25, 2054-2058 (1987) .
    • . . . Agglomeration of spirochetes into colonies 11121314, enclosing numerous cystic forms, has been observed both in vitro and in vivo 12. . . .
  14. E Aberer; A Kersten; H Klade; C Poitschek; W Jurecka Heterogeneity of Borrelia burgdorferi in the skin American Journal of Dermatopathology 18, 571-579 (1996) .
    • . . . Agglomeration of spirochetes into colonies 11121314, enclosing numerous cystic forms, has been observed both in vitro and in vivo 12. . . .
    • . . . Their occurrence has been reported in skin lesions 14, in an ex vivo system in tonsil tissue 47 and on silver stained hippocampus section in a patient with concurrent Alzheimer disease (AD) and Lyme neuroborreliosis 48 . . .
    • . . . Intracellular localization of Borrelia burgdorferi was observed in macrophages and keratinocytes in the skin 14 and in neurons and glial cells in vitro and in vivo 3495051. . . .
  15. CF Garon; DW Dorward; MD Corwin Structural features of Borrelia burgdorferi – the Lyme disease spirochete: silver staining for nucleic acids Scanning Microsc Suppl 3, 109-115 (1989) .
    • . . . In Borrelia burgdorferi these free vesicular or granular structures contain spirochetal surface proteins and linear and circular DNA 1516. . . .
  16. S Bergstrom; CF Garon; AG Barbour; J MacDougall Extrachromosomal elements of spirochetes Res Microbiol 143, 623-628 (1992) .
    • . . . In Borrelia burgdorferi these free vesicular or granular structures contain spirochetal surface proteins and linear and circular DNA 1516. . . .
  17. A Kersten; C Poitschek; S Rauch; E Aberer Effects of penicillin, ceftriaxone, and doxycycline on the morphology of Borrelia burgdorferi Antimicrob Agents Chemother 39, 1127-1133 (1995) .
    • . . . Granular disintegration of spirochetes resulting in a chain of fine granules also occurs under adverse conditions 171819202122 . . .
  18. HB Fantham; MA Cantab Spirochaetes and their granule phase Brit Med J 1, 409-411 (1916) .
    • . . . Granular disintegration of spirochetes resulting in a chain of fine granules also occurs under adverse conditions 171819202122 . . .
    • . . . The occurrence of pleomorphic forms of Treponema pallidum in the brain in general paresis and their abundance in juvenile paresis is well documented 618264142. . . .
  19. E Hindle The life cycle of the organism of syphilis Lancet 2, 1011 (1912) .
    • . . . Granular disintegration of spirochetes resulting in a chain of fine granules also occurs under adverse conditions 171819202122 . . .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
  20. J Ewing Note on involution forms of Spirochaete pallida in gummata Proc N Y Pathol Soc 7, 166-171 (1908) .
    • . . . Granular disintegration of spirochetes resulting in a chain of fine granules also occurs under adverse conditions 171819202122 . . .
  21. A Balfour The infective granule in certain protozoal infections, as illustrated by the spirochaetosis of Sudanese fowl Brit Med J 1, 752 (1911) .
    • . . . Granular disintegration of spirochetes resulting in a chain of fine granules also occurs under adverse conditions 171819202122 . . .
  22. C Nicolle L'evolution des spirochetes et le mecanisme de la crise dans les spirochetoses Arch Inst Pasteur 16, 207-217 (1927) .
    • . . . Granular disintegration of spirochetes resulting in a chain of fine granules also occurs under adverse conditions 171819202122 . . .
  23. WB Leishman An address on the mechanism of infection in tick fever, and on hereditary transmission of Spirochaeta duttoni in the tick Lancet 133, 11-14 (1911) .
    • . . . Minute granules are liberated from the periplasmic sheath through budding and extrusion, which may multiply and may be transmissible 232425262728293031 . . .
  24. WB Leishman A note on the "granule clumps" found in Ornithodorus moubata and their relation to the spirochaetes of African relapsing fever (tick fever) Ann Inst Pasteur 32, 49-59 (1918) .
    • . . . Minute granules are liberated from the periplasmic sheath through budding and extrusion, which may multiply and may be transmissible 232425262728293031 . . .
  25. WB Leishman The Horace Dobell lecture on an experimental investigation of Spirochaeta duttoni, the parasite of tick fever Lancet 2, 1237-1244 (1920) .
  26. Y Manouélian Syphilis héréditaire et formes évolutives du tréponème C rend Acad sc 190, 332 (1930) .
    • . . . Minute granules are liberated from the periplasmic sheath through budding and extrusion, which may multiply and may be transmissible 232425262728293031 . . .
    • . . . Their presence in syphilitic patients was regarded as confirmatory of the syphilitic nature of the lesions even in the absence of classical spiral forms 262730 . . .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
    • . . . The occurrence of pleomorphic forms of Treponema pallidum in the brain in general paresis and their abundance in juvenile paresis is well documented 618264142. . . .
  27. Y Manouélian Gommes syphilitiques et formes anormales du treponemes; Ultra-virus syphilitiques C rend Soc biol 104, 249-251 (1930) .
    • . . . Minute granules are liberated from the periplasmic sheath through budding and extrusion, which may multiply and may be transmissible 232425262728293031 . . .
    • . . . Their presence in syphilitic patients was regarded as confirmatory of the syphilitic nature of the lesions even in the absence of classical spiral forms 262730 . . .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
  28. Y Manouélian Syphilis tardive. Forms minuscules du Spirochaeta pallida. Spirochetogene syphilitique Ann Inst Pasteur 55, 698-708 (1935) .
    • . . . Minute granules are liberated from the periplasmic sheath through budding and extrusion, which may multiply and may be transmissible 232425262728293031 . . .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
  29. Y Manouélian Placentas syphilitiques, formes minuscules du tréponème et ultravirus syphilitique C rend Acad sc 200, 1439 (1935) .
    • . . . Minute granules are liberated from the periplasmic sheath through budding and extrusion, which may multiply and may be transmissible 232425262728293031 . . .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
  30. AS Warthin; RE Olsen The apparent sequence of spirochetes and granular forms in syphilitic buboes Am J Syphilis 15, 145 (1931) .
    • . . . Minute granules are liberated from the periplasmic sheath through budding and extrusion, which may multiply and may be transmissible 232425262728293031 . . .
    • . . . Their presence in syphilitic patients was regarded as confirmatory of the syphilitic nature of the lesions even in the absence of classical spiral forms 262730 . . .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
    • . . . Atypical and cystic forms of Treponema pallidum have been observed both extra- and intracellularly 30 . . .
    • . . . The intracellular localization of Treponema pallidum is another way of evading from destruction by the host immune system 3039 . . .
  31. LH Mattman Cell wall deficient forms: stealth pathogens , (1993) .
    • . . . Minute granules are liberated from the periplasmic sheath through budding and extrusion, which may multiply and may be transmissible 232425262728293031 . . .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
    • . . . Reconversion of cystic Borrelia burgdorferi into the typical spiral form has been demonstrated in vitro and in vivo 8103140. . . .
  32. E Klieneberger-Nobel The filterable forms of bacteria Bacteriol Rev 15, 77-103 (1951) .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
    • . . . The newly formed spirochetes are delicate L or metacyclic forms 253239. . . .
  33. H Noguchi A method for the pure cultivation of pathogenic Treponema pallidum J Exp Med 14, 99-112 (1911) .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
  34. P Seguin Treponema calligyrum et ultra-virus spirochétique C rend Soc biol 104, 247 (1930) .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
  35. A Sézary Les Formes atypiques et la Forme granuleuse du Tréponème pale Compt rend Soc biol 105, 444 (1930) .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
  36. P Lepine Forme visible et forme invisible du virus syphilitique Rev méd Par 48, 721 (1931) .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
  37. H Noguchi Spirochaetes Am J Syphilis 1, 261-346 (1917) .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
  38. AA Al-Qudah; A Mostratos; LB Quesnel A proposed life cycle for the Reiter treponeme J Appl Bacteriol 55, 417-428 (1983) .
    • . . . These spore-like minute granules (0.1–0.3 μm in diameter) may pass the 0.2 μm "China" filter (32) and can grow into young spirochetes 6192526272829303132333435363738 . . .
    • . . . This has also been observed in various types of spirochetes [e.g. 113870] including Borrelia burgdorferi 8 by transmission electron microscopy analyses . . .
  39. P Gastinel Precis de bacteriologie medicale. Collections de récis medicaux Paris , (1949) .
    • . . . The newly formed spirochetes are delicate L or metacyclic forms 253239. . . .
    • . . . Treponema pallidum may invade virtually all parenchymal and mesenchymal cells, including plasma cells, macrophages, neurons and glial cells 394043 . . .
    • . . . The intracellular localization of Treponema pallidum is another way of evading from destruction by the host immune system 3039 . . .
  40. NM Ovcinnikov; VV Delektorskij Current concepts of the morphology and biology of Treponema pallidum based on electron microscopy Br J Vener Dis 47, 315-328 (1971) .
    • . . . Reconversion of cystic Borrelia burgdorferi into the typical spiral form has been demonstrated in vitro and in vivo 8103140. . . .
    • . . . Treponema pallidum may invade virtually all parenchymal and mesenchymal cells, including plasma cells, macrophages, neurons and glial cells 394043 . . .
  41. G Steiner Morphologic appearances of spirochetal reproduction in tissues Arch Pathol 5, 189-199 (1940) .
    • . . . The occurrence of pleomorphic forms of Treponema pallidum in the brain in general paresis and their abundance in juvenile paresis is well documented 618264142. . . .
  42. C Levaditi; R Schoen Le cycle évolutif du "Treponema pallidum Bull acad méd (Paris) 98, 149-155 (1927) .
    • . . . The occurrence of pleomorphic forms of Treponema pallidum in the brain in general paresis and their abundance in juvenile paresis is well documented 618264142. . . .
  43. V Lauderdale; JN Goldman Serial ultrathin sectioning demonstrating the intracellularity of T. pallidum Br J Vener Dis 48, 87-96 (1972) .
    • . . . Treponema pallidum may invade virtually all parenchymal and mesenchymal cells, including plasma cells, macrophages, neurons and glial cells 394043 . . .
  44. JO Gebbers; HP Marder Unusual in vitro formation of cyst-like structures associated with human intestinal spirochaetosis Eur J Clin Microbiol Infect Dis 8, 302-306 (1989) .
    • . . . It has also been described in other spirochetal infections [e.g. 444546]. . . .
  45. J Blom; K Hovind-Hougen; HJ Jensen Electron microscopy of lymph nodes of hamsters experimentally infected with Treponema pertenue Acta Pathol Microbiol Scand [A] 85, 89-98 (1977) .
    • . . . It has also been described in other spirochetal infections [e.g. 444546]. . . .
  46. R Inada; Y Ido; R Hoki; Kanedo; H Ito The etiology and mode of infection and specific therapy of Weil's disease. [Spirochaeta icterohaemorrhagica] J Exp Med 23, 377-402 (1916) .
    • . . . It has also been described in other spirochetal infections [e.g. 444546]. . . .
  47. PH Duray; SR Yin; Y Ito; L Bezrukov; C Cox; MS Cho; W Fitzgerald; D Dorward; J Zimmerberg; L Margolis Invasion of human tissue ex vivo by Borrelia burgdorferi J Infect Dis 191, 1747-1754 (2005) .
    • . . . Their occurrence has been reported in skin lesions 14, in an ex vivo system in tonsil tissue 47 and on silver stained hippocampus section in a patient with concurrent Alzheimer disease (AD) and Lyme neuroborreliosis 48 . . .
  48. AB MacDonald Concurrent neocortical borreliosis and Alzheimer's disease: Demonstration of a spirochetal cyst form Ann New York Acad Sci 539, 468-470 (1988) .
    • . . . Their occurrence has been reported in skin lesions 14, in an ex vivo system in tonsil tissue 47 and on silver stained hippocampus section in a patient with concurrent Alzheimer disease (AD) and Lyme neuroborreliosis 48 . . .
  49. J Miklossy; A Kis; A Radenovic; L Miller; L Forro; R Martins; K Reiss; N Darbinian; P Darekar; L Mihaly; K Khalili Beta-amyloid deposition and Alzheimer's type changes induced by Borrelia spirochetes Neurobiol Aging 27, 228-236 (2006) .
    • . . . Intracellular localization of Borrelia burgdorferi was observed in macrophages and keratinocytes in the skin 14 and in neurons and glial cells in vitro and in vivo 3495051. . . .
    • . . . Recently we reported evidence that Borrelia burgdorferi can also persist in the brain in chronic Lyme neuroborreliosis and, in analogy to Treponema pallidum, may cause dementia, cortical atrophy and amyloid deposition 34951 . . .
  50. JA Livengood; RD Gilmore Invasion of human neuronal and glial cells by an infectious strain of Microbes Infect 8, 2832-2840 (2006) .
    • . . . Intracellular localization of Borrelia burgdorferi was observed in macrophages and keratinocytes in the skin 14 and in neurons and glial cells in vitro and in vivo 3495051. . . .
  51. J Miklossy Alzheimer's disease – A spirochetosis? Neuroreport 4, 841-848 (1993) .
    • . . . Intracellular localization of Borrelia burgdorferi was observed in macrophages and keratinocytes in the skin 14 and in neurons and glial cells in vitro and in vivo 3495051. . . .
    • . . . Borrelia burgdorferi spirochetes strains B31 and ADB1 3515253 were cultivated in BSK II medium 54 . . .
    • . . . Recently we reported evidence that Borrelia burgdorferi can also persist in the brain in chronic Lyme neuroborreliosis and, in analogy to Treponema pallidum, may cause dementia, cortical atrophy and amyloid deposition 34951 . . .
    • . . . Several observations suggested that Borrelia burgdorferi possesses amyloidogenic proteins 516869 . . .
    • . . . Like for Treponema pallidum in neurosyphilis, atypical and cystic forms of Borrelia burgdorferi were also observed intracellularly in the brains of these patients, as it has previously been documented 351 . . .
    • . . . That Borrelia burgdorferi was successfully cultivated from brains of the three patients with Lyme neuroborreliosis in BSK-II medium where pleomorphic and cystic forms were observed in the brain 351527273 suggests that at least part of the persisting spirochetes are viable . . .
    • . . . The accumulation of immunocompetent HLA-DR positive microglia and reactive astrocytes in the cerebral cortex of these patients clearly indicates the presence of chronic inflammation as previously suggested 51 . . .
  52. J Miklossy; S Kasas; RC Janzer; F Ardizzoni; H Loos Further morphological evidence for a spirochetal etiology of Alzheimer's Disease NeuroReport 5, 1201-1204 (1994) .
    • . . . Borrelia burgdorferi spirochetes strains B31 and ADB1 3515253 were cultivated in BSK II medium 54 . . .
    • . . . That Borrelia burgdorferi was successfully cultivated from brains of the three patients with Lyme neuroborreliosis in BSK-II medium where pleomorphic and cystic forms were observed in the brain 351527273 suggests that at least part of the persisting spirochetes are viable . . .
  53. J Miklossy; L Gern; P Darekar; RC Janzer; H Loos Senile plaques, neurofibrillary tangles and neuropil threads contain DNA? J Spirochetal and Tick-borne Dis 2, 1-5 (1995) .
    • . . . Borrelia burgdorferi spirochetes strains B31 and ADB1 3515253 were cultivated in BSK II medium 54 . . .
  54. BW Berger; MH Kaplan; IR Rothenberg; AG Barbour Isolation and characterization of the Lyme disease spirochete from the skin of patients with erythema chronicum migrans J Am Acad Dermatol 13, 444-449 (1985) .
    • . . . Borrelia burgdorferi spirochetes strains B31 and ADB1 3515253 were cultivated in BSK II medium 54 . . .
  55. E Aberer; PH Duray Morphology of Borrelia burgdorferi: structural patterns of cultured borreliae in relation to staining methods J Clin Microbiol 29, 764-72 (1991) .
    • . . . Spirochetes are known to bind Congo red and Thioflavin S 5556, both of which are widely used to stain amyloid . . .
    • . . . These results are in harmony with previous observations 85566 . . .
    • . . . Consequently, when the clinical and histopathologic features suggest syphilis or Lyme disease, the detection of these spirochetes in infected body fluids and tissues may be of diagnostic importance 5571. . . .
  56. D Scott; IR Siboo; EC Chan; A Klitorinos; R Siboo Binding of hemin and congo red by oral hemolytic spirochetes Oral Microbiol Immunol 8, 245-250 (1993) .
    • . . . Spirochetes are known to bind Congo red and Thioflavin S 5556, both of which are widely used to stain amyloid . . .
    • . . . They may act by binding to the outer sheath of Borrelia spirochetes [e.g. 56] . . .
  57. AD Zurn; F Mudry Conditions increasing the adrenergic properties of dissociated chick superior cervical ganglion neurons grown in long-term culture Dev Biol 117, 365-379 (1986) .
    • . . . Superior cervical ganglia from 8- to 12-day-old chicken embryos were dissociated as described previously 57 . . .
    • . . . The sympathetic neurons were then grown for 3–4 weeks in serum containing medium on a polyornithine substrate pre-coated with heart-conditioned medium 57 . . .
  58. H Fiumelli; M Kiraly; A Ambrus; P Magistretti; JL Martin Opposite regulation of calbindin and calretinin expression by brain-derived neurotrophic factor in cortical neurons J Neurochem 74, 1870-1877 (2000) .
    • . . . Neurons dissociated from the telencephalon of 21-day-old rat were cultured either on collagen or polylysine substrate in a serum-containing medium 58 . . .
  59. RC Janzer; MC Raff Astrocytes induce blood-brain barrier properties in endothelial cells Nature 325, 253-257 (1987) .
    • . . . Rat primary astrocytes (106) were prepared as described earlier 59 . . .
  60. J Miklossy; T Arai; JP Guo; A Klegeris; S Yu; EG McGeer; PL McGeer LRRK2 expression in normal and pathologic human brain and in human cell lines J Neuropathol Exp Neurol 65, 953-963 (2006) .
    • . . . The immunoreaction was revealed by diaminobenzidine (DAB) alone, or with nickel-ammonium sulfate as described previously 60 . . .
  61. J Miklossy; P Darekar; L Gern; RC Janzer; FT Bosman Bacterial peptidoglycan in neuritic plaques in Alzheimer's disease Azheimer's Res 2, 95-100 (1996) .
    • . . . A monoclonal antibody (Biogenesis 7263-1006 or Chemicon MAB995, dil.1: 200) for the analysis of the presence of bacterial peptidoglycan, a bacterial cell wall component of virtually all Eubacteria, including spirochetes, was also used as previously described in detail 61. . . .
  62. S Kasas; B Fellay; R Cargnello Observation of the action of Penicillin on Bacillus Subtilis using atomic force microscopy- technique for preparation of bacteria Surface Interface Anal 21, 400-407 (1994) .
    • . . . Samples were then stored at 4°C until used for the atomic force microscopy (AFM) analysis. 20–50 μl samples were put on the surface of a Nucleopore® filter of 2 μm hole size and were dried at room temperature in air, as previously described 62 . . .
  63. AC Pacheco e Silva Espirochetose dos centros nervos Memorias do hospicio de Juquery 0, 1-27 (1927) .
    • . . . Treponema pallidum persists in the brain and causes various neuropsychiatric disorders including dementia, cortical atrophy and amyloid deposition years or decades following the primary infection 636465 . . .
    • . . . The clinical and the pathological hallmarks of Alzheimer's disease, including beta-amyloid deposition are also present in the atrophic form of general paresis and in tertiary Lyme neuroborreliosis 363727374 . . .
  64. W Volland Die Kolloide Degeneration des Gehirns bei progressiver Paralyse in ihrer Beziehung zur lokalen Amyloidose Dtsch Path Gesellsch 31, 515-520 (1938) .
    • . . . Treponema pallidum persists in the brain and causes various neuropsychiatric disorders including dementia, cortical atrophy and amyloid deposition years or decades following the primary infection 636465 . . .
  65. H Noguchi; JW Moore A demonstration of Treponema pallidum in the brain of general paralysis cases J Exp Med 17, 232-238 (1913) .
    • . . . Treponema pallidum persists in the brain and causes various neuropsychiatric disorders including dementia, cortical atrophy and amyloid deposition years or decades following the primary infection 636465 . . .
  66. VP Mursic; G Wanner; S Reinhardt; B Wilske; U Busch; W Marget Formation and cultivation of Borrelia burgdorferi spheroplast-L-form variants Infection 24, 218-226 (1996) .
    • . . . These results are in harmony with previous observations 85566 . . .
  67. PH Hardy; EE Nell Influence of osmotic pressure on the morphology of the Reiter treponeme J Bacteriol 82, 967-978 (1961) .
  68. S Ohnishi; A Koide; S Koide Solution conformation and amyloid-like fibril formation of a polar peptide derived from a beta-hairpin in the OspA single-layer beta-sheet J Mol Biol 301, 477-489 (2000) .
    • . . . Several observations suggested that Borrelia burgdorferi possesses amyloidogenic proteins 516869 . . .
  69. S Ohnishi; A Koide; S Koide The roles of turn formation and cross-strand interactions in fibrillization of peptides derived from the OspA single-layer beta-sheet Protein Sci 10, 2083-2092 (2001) .
    • . . . Several observations suggested that Borrelia burgdorferi possesses amyloidogenic proteins 516869 . . .
  70. AE Ritchie Morphology of leptospires. The Biology of Parasitic Spirochetes , 19-37 (1976) .
    • . . . This has also been observed in various types of spirochetes [e.g. 113870] including Borrelia burgdorferi 8 by transmission electron microscopy analyses . . .
  71. ED Shapiro; MA Gerber Lyme disease Clin Infect Dis 31, 533-542 (2000) .
    • . . . Consequently, when the clinical and histopathologic features suggest syphilis or Lyme disease, the detection of these spirochetes in infected body fluids and tissues may be of diagnostic importance 5571. . . .
  72. J Miklossy Chronic inflammation and amyloidogenesis in Alzheimer's disease – role of spirochetes J Alzheimers Dis 13, 381-391 (2008) .
    • . . . That Borrelia burgdorferi was successfully cultivated from brains of the three patients with Lyme neuroborreliosis in BSK-II medium where pleomorphic and cystic forms were observed in the brain 351527273 suggests that at least part of the persisting spirochetes are viable . . .
    • . . . The clinical and the pathological hallmarks of Alzheimer's disease, including beta-amyloid deposition are also present in the atrophic form of general paresis and in tertiary Lyme neuroborreliosis 363727374 . . .
  73. J Miklossy Biology and neuropathology of dementia in syphilis and Lyme disease Handbook of Clinical Neurology 89, 825-844 (2008) .
    • . . . That Borrelia burgdorferi was successfully cultivated from brains of the three patients with Lyme neuroborreliosis in BSK-II medium where pleomorphic and cystic forms were observed in the brain 351527273 suggests that at least part of the persisting spirochetes are viable . . .
    • . . . The vegetative and cystic forms including the vesicular blubs and free vesicular structures of Borrelia burgdorferi all contain the biologically active spirochetal surface proteins indicating that they all elicit inflammatory responses including complement activation [reviewed in 73]. . . .
    • . . . The clinical and the pathological hallmarks of Alzheimer's disease, including beta-amyloid deposition are also present in the atrophic form of general paresis and in tertiary Lyme neuroborreliosis 363727374 . . .
  74. J Miklossy; S Rosemberg; PL McGeer Beta amyloid deposition in the atrophic form of general paresis Alzheimer's Disease: New advances. Medimond. Proceedings of the 10th International Congress on Alzheimer's Disease , 429-433 (2006) .
    • . . . The clinical and the pathological hallmarks of Alzheimer's disease, including beta-amyloid deposition are also present in the atrophic form of general paresis and in tertiary Lyme neuroborreliosis 363727374 . . .
Expand