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Formation and cultivation ofBorrelia burgdorferi spheroplast-L-form variants

Sphäroplasten-L Phase-Varianten vonBorrelia burgdorferi: Morphologische Analyse und Kultivierung

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Summary

As clinical persistence ofBorrelia burgdorferi in patients with active Lyme borreliosis occurs despite obviously adequate antibiotic therapy,in vitro investigations of morphological variants and atypical forms ofB. burgdorferi were undertaken. In an attempt to learn more about the variation ofB. burgdorferi and the role of atypical forms in Lyme borreliosis, borreliae isolated from antibiotically treated and untreated patients with the clinical diagnosis of definite and probable Lyme borreliosis and from patient specimens contaminated with bacteria were investigated. Furthermore, the degeneration of the isolates during exposure to penicillin Gin vitro was analysed. Morphological analysis by darkfield microscopy and scanning electron microscopy revealed diverse alterations. Persisters isolated from a great number of patients (60–80%) after treatment with antibiotics had an atypical form. The morphological alterations in culture with penicillin G developed gradually and increased with duration of incubation. Pleomorphism, the presence of elongated forms and spherical structures, the inability of cells to replicate, the long period of adaptation to growth in MKP-medium and the mycoplasma-like colonies after growth in solid medium (PMR agar) suggest thatB. burgdorferi produce spheroplast-L-form variants. With regard to the polyphase course of Lyme borreliosis, these forms without cell walls can be a possible reason whyBorrelia survive in the organism for a long time and the cell-wall-dependent antibody titers disappear and emerge after reversion.

Zusammenfassung

Mit Hilfe von Dunkelfeld- und elektronenmikroskopischer Darstellungen wurden Borrelien-Stämme auf morphologische Varianten untersucht. Es interessierte das Vorkommen und die Bedeutung von atypischen Formen und Sphäroplasten bei antibiotisch behandelten und unbehandelten Patienten mit der klinisch diagnostizierten aktiven Lyme Borreliose bzw. beim Verdacht auf Lyme Borreliose. Ferner untersuchten wir die Zellveränderungen unter Penicillin G-Wirkungin vitro. Die morphologische Analyse zeigte neben der helikalen Form sehr heterogene Veränderungen unter den Borrelien. Die morphologische Veränderung unter Penicillineinwirkung war stark von der Einwirkungszeit abhängig. Die Borrelien-Persister, isoliert von Patienten nach Antibiotikatherapie, zeigten jeweils in großer Zahl (60–80%) atypische Formen. Pleomorphismus, elongierte Zellen und sphäroide Strukturen, Bewegungs- und Vermehrungsunfähigkeit, schlechtes Wachstum im MKP-Medium, sowie Mykoplasmaähnliche Kolonien auf PMR-Agar lassen beiBorrelia burgdorferi die Bildung von Sphäroplasten-L Phase-Varianten vermuten. In Hinblick auf den Krankheitsverlauf der Lyme Borreliose wären diese zellwandfreien Formen eine Möglichkeit für die Borrelien, längere Zeit im Organismus zu überdauern. Die zellwandabhängigen Antikörper können so verschwinden und wieder auftauchen, wenn die Organismen revertieren.

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Authors and Affiliations

  1. Dipl. Biol., Max von Pettenkofer-Institut, Ludwig-Maximilians-Universität München, Pettenkoferstr. 9a, D-80336, München

    Vera Preac Mursic Ph. D., Sylvia Reinhardt, Bettina Wilske M. D. & U. Busch

  2. Botanisches Institut, Universität München, Menzinger Str. 67, D-80638, München

    G. Wanner Ph. D.

  3. Kinderklinik, Universität München, Lindwurmstr. 4, D-80337, München, Germany

    W. Marget

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Mursic, V.P., Reinhardt, S., Wilske, B. et al. Formation and cultivation ofBorrelia burgdorferi spheroplast-L-form variants. Infection 24, 218–226 (1996). https://doi.org/10.1007/BF01781096

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