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Tick factors andin vitro cultivation influence the protein profile, antigenicity and pathogenicity of a clonedBorrelia garinii isolate fromIxodes ricinus hemolymph

Faktoren der Zecke undin vitro-Kultivierung beeinflussen das Proteinmuster, die Antigenität und die Pathogenität eines aus Hämolymphe gewonnenen kloniertenBorrelia garinii Isolats

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Summary

ABorrelia garinii isolate (NE11H) was obtained from the hemolymph of infedIxodes ricinus. NE11H expressed four major proteins of 33 kDa, 32 kDa, 23 kDa and 22 kDa. Duringin vitro culture, NE11H successively lost the expression of the 22 kDa and 23 kDa proteins and the NE11H variant (NE11Hp15) was not recognized by an immune serum specific for the OspC protein (anti-OspC IS). However, when reintroduced into tick midguts, NE11Hp15 spirochetes present in the midgut again reacted with anti-OspC IS. A clone derived from the wild type line, cNE11H, lacked the 22 kDa but not the 23 kDa protein. The 23 kDa protein of cNE11H was recognized by anti-OspC IS. In addition, the two descendant lines (NE11Hp15 and cNE11H) lost their capacity to induce clinical arthritis in SCID mice. When cNE11H was reintroduced into ticks and reisolated from various tick organs, most reisolates presented the same reaction with anti-OspC IS as cNE11H. Interestingly, two reisolates obtained from the tick midgut reexpressed large amounts of the 22 kDa protein which was recognized by anti-OspC IS and these two reisolates induced clinical arthritis in SCID mice. The results confirm that proteins of 22/23 kDa are differentially expressed duringin vitro subcultures and in ticks, and show that proteins which are not detectable afterin vitro culture may be reexpressed after reexposure ofB. burgdorferi to its former environment in the tick. The data suggest that the pathogenicity ofB. burgdorferi for mice might be influenced by environmental factors via differential expression of 22/23 kDa proteins.

Zusammenfassung

EinBorrelia garinii Isolat (NE11H) wurde aus der Hämolymphe nicht gefütterterIxodes ricinus Zecken gewonnen. NE11H exprimierte vier Hauptproteine mit einem Molekulargewicht von jeweils 33, 32, 23 und 22 kDa. Im Laufe derin vitro Kultur entstand eine Variante (NE11Hp15), die durch den Verlust der 22 und 23 kDa Proteine gekennzeichnet war. NE11Hp15 zeigte keine Reaktion mit einem Immunserum gegen OspC. Wurde NE11Hp15 in Zecken passagiert, fand sich wieder eine Reaktion mit dem anti-OspC-Immunserum. Ein Klon der Ausgangslinie NE11H (cNE11H) war durch das Fehlen des 22 kDa, nicht jedoch des 23 kDa Proteins, gekennzeichnet. Das 23 kDa Protein von cNE11H wurde durch anti-OspC-Immunserum erkannt. Sowohl NE11Hp15 als auch cNE11H waren nicht in der Lage, eine klinisch nachweisbare Arthritis in SCID-Mäuse zu induzieren. Nach Reinokulation von cNE11H in Zecken und anschließender Reisolation aus verschiedenen Zeckenorganen, zeigten die meisten Reisolate eine Reaktion mit anti-OspC-Immunserum wie der Ausgangsklon cNE11H. Interessanterweise fand sich bei zwei Reisolaten aus dem Zeckenmitteldarm eine starke Expression des 22 kDa Proteins, welches durch das anti-OspC-Immunserum erkannt wurde. Darüberhinaus induzierten diese beiden Reisolate eine klinisch nachweisbare Arthritis in SCID-Mäusen. Zusammengefaßt zeigen unsere Resultate, daß die spirochätalen 22/23 kDa Proteine während der Zeckenpassage differentiell exprimiert werden. Darüberhinaus zeigen sie auch, daß spirochätale Proteine, die nach Kultivierungin vitro nicht mehr nachweisbar sind, dann reexprimiert werden können, wennB. burgdorferi der Mikroumgebung des Zeckenorganismus ausgesetzt wird. Wir postulieren, daß die Pathogenität vonB. burgdorferi in Mäusen durch umgebungsbedingte differentielle Expression der 22 und 23 kDa Proteine beeinflußt werden könnte.

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Authors and Affiliations

  1. Institut de Zoologie, Université de Neuchâtel, Neuchâtel, Switzerland

    Chang Min Hu &  Lise Gern

  2. Max-Planck-Institut für Immunbiologie, D-79108, Freiburg

    M. Simon

  3. Institut für Immunologie, Ruprecht-Karls-Universität, D-69120, Heidelberg, Germany

    M. D. Kramer

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Hu, C.M., Gern, L., Simon, M. et al. Tick factors andin vitro cultivation influence the protein profile, antigenicity and pathogenicity of a clonedBorrelia garinii isolate fromIxodes ricinus hemolymph. Infection 24, 251–257 (1996). https://doi.org/10.1007/BF01781105

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