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J. Biol. Chem., Vol. 278, Issue 36, 33645-33653, September 5, 2003

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Acylated Cholesteryl Galactoside as a Novel Immunogenic Motif in Borrelia burgdorferi Sensu Stricto^*

Nicolas W. J. Schröder , Ursula Schombel , Holger Heine ¶, Ulf B. Göbel , Ulrich Zähringer  and Ralf R. Schumann  ||

From the Institut für Mikrobiologie und Hygiene, Universitätsklinikum "Charité," Medizinische Fakultät der Humboldt-Universität zu Berlin, Dorotheenstrasse 96, D-10117 Berlin, Germany and the Division of Immunochemistry and ^¶Junior Research Group Innate Immunity, Research Center Borstel, Liebniz Center for Medicine and Biosciences, Parkallee 22, D-23845 Borstel, Germany

Borrelia burgdorferi sensu lato is the causing agent of Lyme disease, an infectious disease frequently occurring in the United States, Europe, and Northern Asia. Currently, diagnosis of and vaccination strategies against this pathogen are exclusively based on proteinaceous structures. Here we report on a novel class of immunogenic glycolipids purified from B. burgdorferi sensu stricto B31. Employing a butanol/water extraction procedure with subsequent Bligh/Dyer extraction of the organic phase, thin layer chromatography analysis revealed the presence of three distinct glycolipids, which were chemically analyzed employing combined gas-liquid chromatography/mass spectroscopy, matrix-assisted laser desorption/ionization mass spectrometry, and NMR. We identified acylated cholesteryl galactoside (ACG) next to cholesteryl galactoside and -monogalactosyl-diacylglycerol. After extensive purification, the glycolipids investigated failed to cause proinflammatory responses in human cells transfected with human toll-like receptor (TLR)-2 or -4. However, we observed a marked recognition of ACG by sera derived from patients suffering from Lyme disease. These data indicate that newly described ACG is involved in developing host immunity during Lyme disease and thus may be useful for diagnosis and vaccination.

Received for publication, June 3, 2003 , and in revised form, June 16, 2003.

Note Added in Proof—During the review process of this manuscript, an article has appeared confirming the chemical part of our findings (Ben-Menachem, G., Kubler-Kielb, J., Coxon, B., Yergey, A., and Schnerson, R. (2003) Proc. Natl. Acad. Sci. U. S. A. 100, 7913–7918).

^* This work was supported in part by Deutsche Forschungsgemeinschaft Grants Schr 726/1-1 (to R. R. S. and N. W. J. S.), ZA 149/5-1 (to U. Z.), and He 2758/3-1 (to H. H.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence and reprint requests should be addressed. Tel.: 49-30-450-524141; Fax: 49-30-450-524904; E-mail: ralf.schumann{at}charite.de.

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