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*
Institut für Mikrobiologie und Hygiene, Universitätsklinikum Charité Medizinische Fakultät der Humboldt-Universität zu Berlin, Berlin, Germany; and
Laborgruppe Immunchemie, Forschungszentrum Borstel, Borstel, Germany
Culture supernatants from Treponema maltophilum
associated with periodontitis in humans and Treponema
brennaborense found in a bovine cattle disease accompanied with
cachexia caused a dose-dependent TNF-
synthesis in human monocytes
increasing with culture time. This activity could be reduced
significantly by blocking the CD14-part of the LPS receptor using the
My 4 mAb and by polymyxin B. In the murine macrophage cell line RAW
264.7, Treponema culture supernatants induced TNF-
secretion in a LPS binding protein (LBP)-dependent fashion. To enrich
for active compounds, supernatants were extracted with butanol, while
whole cells were extracted using a phenol/water method resulting in
recovery of material exhibiting a similar activity profile. An LPS-LBP
binding competition assay revealed an interaction of the treponeme
phenol/water extracts with LBP, while precipitation studies implied an
affinity to polymyxin B and endotoxin neutralizing protein. Macrophages
obtained from C3H/HeJ mice carrying a Toll-like receptor (TLR)-4
mutation were stimulated with treponeme extracts for NO release to
assess the role of TLRs in cell activation. Furthermore, NF-
B
translocation in TLR-2-negative Chinese hamster ovary (CHO) cells was
studied. We found that phenol/water-extracts of the two strains use
TLRs differently with T. brennaborense-stimulating cells
in a TLR-4-dependent fashion, while T.
maltophilum-mediated activation apparently involved TLR-2.
These results indicate the presence of a novel class of glycolipids in
Treponema initiating inflammatory responses involving
LBP, CD14, and TLRs.
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