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by Blocking the Binding of LPS to CD14+ Cells1
*
Department of Biochemistry, Juntendo University School of Medicine, Tokyo, Japan;
Research Division of Innate Immunity, Matsuzono Pharmacy, Iwate, Japan;
Laboratory of Immunopharmacology of Microbial Products, Tokyo University of Pharmacy and Life Science School of Pharmacy, Tokyo, Japan;
Central Research Laboratories, Seikagaku Corporation, Tokyo, Japan; and
¶
Division of Infectious Diseases, Centre Hospitalier Universitaire Vaudois-Lausanne, Lausanne, Switzerland
Mammalian myeloid and epithelial cells express several kinds of
antibacterial peptides (
-/
-defensins and cathelicidins) that
contribute to the innate host defense by killing invading
micro-organisms. In this study we evaluated the LPS-neutralizing
activities of cathelicidin peptides human CAP18 (cationic antibacterial
proteins of 18 kDa) and guinea pig CAP11 using the CD14+
murine macrophage cell line RAW264.7 and the murine endotoxin shock
model. Flow cytometric analysis revealed that CAP18 and CAP11 inhibited
the binding of FITC-conjugated LPS to RAW264.7 cells. Likewise,
Northern and Western blot analyses indicated that CAP18 and CAP11
suppressed LPS-induced TNF- mRNA and protein expression by RAW264.7
cells. Interestingly, CAP18 and CAP11 possessed LPS-binding activities,
and they strongly suppressed the interaction of LPS with LPS binding
protein that mediates the transport of LPS to CD14 to facilitate the
activation of CD14+ cells by LPS. Moreover, when CAP18 and
CAP11 were preincubated with RAW264.7 cells, they bound to the cell
surface CD14 and inhibited the binding of FITC-LPS to the cells.
Furthermore, in the murine endotoxin shock model, CAP18 or CAP11
administration inhibited the binding of LPS to CD14+ cells
(peritoneal macrophages) and suppressed LPS-induced TNF- expression
by these cells. Together these observations indicate that cathelicidin
peptides CAP18 and CAP11 probably exert protective actions against
endotoxin shock by blocking the binding of LPS to CD14+
cells, thereby suppressing the production of cytokines by these cells
via their potent binding activities for LPS and
CD14.
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