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001-es BibID:	BIBFORM068948
Első szerző:	Antal-Szalmás Péter (laboratóriumi szakorvos)
Cím:	Diverging Pathways for Lipopolysaccharide and CD14 in Human Monocytes / Peter Antal-Szalmas, Miriam J. J. G. Poppelier, Roel Broekhuizen, Jan Verhoef, Jos A. G. van Strijp, Kok P. M. van Kessel
Dátum:	2000
ISSN:	0196-4763
Megjegyzések:	Background: CD14 is considered to be the major endotoxin(lipopolysaccharide [LPS]) binding molecule on humanmonocytes. It initiates cellular response, but its rolein the clearance of LPS is not well understood. Underconditions that ensure totally CD14-dependent LPS bindingon human monocytes, the internalization mechanismsof LPS and CD14 were studied.Methods: The uptake and intracellular distribution offluorescein isothiocyanate (FITC)-LPS and CD14 was determinedby flow cytometry, trypan blue quenching, andconfocal fluorescence microscopy. Incubation of surfacebiotinylatedcells with LPS at 37?C or 4?C and subsequentsubfractionation was used to further characterize CD14internalization. The amount of the intracellular CD14 wasestimated by CD14 enzyme-linked immunosorbent assay(ELISA).Results: The internalization rate of 10 ng/ml FITC-LPSwith 1% human serum was 1% of bound endotoxin perminute, whereas CD14 expression did not decrease at thesame time surface. We proved the presence of an intracellularCD14 pool (2.68 3 106 molecules per unstimulatedmonocyte) and could show that internalized FITCLPSmolecules can be found in different intracellularcompartments than CD14. Subfractionation of LPS-treatedbiotinylated monocytes showed no change in biotinylatedCD14 in the membrane fraction independently of theincubation temperature (37?C or at 4?C) used, indicatingthat these CD14 molecules were not taken up by an activeprocess.Conclusions: These data indicate the presence of a largeintracellular CD14 pool in monocytes with a yet unknownfunction, and suggest that LPS and CD14 molecules can beinternalized independently after association on the cellsurface.
Tárgyszavak:	Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban LPS internalization CD14
Megjelenés:	Cytometry 41 (2000), p. 279-288. -
További szerzők:	Poppelier, Miriam J. J. G Broekhuizen, Roel Verhoef, Jan Strijp, Jos A. G., van Kessel, Kok P. M., van
Internet cím:	Intézményi repozitóriumban (DEA) tárolt változat
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