Internalization of monomeric lipopolysaccharide occurs after transfer out of cell surface CD14

被引:65
作者
Vasselon, T
Hailman, E
Thieringer, R
Detmers, PA
机构
[1] Merck & Co Inc, Merck Sharp & Dohme Res Labs, Dept Endocrinol & Chem Biol, Rahway, NJ 07065 USA
[2] Washington Univ, Sch Med, Div Lab Med, St Louis, MO 63110 USA
关键词
enhanced green fluorescent protein; U373; cells; intracellular trafficking;
D O I
10.1084/jem.190.4.509
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Lipopolysaccharide (LPS) fluorescently labeled with boron dipyrromethane (BODIPY) first binds to the plasma membrane of CD 14-expressing cells and is subsequently internalized. Intracellular LPS appears in small vesicles near the cell surface and later in larger, punctate structures identified as the Golgi apparatus. To determine if membrane (m)CD14 directs the movement of LPS to the Golgi apparatus, an mCD14 chimera containing enhanced green fluorescent protein (mCD14-EGFP) was used to follow trafficking of mCD14 and BODIPY-LPS in stable transfectants. The chimera was expressed strongly on the cell surface and also in a Golgi complex-like structure, mCD14-EGFP was functional in mediating binding of and responses to LPS. BODIPY-LPS presented to the transfectants as complexes with soluble CD14 first colocalized with mCD14-EGFP on the cell surface. However, within 5-10 mn, the BODIPY-LPS distributed to intracellular vesicles that did not contain mCD14-EGFP, indicating that mCD14 did not accompany LPS during endocytic movement. These results suggest that monomeric LPS is transferred out of mCD14 at the plasma membrane and traffics within the cell independently of mCD14. In contrast, aggregates of LPS were internalized in association with mCD14, suggesting that LPS clearance occurs via a pathway distinct from that which leads to signaling via monomeric LPS.
引用
收藏
页码:509 / 521
页数:13
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