G-CSF induces E-selectin ligand expression on human myeloid cells.
Clinical use of G-CSF can result in vascular and inflammatory complications. To investigate the molecular basis of these effects, we analyzed the adherence of G-CSF-mobilized human peripheral blood leukocytes (ML) to inflamed (TNF-alpha-stimulated) vascular endothelium. Studies using parallel plate assays under physiologic flow conditions and intravital microscopy in a mouse inflammation model each showed that ML take part in heightened adhesive interactions with endothelium compared to unmobilized (native) blood leukocytes, mediated by markedly increased E-selectin receptor-ligand interactions. Biochemical studies showed that ML express the potent E-selectin ligand HCELL (ref. 8) and another, previously unrecognized approximately 65-kDa E-selectin ligand, and possess enhanced levels of transcripts encoding glycosyltransferases (ST3GalIV, FucT-IV and FucT-VII) conferring glycan modifications associated with E-selectin ligand activity. Enzymatic treatments and physiologic binding assays showed that HCELL and the approximately 65-kDa E-selectin ligand contribute prominently to the observed G-CSF-induced myeloid cell adhesion to inflamed endothelium. Treatment of normal human bone marrow cells with a pharmacokinetically relevant concentration of G-CSF in vitro resulted in increased expression of these two molecules, coincident with increased transcripts encoding pertinent glycosyltransferases and heightened E-selectin binding. These findings provide direct evidence for a role of G-CSF in the induction of E-selectin ligands on myeloid cells, thus providing mechanistic insight into the pathobiology of G-CSF complications.
Dagia
Nilesh M
NM
Department of Dermatology, Brigham & Women's Hospital, 77 Avenue Louis Pasteur, Room 671, Harvard Medical School, Boston, Massachusetts 02115, USA.
Gadhoum
Samah Z
SZ
Knoblauch
Christine A
CA
Spencer
Joel A
JA
Zamiri
Parisa
P
Lin
Charles P
CP
Sackstein
Robert
R
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D019040:E-Selectin / Q000378:metabolism*; D005647:Fucosyltransferases / Q000378:metabolism; D005786:Gene Expression Regulation; D016179:Granulocyte Colony-Stimulating Factor / Q000378:metabolism*; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism*; D016256:Lewis X Antigen; D051379:Mice; D022423:Myeloid Cells / Q000166:cytology*; D012799:Sialyltransferases / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase
D019040:E-Selectin; D005647:Fucosyltransferases; D016179:Granulocyte Colony-Stimulating Factor; D018960:Hyaluronan Receptors; D016256:Lewis X Antigen; D012799:Sialyltransferases; D014409:Tumor Necrosis Factor-alpha; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase; C066693:FUT4 protein, human; C502597:FUT7 protein, human; C497483:CD44 protein, human