Changes in subcellular localization and surface expression of L-selectin, alkaline phosphatase, and Mac-1 in human neutrophils during stimulation with inflammatory mediators.
The localization of the adhesion protein L-selectin in human neutrophils was determined by subcellular fractionation and immunoelectron microscopy and compared with the localization of Mac-1 (alpha m beta 2) and alkaline phosphatase, the marker for secretory vesicles. L-selectin was found to be localized exclusively on the plasma membrane of unstimulated cells and also of stimulated cells, although markedly diminished. This was in contrast to Mac-1, which was also localized in secretory vesicles and in specific/gelatinase granules as shown previously [Sengeløv, H., et al. J. Clin. Invest. (1993) 92, 1467-1476]. Stimulation of neutrophils with inflammatory mediators such as tumor necrosis factor (TNF), platelet-activating factor (PAF), or f-Met-Leu-Phe (fMLP), induced parallel up-regulation of the surface membrane content of alkaline phosphatase and Mac-1 and down-regulation of L-selectin, as evidenced by flow cytometry. Preimbedding immunoelectron microscopy confirmed that L-selectin was present mainly on tips of microvilli in unstimulated cells and showed that alkaline phosphatase and Mac-1 were randomly distributed on the surface membrane of fMLP-stimulated cells. These studies indicate that the transition of neutrophils from L-selectin-presenting cells to Mac-1-presenting cells induced by inflammatory mediators is mediated by incorporation of secretory vesicle membrane, rich in Mac-1 and devoid of L-selectin, into the plasma membrane.
Borregaard
N
N
Department of Hematology, University Hospital, Rigshospitalet, Copenhagen, Denmark.
Kjeldsen
L
L
Sengeløv
H
H
Diamond
M S
MS
Springer
T A
TA
Anderson
H C
HC
Kishimoto
T K
TK
Bainton
D F
DF
D000469:Alkaline Phosphatase / Q000097:blood*; D015815:Cell Adhesion Molecules / Q000097:blood*; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000648:ultrastructure; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D019041:L-Selectin; D016177:Macrophage-1 Antigen / Q000097:blood*; D016253:Microscopy, Immunoelectron; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D009504:Neutrophils / Q000737:chemistry* / Q000166:cytology / Q000648:ultrastructure; D010972:Platelet Activating Factor / Q000494:pharmacology*; D013347:Subcellular Fractions; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology
D015815:Cell Adhesion Molecules; D016177:Macrophage-1 Antigen; D010972:Platelet Activating Factor; D014409:Tumor Necrosis Factor-alpha; D019041:L-Selectin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D000469:Alkaline Phosphatase