title: In vitro effects of oxidized low density lipoprotein on CD11b/CD18 and L-selectin presentation on neutrophils and monocytes with relevance for the in vivo situation.
abstract: Oxidized LDL (oxLDL) has been identified as a potent stimulus of leukocyte adhesion to endothelium, a hallmark of early atherogenesis. A cytofluorometric study was performed to further characterize the mechanisms by which oxLDL stimulates the rapid adhesion of leukocytes to endothelium in vitro and in vivo. Incubation (30 minutes at 37 C) of whole blood (diluted with buffered saline to 1 x 10(6) leukocytes/ml) with oxLDL (0.85 mg LDL cholesterol/ml; oxidized by 7.5 mumol/L Cu2+ for 18 hours) but not native LDL stimulated the upregulation of CD11b/CD18 adhesion receptors on neutrophils (anti-leu-15 binding: 178 +/- 16% of baseline, P < 0.01, means +/- SD of n = 10 experiments) and on monocytes (169 +/- 34% of baseline, P < 0.01). This phenomenon was almost entirely inhibited by n-butanol or the vasoactive drug pentoxifylline (PTX), which also significantly reduced oxLDL-induced leukocyte adhesion to venular and arteriolar endothelium, as assessed by intravital microscopy on the dorsal skinfold chamber in hamsters (venules: 49 +/- 19 versus 120 +/- 34 cells/mm2, P < 0.05; arterioles: 9 +/- 4 versus 52 +/- 7 cells/mm2, P < 0.01) 30 minutes after intravenous injection of oxLDL (4 mg/kg body weight; means +/- SD of n = 7 hamsters per group). Butanol and PTX also significantly reduced the upregulation of CD11b/CD18 by f-methionyl-leucyl-phenylalanine (fMLP) and platelet-activating factor (PAF) but not by phorbol myristate acetate (PMA). Whereas fMLP and PAF stimulate leukocytes via binding to specific cell surface receptors and triggering complex signal transduction pathways, PMA bypasses these pathways and directly activates intracellular protein kinase C. By analogy, we propose that oxLDL upregulates CD11b/CD18 through its previously documented ability to stimulate the generation of second messengers. The effect of n-butanol and PTX on receptor presentation cannot be explained by changes in plasma membrane fluidity, as both agents failed to reverse the decrease in plasma membrane fluidity of neutrophils after stimulation with oxLDL, as assessed by fluorescence anisotropy measurement of the membrane marker diphenylhexatriene. Incubation of isolated neutrophils but not of whole blood with oxLDL resulted in a significant loss of L-selectin from the neutrophil surface (anti-TQ-1 binding: 40 +/- 13% of baseline, P < 0.01). A significant loss of this adhesion receptor on neutrophils and monocytes was also observed after stimulation of isolated neutrophils and whole blood with fMLP, PAF, and PMA.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Institute for Surgical Research, University of Munich, Germany.
authors.forename: H A
authors.initials: HA
authors.lastname: Lehr
authors.forename: F
authors.initials: F
authors.lastname: Krombach
authors.forename: S
authors.initials: S
authors.lastname: Münzing
authors.forename: R
authors.initials: R
authors.lastname: Bodlaj
authors.forename: S I
authors.initials: SI
authors.lastname: Glaubitt
authors.forename: D
authors.initials: D
authors.lastname: Seiffge
authors.forename: C
authors.initials: C
authors.lastname: Hübner
authors.forename: U H
authors.initials: UH
authors.lastname: von Andrian
authors.forename: K
authors.initials: K
authors.lastname: Messmer
mesh_terms: D020001:1-Butanol; D000328:Adult; D000818:Animals; D000440:Butanols / Q000494:pharmacology; D018821:CD18 Antigens / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002478:Cells, Cultured; D006224:Cricetinae; D005260:Female; D005434:Flow Cytometry; D005454:Fluorescence Polarization; D006801:Humans; D019041:L-Selectin; D008077:Lipoproteins, LDL / Q000737:chemistry / Q000502:physiology*; D016177:Macrophage-1 Antigen / Q000378:metabolism*; D008297:Male; D008647:Mesocricetus; D009000:Monocytes / Q000502:physiology*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000502:physiology*; D010431:Pentoxifylline / Q000494:pharmacology; D010972:Platelet Activating Factor / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D000440:Butanols; D018821:CD18 Antigens; D015815:Cell Adhesion Molecules; D008077:Lipoproteins, LDL; D016177:Macrophage-1 Antigen; D010972:Platelet Activating Factor; D019041:L-Selectin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D020001:1-Butanol; D013755:Tetradecanoylphorbol Acetate; D010431:Pentoxifylline