title: The inhibitory effect of dexamethasone on lymphocyte adhesion molecule expression and intercellular aggregation is not mediated by lipocortin 1.
abstract: Glucocorticoids exert their anti-inflammatory activity through multiple pathways which include the inhibition of cell adhesion events. The glucocorticoid-induced protein lipocortin 1 (LC1) has reported anti-inflammatory properties and has been proposed as a putative mediator of the anti-inflammatory effects of glucocorticoids. The role of LC1 in mediating the glucocorticoid inhibition of lymphocyte adhesion and cell adhesion molecule (CAM) expression was investigated in vitro using a microaggregation assay, flow cytometry and confocal microscopy. Lymphocytes stimulated for 96 h with plastic-bound OKT3 antibody showed significant increases in LFA-1 and CD2 expression. Dexamethasone (DEX; 10(-6) M) inhibited this increase but the neutralizing anti-LC1 MoAb 1A (5 microg/ml) failed to reverse the DEX effect; neither was purified human LC1 (50 x 10(-9) M) able to inhibit CAM expression. The biological activity of the LC1 was confirmed by its ability to suppress monocyte phagocytosis and respiratory burst in response to bovine serum albumin (BSA)-anti-BSA complexes. OKT3 stimulation of cultured mononuclear cells resulted in intercellular aggregation, scored microscopically using a visual index. This aggregation was completely reversed by 10-6 M DEX but unaffected by LC1 (50 x 10(-9) M). Significant intracellular expression of lymphocyte LC1 was observed using the anti-LC1 MoAb 1B in saponin-permeabilized cells. Distribution of LC1 had a diffuse, cytoplasmic pattern. LC1 expression was reduced following 3 h treatment with 10(-6) M DEX. These findings indicate that the DEX effects on lymphocyte adhesion and CAM expression are not mediated by LC1. Thus the reported in vivo effects of LC1 on leucocyte adhesion and transmigration probably occur through functional/conformation changes of surface CAM, rather than by alteration in expression.
authors.affiliation: Arthritis Research Section, Department of Biochemical Pharmacology, St Bartholomew's & the Royal London School of Medicine and Dentistry, University of London, London, UK. n.j.goulding@mds.qmw.ac.uk
authors.forename: N J
authors.initials: NJ
authors.lastname: Goulding
authors.forename: S
authors.initials: S
authors.lastname: Ogbourn
authors.forename: N
authors.initials: N
authors.lastname: Pipitone
authors.forename: P
authors.initials: P
authors.lastname: Biagini
authors.forename: R
authors.initials: R
authors.lastname: Gerli
authors.forename: C
authors.initials: C
authors.lastname: Pitzalis
mesh_terms: D017305:Annexin A1 / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000494:pharmacology / Q000502:physiology*; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000936:Antigen-Antibody Complex / Q000378:metabolism; D001667:Binding, Competitive; D018801:CD2 Antigens / Q000096:biosynthesis*; D002449:Cell Aggregation / Q000187:drug effects; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D016169:Lymphocyte Function-Associated Antigen-1 / Q000096:biosynthesis*; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D009000:Monocytes / Q000166:cytology / Q000187:drug effects; D010587:Phagocytosis / Q000187:drug effects
chemical_list: D017305:Annexin A1; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D018801:CD2 Antigens; D005938:Glucocorticoids; D016169:Lymphocyte Function-Associated Antigen-1; D003907:Dexamethasone