Modulation of L-selectin expression by chemotactic factors and a cytokine, GM-CSF

Abstract

L-selectin is an adhesion molecule with constitutive expression located on the membrane of granulocytes, monocytes and lymphocytes. It is involved in the early stages of migration of these cells toward either the sites of inflammation or lymphoid tissues. After the cells are activated, Lselectin is down-regulated with shedding of a soluble fragment. Flow cytometry was used to measure L-selectin expression levels on the granulocyte surface, after incubation with a phorbol esther (PMA), two chemotactic factors (fMLP and LTB4) and a cytokine (GM-CSF). Under basal conditions, the expression of L-selectin was found in a high percentage (95.0 ± 0.7) of granulocytes; PMA stimulation led to a marked decrease in expression (3.2 ± 0.6). Chemotactic factors also led to a significant decrease in L-selectin expression (69.9 ± 5.0 for LTB4, and 53.70 ± 4.3 for fMLP), whereas the incubation with GM-CSF produced no significant changes (89.1 ± 4.8). When all the conditions were compared, the PMA effect was significantly higher than those observed with other stimuli; furthermore, the expression upon incubation with fMLP and LTB4 was statistically significant. These results suggest that the level of activation reached by granulocytes is directly related to their capacity for shedding L-selectin from the cell surface, and that these levels are lowered after the stimulation by chemotactic factors. GM-CSF activates several important functions of granulocyte cells, however it does not induce L-selectin shedding.