2.4.2. Surface activation markers
Immunofluorescent staining followed by flow-cytometric anal-
ysis was used to determine markers for activation, expressed on the
membrane surface of neutrophils and monocytes, as described
previously. 6 Monocytes and neutrophils were gated based on their
CD14 and CD45 expression. Characterization of activation markers
was performed using antibodies (purchased from Beckman Coulter
(Miami, FL, USA)) directed against an adhesion molecule of the b 2
integrin family (CD11b), a degranulation marker of azurophilic
(CD63) or specific granulae (CD66b) and L-selectin (CD62L).
Immune-fluorescent staining was performed according to the “lyse
and wash” method. Flowcytometryanalyses werecarried out on an
Epics XL (Beckman Coulter, Hialeah, FL).