Activation of human polymorphonuclear cells induces formation of functional gap junctions and expression of connexins.

摘要: BACKGROUND During inflammatory responses activated polymorphonuclear cells (PMNs) adhere to each other and form clusters within the vasculature or injured tissues. We hypothesized that conditions partially mimic chemical environment of foci induce expression functional gap junctions (GJs) between cultured PMNs. MATERIAL/METHODS Human PMNs were treated with bacterial lipopolysaccharide (LPS), TNF-a, LPS plus medium conditioned by LPS-treated endothelial (ECs) TNF-a ECs medium. Gap junctional communication was evaluated dye coupling technique using a permeant an impermeant GJ fluorescent blockers. The connexins, protein subunits, immunocytochemistry immunoblotting. Cytochalasin-D nocodazole used evaluate involvement cytoskeleton in induction coupling. RESULTS Treatment induced formation PMN aggregates, but not coupled. If latter protocols occurred resting ECs, respectively, intercellular transfer only molecule observed most clustered cells. Dye reversibly inhibited blockers prevented cytochalasin-D, microfilament disrupter, nocodazole, microtubule disrupter. Treatments also connexin43 connexin40, connexin32 immunoreactivity. None these connexins detected circulating CONCLUSIONS EC-derived factor(s) integrity are required for LPS- TNF-a-treated is correlated presence 43 40, 32 requires intact microfilaments.