Uptake of C5a by polymorphonuclear leukocytes (PMNs) after focal cerebral ischemia. I. Effect of tirilazad mesylate intervention on C5a uptake by PMNs.

PMNs are believed to play an important role in ischemia and C5a uptake is an important functional indicator for G protein-coupled receptor trafficking. The ability of PMNs to internalize 125I-labeled C5a in vitro is an index of the functional state of these cells. We evaluated the effects of model preparation and focal cerebral ischemia by middle cerebral artery occlusion and reperfusion (MCA:O/R) on internalization of C5a by PMNs isolated from baboons (Papio anubis/cynocephalus). Similar assays were performed on PMNs isolated before and after exposing the animals undergoing MCA:O/R to an anti-inflammatory 21-aminosteroid, tirilazad mesylate (U74006F). Surgical implantation of the MCA occlusion device had no measureable effect on uptake of C5a to the cytosol by the PMN. In contrast, MCA:O/R appeared to decrease uptake of C5a. Both in vivo and in vitro administration of tirilazad, to otherwise untreated animals and to isolated cells, respectively, reduced baseline values of C5a uptake in the PMNs. Cytosolic uptake of C5a was also reduced in PMNs isolated from subjects that had undergone MCA:O/R and tirilazad treatment. These results suggest that focal cerebral ischemia, with or without exposure to tirilazad mesylate, may inhibit internalization of C5a by the PMN receptors. The effects of stroke on the ability of C5a to gain entry into the PMN may result from receptor down-regulation or "desensitization" of the cell, possibly due to activation of complement and generation of C5a which occupied the receptors. Alternatively, the effect of tirilazad presumably results from the ability of this drug to enter the membrane lipid layer and reduce fluidity.