Granulocyte- and monocyte-platelet adhesion index in coronary and peripheral blood after extracorporeal circulation and reperfusion

Abstract

Background: Neutrophil-granulocyte and mononuclear-cell functional changes occur during cardiopulmonary bypass and cardiovascular surgery. In particular, leukocyte-platelet interaction, leading to generation of heterotypic coaggregates, represents an amplification mechanism of the local inflammatory response and tissue damage. Methods: Samples of 20 patients were drawn from venous coronary sinus before cardioplegic arrest and immediately after reperfusion, as well as from peripheral blood at 5 and 24 h postoperatively. The granulocyte and monocyte surface expression of CD162, CD15s, CD18, and CD11b were quantified by flow cytometry at the different times. Parallel variations of circulating leukocyte-platelet conjugates (percentages) and a derived (cell number-normalized) leukocyte-platelet adhesion index were measured using a combination of antibodies against CD45, CD14, and CD41a. The evaluation of platelet functional state was carried out using antibodies against CD62P (P-selectin) and PAC-1. Results: Monocyte and granulocyte cell number increased markedly in coronary blood at reperfusion and in peripheral blood postoperatively when compared with measurements done before cardioplegia. A very different course characterized the changes of the leukocyte-platelet adhesion index with respect to the variations of circulating leukocyte-platelet coaggregates (percentages). Leukocyte molecules expression showed no significant variations for CD15s on both the leukocyte subsets, while a significant up-modulation for CD162 was observed on monocytes at 24 h after extracorporeal circulation (P = 0.0002), and for CD11b on granulocytes at 5 h postoperatively (F = 0.033). A loss of CD162 expression was observed in coronary blood at reperfusion (P = 0.0038) on granulocytes, associated to a down-modulation of CD18 (P = 0.0033) and CD11b (P = 0.0184) in peripheral blood at 24 h postoperatively. No significant up-regulation of platelet activatory molecules expression was found at coronary reperfusion, as well as postoperatively in the peripheral blood, when compared with the before-cardioplegia derived data. Conclusions: The over time variations of a normalized leukocyte-platelet adhesion index seem to reflect the cumulative leukocyte-platelet functional interaction more accurately than the parallel measurements of cellular conjugates. The absence of platelet activation suggests that the leukocyte membrane modifications play a main role in controlling the formation and stability of heterotypic leukocyte-platelet coaggregates after cardiac surgery with extracorporeal circulation. © 2006 International Society for Analytical Cytology.