During the procedures of centrifugation leukapheresis and plateletpheresis, donors occasionally experience adverse clinical reactions. The possibility of whether the activation of granulocytes and the subsequent release reactions, which may have been triggered by this extracorporeal circuit, were responsible for these adverse effects was evaluated. Six blood samples were obtained at set intervals during cytapheresis. Of these samples, four were taken directly from the donor. The remaining two were drawn from the efferent lines, i.e., those which return blood from the cytapheresis machine to the donor. Reactive oxygen species produced by granulocytes were measured by chemiluminescence (CL) using microamounts of whole blood or isolated granulocytes. Furthermore, secreted granulocyte products such as neutral proteinase elastase, which is present in plasma in a complex with alpha-1-proteinase inhibitor (E-alpha-1-PI), and lysosomal beta-glucuronidase were examined. A complete blood cell count and the values of hemoglobin, hematocrit, lactate dehydrogenase, protein, albumin, and proteinase inhibitors such as alpha-2-macroglobulin and alpha-1-proteinase inhibitor were also determined. Clinical chemical and cytologic values, with the exception of those for E-alpha-1-PI, were 10 to 17 percent lower than values before apheresis. These results can be attributed to inherent plasma volume expansion. Reduced CL was observed on the stimulation of phagocytes in the whole blood assay, as well as with stimulated granulocytes. Unstimulated granulocytes, on the other hand, showed an increased native CL. These data do not indicate a cytapheresis-mediated activation of the oxidative metabolism of granulocytes, and the concomitant discharge of proteolytic enzymes remains, therefore, of no clinical importance.

Summary:During the procedure of centrifugation cytapheresis donors occasionally experience adverse clincial reactions. We evaluated the possibility of whether activation of granulocytes and their subsequent release reactions, which may have been triggered by this extracorporeal circuit, were responsible for these adverse effects. Six blood samples were obtained during various set intervals during plateletapheresis. Of these, four samples were taken directly from each donor. The remaining two were drawn from the efferent lines, i.e. those which return blood from the cytapheresis machine back to the donor. Reactive oxygen species produced by granulocytes were monitored by chemiluminescence using microamounts of whole blood or isolated granulocytes. Furthermore, secreted granulocyte products such as neutral proteinase elastase, present in plasma in a complex with oti-proteinase inhibitor (complexed elastase), and lysosomal -glucuronidase were examined. A complete blood cell count, as well as values of haemoglobin, haematocrit, lactate dehydrogenase, protein, albumin and proteinase inhibitors such as oc 2 -macroglobulin and αϊ-proteinase inhibitor were also determined. Complexed elastase increased from a preapheresis value of about 140 μg/l to about 180 μg/l at the end of the cytapheresis. All other clinical chemical and cytological values were 8 to 12 percent lower than preapheresis values, which can be attributed to inherent plasma volume expansion. Reduced chemiluminescence was observed upon stimulation of phagocytes in the whole blood assay (about 700 counts/min χ ΙΟ 3 χ 50000 cells vs. about 600 counts/min χ ΙΟ 3 χ 50000 cells). This decrease was also seen with stimulated granulocytes (about 5800 counts/min χ ΙΟ 3 χ 50000 cells vs. about 4500 counts/min χ ΙΟ 3 χ 50000 cells). Unstimulated granulocytes, on the other hand, showed an increased native chemiluminescence. These data do not, however, indicate a cytapheresis mediated activation of the oxidative metabolism of granulocytes. Concomitant discharge of proteolytic enzymes remains, therefore, of no clinical importance.