2ND MESSENGER FORMATION ALTERED BY DIFFERENT DIALYSIS MEMBRANES IN HUMAN-LEUKOCYTES

Second messenger formation altered by different dialysis membranes in human leukocytes. A number of agents stimulate transmembrane cell signaling in different cell types through the formation of the second messenger diacylglycerol (DAG) which activates protein kinase C (PKC). The aim of this study was to investigate phospholipase C activation, DAG formation, and cellular adhesion to dialysis membranes after simulated dialytic treatment of either human leukocytes or clonal hematopoietic cells. Cells were circulated for 60 minutes in a closed-loop dialysis system using three different dialyzers: cuprophan (CU), polysulphone (PS), and AN69 (PAN). Another cell aliquot was left within the dialyzers without circulation. Samples were taken at different time intervals and cells counted. Cells were labeled with tritiated glycerol overnight, and DAG was measured by thin-layer chromatography. Our data showed that cells tended to adhere with more efficiency to CU than to the synthetic dialyzers. Circulation in the in vitro dialysis circuit resulted in the rapid (5 min) formation of [H-3]DAG (CU 1.95-; PS 1.34-; PAN 1.24-fold increase over untreated cells). The DAG level peaked at 15 to 30 minutes and remained constant thereafter (CU 1.70; PS 1.96; PAN 1.66). When we measured DAG formation in cells that had been kept in the dialyzers without circulation, we found that cells exposed to CU showed a much higher and rapid activation than those exposed to PS or PAN, as if CU per se was able to activate early cell signaling (CU 1.95-; PS 0.97-; PAN 1.09-; DAG, -fold increase over control). These data indicate that exposure to, and circulation through dialyzers activated early transmembrane signaling in human leukocytes, and that different dialysis membranes showed distinct effects on cell adhesion and stimulation.