Quantitation of surface CD14 on human monocytes and neutrophils.

摘要: The absolute number of membrane-expressed CD14, the most important endotoxin receptor, on human monocytes and neutrophils shows remarkable variation in literature. To quantify these numbers two fluorescence methods using fluorescein isothiocyanate (FITC)-labeled monoclonal antibodies (mAb) were applied. A commercially available set standard beads was used flow cytometry to quantitate CD14 with eight different mAbs. Independent from their isotype various mAbs showed minor differences indicated that peripheral blood expressed 99,500-134,600 (115,400 +/- 10,600) 1,900-4,400 (3,300 800) receptors. There no significant difference expression leukocytes unprocessed freshly obtained whole after a Ficoll isolation procedure. However, short temperature shift resulted 1.3- 1.6-fold up-regulation CD14. results reference verified Scatchard analysis spectrofluorometry mAb 26ic-FITC 109,500 per monocyte 6,700 neutrophil. For comparison monocytic THP-1 cells Fc gamma-receptors determined gave comparable published data. Our data indicate resting express roughly 110,000 molecules surface simple fluorometric assay. Correct determination other cell receptors is importance monitoring septic patients.