FP578FLOWCYTOMETRIC ANALYSIS OF MESOTHELIAL CELLS AND MACROPHAGES IN PERITONEAL DIALYSIS EFFLUENT

Le résumé fourni par la source

Introduction and Aims: Cell components of peritoneal dialysis (PD) effluent are derived from peritoneal tissue and affected by pathophysiological condition of PD peritoneum. The purpose of this study is to establish a diagnostic technique to assess the peritoneal pathophysiology by detection of phenotypic alterations of cell components in PD effluents, focusing on mesothelial cells and macrophages and their expression of podoplanin (pod), CD14 (LPS receptor) and CD44 (hyaluronan receptor). Podoplanin and CD44 are cofactors which mediate the interaction between extracellular matrices and intracellular signaling and regulate various cellular functions and responses, such as proliferation, differentiation and migration. Methods: PD effluents obtained from 3 patients initiated PD within 10 days and 3 patients treated with maintenance PD for years (5y8m, 5y9m and 12y5m). The cell fractions were collected by centrifugation of PD effluent with the volume of 300-1600ml. After lysis of red blood cells, the numbers of nucleated cells (NCC) were counted by automated cell counter (ScepterTM, Millipore). The cells were stained with the labeled antibodies for cell surface molecules, such as pod, CD14 and CD44 and analyzed by flowcytometry (FACSCalibur™, BD Biosciences). The mesothelial cell fraction was gated by positivity of pod and the macrophage fraction was gated by positivity of CD14. The expression rate (%) and fluorescence intensity (FI) of pod, CD14 and CD44 in both fractions were analyzed between induction and maintenance phases and the association with clinical findings of the patients was investigated.

Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.