CD47 expression on human erythrocytes of different ages

ENSINCK A; LUJÁN BRAJOVICH M; GARCÍA BORRÁS S; COTORRUELO C; BIONDI C

Rosario

Congreso; XX Congreso y XXXVIII Reunión Anual de la Sociedad de Biología de Rosario; 2018

Sociedad de Biología de Rosario

CD47 expression on red blood cells (RBCs) is an antiphagocytic signal for the macrophages. The senescence process of these cellsinvolves a series of biochemical changes leading to greater phagocytosis susceptibility. The CD47 diminished expression during RBCsaging would be an alternative mechanism for clearance of RBCS. The aim of this study was to evaluate the expression levels of CD47 by flow cytometry (FCM) on RBCs population from different ages. CD47 mean intensity fluorescence (MIF) analysis was performed by FCM based on the FSC (cell size) versus SSC (cell density) parameters to study RBCs populations of different sizes and density.Given that FSC represents cell size and SSC internal cellular complexity, we assume that the population of senescent RBC (SeRBC) is located in the < FSC and > SSC and the young RBC (YRBC) population in >FSC and < SSC. ACD anticoagulated blood samples from normal volunteer donors (n=30). Flow cytometric analyses were performed on a FACSAria II flow cytometer using the FACSDiva software for acquisition and analysis. 50 μL of 0.2% PBS RBCs suspension were incubated with 5 μL of anti-human CD47 PE in darkness at room temperature for 30 min. After staining, cells were washed twice and resuspended in 0.5 mL PBS and analyzed byflow cytometry. The CD47 MIFs (standard deviation) were significantly lower for SeRBC 11010.0 (882.9) compared with YRBC 11965.0 (945.7) (p