Changes in RhD and CD47 expression during red blood cell aging

LUJÁN BRAJOVICH, MELINA ELIANA; TRUCCO BOGGIONE, CAROLINA; MATTALONI, STELLA MARIS; ENSINCK, MARÍA ALEJANDRA; RACCA, LILIANA; BIONDI, CLAUDIA; COTORRUELO, CARLOS

Mar del Plata

Congreso; LIX Reunión Científica Anual de la Sociedad Argentina de Investigación Clínica (SAIC). LXII Reunión Anual de la Sociedad Argentina de Inmunología (SAI); 2014

SAIC-SAI

Red blood cells (RBCs) undergo multiple changes while they age in vivo. Loss or reorganization of several proteins occurs gradually in erythrocytes membrane over 120 days in circulation. The aim of this work was to evaluate the participation of the RhD and CD47 polipeptides during the RBC senescence process. D positive samples expressing the C, c, E and e Rh antigens (n30) and controls (D, C and E negative) were incubated with three different IgM anti-D mAbs. Subsequently, anti-µ-FITC and anti-CD47-PE were added. All samples were analyzed by flow cytometry (FACSAria II). RBCs were gated on the basis of their forward scatter (FSC) and side scatter (SSC) characteristics with the use of logarithmic amplification. Flow cytometric analysis was carried out using the FACSDiva software. Dot-plot based on the FSC (cell size) versus SSC (cell density) parameters was used to identify Young (Y) and Senescent (Se) RBCs subpopulations. Statistical analysis showed that the median of the fluorescence intensity (MFI) (expressed as mean and standard deviation) associated with the D antigen expression obtained with the 3 different mAbs for YRBCs was 10635.6±917.0, 7527.6±625.5 and 9629.2±625.5. The data obtained for SeRBCs were 8075.7±1033.9, 6119.7±598.3 and 8027.3±928.1 respectively. The MFI values were significantly higher in YRBCs when compared with SeRBC (p