Antibacterial activity of erythrocyte from grass carp (Ctenopharyngodon idella) is associated with phagocytosis and reactive oxygen species generation

Red blood cells (RBCs) are widely accepted as their primary function in respiration. Recent studies in mammals have revealed a vital role in immune responses of ABCs; however, little is known about immune function of teleost erythrocytes. Here we demonstrated that RBCs from grass carp (Ctenopharyngodon idella) were capable of binding and aggregating the bacteria with apparent morphological alterations. The phagocytosis by teleost RBCs (erythrophagocytosis) was visualized by confocal, scanning and transmission electron microscopy. Hb-Fe-II of hemoglobin (Hb) could quickly be auto-oxidated to Hb-Fe-III (methemoglobin/metHb) in the presence of oxygen (O-2), and release superoxide radical (O-2(-)) which could be spontaneously dismutated into H2O2 that could further oxidize Hb-Feln to transient HbFe(IV)-OH (ferryl-Hb). Furthermore, bacterial extracellular proteases and pathogen-associated molecular patterns (PAMPs) binding to Hb could synergistically activate pseudoperoxidase, subsequently facilitated the generation of reactive oxygen species (ROS) which were toxic to the bacteria. Our results indicated that erythrocyte pertains anti-bacterial activity using unique ROS generation pathway via oxidation of hemoglobin and associated with its phagocytosis.