Optimal Conditions for the Monocyte Monolayer Assay and Applications of the Assay

Immune-mediated hemolysis, whether due to transfusion of incompatible red blood cells (RBCs) in hemolytic transfusion reactions or passive transfer of anti-RBC antibodies through intravenous immunoglobulin (IVIG) therapy, contributes to complications and comorbidities in patients. A major mechanism of RBC destruction is Fc gamma receptor (FcÎłR)-dependent phagocytic removal of antibody-bound RBCs by monocyte-macrophage within the mononuclear phagocyte system. The monocyte monolayer assay (MMA) assesses FcÎłR-mediated destruction using primary monocytes from whole blood and predicts in vivo survival of serologically incompatible, allogeneic RBCs. Optimal conditions for MMA were examined in this thesis. Whole blood collected in acid citrate dextrose anticoagulant and stored at room temperature for up to 36 hours best preserved monocyte phagocytic function. Such conditions were validated using clinical samples from patients receiving IVIG. The findings suggest stored autologous patient samples yield clinically relevant MMA results, and thus permit the shipping and testing of properly stored samples.M.Sc