Potential impact of complement regulator deficiencies on hemolytic reactions due to minor ABO-mismatched transfusions

Abstract

AbstractMinor ABO-mismatched transfusions are a common occurrence, although infrequent transfusion reactions occur. We sought to investigate the regulation of complement C3 activation induced by anti-A. In vitro complement C3 activation was observed with 10 of 30 group O samples and correlated with immunoglobulin M (IgM) anti-A titers. We developed an in vitro paroxysmal nocturnal hemoglobinuria (PNH) model of hemolysis in which group A1 red blood cells (RBCs) were chemically treated with 2-aminoethylisothiouronium (AET) to alter regulators of complement C3 activation. Intravascular hemolysis was simulated by incubating an IgG nonhemolytic group O plasma (titer = 32) with A1 RBCs. IgG was detected on the RBCs, but hemolysis was observed with AET-treated RBCs only. When treated and untreated RBCs were tested together (1:4), we determined that the failure to observe C3b/d deposition on RBCs was due to the complete hemolysis of the AET-treated minor RBC population. A group O patient with a 9% CD59-deficient PNH clone was sensitized with an IgM anti-I. Hemolysis, with a weak positive direct antiglobulin test (DAT) resulting from C3b/d, was observed after incubation with fresh AB serum. Flow cytometry showed an 86% reduction of the PNH clone. Our work indicates that the transfusion of minor ABO-mismatched plasma could cause hemolysis with a negative DAT C3b/d. We propose that the presence of a PNH clone is 1 possible cause of unexplained anemia for recipients of ABO-mismatched product. This work suggests that other acquired or inherited defects of decay-accelerating factor and membrane inhibitor of reactive lysis could be responsible for infrequent but clinically important hemolysis after ABO-mismatched transfusions.