High pathogenic potential of low-affinity autoantibodies in experimental autoimmune hemolytic anemia

To assess the potency of low-affinity anti-red blood cell (RBC) autoantibod ies in the induction of anemia, we generated an immunoglobulin (Ig)G2a clas s-switch variant of a 4C8 IgM anti-mouse RBC autoantibody, and compared its pathogenic potential with that of its IgM isotype and a high-affinity 34-3 C IgG2a autoantibody. The RBC-binding activity of the 4C8 IgG2a variant was barely detectable, at least 1,000 times lower than that of its IgM isotype , having a high-binding avidity, and that of the 34-3C IgG2a monoclonal ant ibody (mAb). This low-affinity feature of the 4C8 mAb was consistent with t he lack of detection of opsonized RBCs in the circulating blood from the 4C 8 IgG2a-injected mice. However, the 4C8 IgG2a variant was highly pathogenic , as potent as its IgM isotype and the 34-3C IgG2a mAb, due to its capacity to interact with Fc receptors involved in erythrophagocytosis. In addition , our results indicated that the pentameric form of the low-affinity ISM is otype, by promoting the binding and agglutination of RBCs, is critical for its pathogenic activity. Demonstration of the remarkably high pathogenic po tency of low-affinity autoantibodies, if combined with appropriate heavy ch ain effector functions, highlights the critical role of the Ig heavy chain constant regions, but the relatively minor role of autoantigen-binding affi nities, in autoimmune hemolytic anemia.