Inhibition of platelet phagocytosis as an in vitro predictor for therapeutic potential of RBC antibodies in murine ITP

Abstract

Polyclonal anti-D is a first-line therapy for immune thrombocytopenia (ITP). Monoclonal antibodies are desirable alternatives, but none have yet proven successful despite their ability to opsonize erythrocytes (or red blood cells, RBCs) and cause anemia. Here, we examined 12 murine erythrocyte–specific antibodies of different specificity and subtypes and found that 8 of these antibodies could induce anemia in antigen-positive mice. Of these 8 antibodies, only 5 ameliorated ITP. All antibodies were examined for their in vitro ability to support macrophage-mediated phagocytosis of erythrocytes. Antibodies which supported erythrocyte phagocytosis in vitro successfully ameliorated ITP in vivo. To examine the ability of each antibody to inhibit phagocytosis of platelets, the antibodies were used to sensitize erythrocytes in vitro and these were added to a platelet phagocytosis assay. Antibodies that inhibited platelet phagocytosis in vitro also all ameliorated ITP in vivo. We conclude that inducing anemia is not a sufficient condition for amelioration of ITP but that the antibody’s ability to prevent platelet phagocytosis in vitro predicted its ability to ameliorate ITP. We suggest that inhibition of in vitro platelet phagocytosis may prove to be a valuable tool for determining which erythrocyte antibodies would likely be candidates for clinical use in ITP.

Human Fc gamma receptor IIIA blockade inhibits platelet destruction in a humanized murine model of ITP

“…Various therapeutic approaches have been explored to target FcγRs. IV immunoglobulin and anti-D are both FDA-approved treatments for ITP and can be considered to exert their effects via FcγR blockade, 11 , 21 although the precise mechanism of both treatments is not fully understood.…”

“…Antibodies 3G8, 10.1, AT10, IV.3, 17C02-IgG2a, and isotypes controls (IgG1 and IgG2a) at 0.5 mg/mL in phosphate-buffered saline (PBS; Gibco) were fully Fc region deglycosylated as previously described. 11 …”