A monoclonal antibody to rhesus erythrocyte band 3 inhibits invasion by malaria (Plasmodium knowlesi) merozoites

Abstract

Receptors on erythrocytes and malaria parasites mediate specific attachment and junction formation between these cells that lead to invasion of the erythrocytes. We identified monoclonal antibody A9 and its subclone A9D3 that bound to rhesus erythrocytes and blocked invasion of the erythrocytes by Plasmodium knowlesi merozoites. The monoclonal antibodies did not block attachment, the initial step in invasion, although swelling and crenation of the erythrocyte, which normally occur after attachment, were rarely observed in the presence of antibody. The monoclonal antibody immunoprecipitated rhesus erythrocyte band 3. It bound to erythrocytes of another Old World monkey, the kra monkey, but not to erythrocytes of New World monkeys, chimpanzees, or man. Since the antibody did not bind to human erythrocytes, we could test for nonspecific toxicity to the parasite by studying the effect of the ascites and purified antibody on invasion of human erythrocytes. The antibody caused a minimal reduction in invasion of human erythrocytes, a reduction no greater than that seen with an unrelated monoclonal antibody. Further evidence that the inhibition was specific came from study of Fab fragments of A9D3. Column-purified Fab fragments reduced invasion of rhesus erythrocytes without affecting invasion of human erythrocytes. Fab fragments preabsorbed with rhesus erythrocytes did not inhibit invasion. From the above data, we conclude that band 3 is involved in a stage in the invasion process after initial recognition.