Antibody responses to Plasmodium vivax Duffy binding and Erythrocyte binding proteins predict risk of infection and are associated with protection from clinical Malaria

Background The Plasmodium vivax Duffy Binding Protein (PvDBP) is a key target of naturally acquired immunity. However, region II of PvDBP, which contains the receptor-binding site, is highly polymorphic. The natural acquisition of antibodies to different variants of PvDBP region II (PvDBPII), including the AH, O, P and Sal1 alleles, the central region III-V (PvDBPIII-V), and P. vivax Erythrocyte Binding Protein region II (PvEBPII) and their associations with risk of clinical P. vivax malaria are not well understood. Methodology Total IgG and IgG subclasses 1, 2, and 3 that recognize four alleles of PvDBPII (AH, O, P, and Sal1), PvDBPIII-V and PvEBPII were measured in samples collected from a cohort of 1 to 3 year old Papua New Guinean (PNG) children living in a highly endemic area of PNG. The levels of binding inhibitory antibodies (BIAbs) to PvDBPII (AH, O, and Sal1) were also tested in a subset of children. The association of presence of IgG with age, cumulative exposure (measured as the product of age and malaria infections during follow-up) and prospective risk of clinical malaria were evaluated. Results The increase in antigen-specific total IgG, IgG1, and IgG3 with age and cumulative exposure was only observed for PvDBPII AH and PvEBPII. High levels of total IgG and predominant subclass IgG3 specific for PvDBPII AH were associated with decreased incidence of clinical P. vivax episodes (aIRR = 0.56-0.68, P0.001-0.021). High levels of total IgG and IgG1 to PvEBPII correlated strongly with protection against clinical vivax malaria compared with IgGs against all PvDBPII variants (aIRR = 0.38, P<0.001). Antibodies to PvDBPII AH and PvEBPII showed evidence of an additive effect, with a joint protective association of 70%. Conclusion Antibodies to the key parasite invasion ligands PvDBPII and PvEBPII are good correlates of protection against P. vivax malaria in PNG. This further strengthens the rationale for inclusion of PvDBPII in a recombinant subunit vaccine for P. vivax malaria and highlights the need for further functional studies to determine the potential of PvEBPII as a component of a subunit vaccine for P. vivax malaria. Author summary Plasmodium vivax is responsible for most malaria infections outside Africa, with 13.8 million vivax malaria cases reported annually worldwide. Antibodies are a key component of the host response to P. vivax infection, and their study can assist in identifying suitable vaccine candidates and serological biomarkers for malaria surveillance. The binding of P. vivax Duffy binding protein region II (PvDBPII) to the Duffy Antigen Receptor for Chemokines (DARC) is critical for P. vivax invasion of reticulocytes. Although the binding residues for DARC are highly conserved across PvDBPII, the parasite displays high sequence diversity in non-binding residues of PvDBPII. Other regions such as PvDBPIII-V are relatively conserved. Recently, sequencing of P. vivax field isolates, identified a homologous erythrocyte-binding protein (PvEBP), which harbors a domain, region II (PvEBPII), that is homologous to PvDBPII. To date, there has been limited investigation into the naturally acquired immunity to both PvDBPIII-V and PvEBPII in human populations. Using a longitudinal cohort study, we have characterized the serological response to PvDBPII, PvDBPIII-V, and PvEBPII among 1-3 years old PNG children and investigated associations with protection against clinical malaria. This study shows that both total IgG and IgG3 to the predominant PvDBPII AH allele in PNG, and total IgG and IgG1 to PvEBPII were associated with protection from P. vivax malaria.