Population genetic structure and polymorphism analysis of gene encoding apical membrane antigen-1 (AMA-1) of Iranian Plasmodium vivax wild isolates.

Plasmodium vivax apical membrane antigen-1 (PvAMA-1) is a major candidate antigen for human malaria vaccine. In the present study, polymorphism of pvama-1 among Iranian isolates was investigated to generate useful information on this vaccine candidate antigen, which is required for the rational design of a vaccine against P. vivax. Blood samples were collected from P. vivax-infected Iranian patients during 2009-2010. Of 99 collected isolates, 37 were analyzed for almost the entire pvama-1 gene using sequencing. The overall nucleotide diversity (π) was 0.00826 ± 0.0004 and the majority of polymorphic sites were identified in domain I (DI) of the pvama-1 gene. Neutrality analysis using Tajima's D, Fu and Li's D* and F* and McDonald Kreitman tests showed a significant positive departure from neutral substitution patterns, indicating a possible balancing selection across the entire ectodomain and DI sequences of pvama-1 gene. However, no evidence was found for the balancing selection in DII and DIII regions of Iranian PvAMA-1. Also, 29 haplotypes with different frequencies were identified and the overall haplotype diversity was 0.982 ± 0.012. Epitope mapping prediction of PvAMA-1 showed the potential B-cell epitopes across DI-DIII overlap with E145K, P210S, R249H, G253E, K352E, R438H and N445D mutations; however, no mutation has been found in intrinsically unstructured/disordered regions. The fixation index (Fst) estimation between Iran and the closest geographical sites such as India (0.0707) showed a slight geographical genetic differentiation; however, the Fst estimation between Iran and Thailand (0.1253) suggested a moderate geographical isolation. In summary, genetic investigation in pvama-1 among Iranian P. vivax isolates indicates that this antigen showed limited antigenic diversity and most of the detected mutations are located outside B-cell epitopes. Therefore, the present results have significant implications in understanding the nature of P. vivax population circulating in Iran as well as in providing useful information for malaria vaccine development based on this antigen.