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Background  Vivax malaria is associated with significant morbidity and economic loss, and constitutes the bulk of malaria cases in large parts of Asia and South America as well as recent case reports in Africa. The widespread prevalence of vivax is a challenge to global malaria elimination programmes. Vivax malaria control is particularly challenged by existence of dormant liver stage forms that are difficult to treat and are responsible for multiple relapses, growing drug resistance to the asexual blood stages and host-genetic factors that preclude use of specific drugs like primaquine capable of targeting  Plasmodium vivax  liver stages. Despite an obligatory liver-stage in the  Plasmodium  life cycle, both the difficulty in obtaining  P. vivax  sporozoites and the limited availability of robust host cell models permissive to  P. vivax  infection are responsible for the limited knowledge of hypnozoite formation biology and relapse mechanisms, as well as the limited capability to do drug screening. Although India accounts for about half of vivax malaria cases world-wide, very little is known about the vivax liver stage forms in the context of Indian clinical isolates.    Methods  To address this, methods were established to obtain infective  P. vivax  sporozoites from an endemic region in India and multiple assay platforms set up to detect and characterize vivax liver stage forms. Different hepatoma cell lines, including the widely used HCO4 cells, primary human hepatocytes as well as hepatocytes obtained from iPSC’s generated from vivax patients and healthy donors were tested for infectivity with  P. vivax  sporozoites.    Results  Both large and small forms of vivax liver stage are detected in these assays, although the infectivity obtained in these platforms are low.    Conclusions  This study provides a proof of concept for detecting liver stage  P. vivax  and provide the first characterization of  P. vivax  liver stage forms from an endemic region in India.

Plasmodium vivax liver stage assay platforms using Indian clinical isolates