An Opsonic Phagocytosis Assay for Plasmodium falciparum Sporozoites

Plasmodium falciparum malaria remains the deadliest parasitic disease worldwide. Vaccines targeting the preerythrocytic sporozoite and liver stages have the potential to entirely prevent blood-stage infection and disease, as well as onward transmission. Sporozoite surface and secreted proteins are leading candidates for inclusion in a preerythrocytic stage-specific, antibody-based vaccine. Preclinical functional assays to identify humoral correlates of protectionin vitro and to validate novel sporozoite protein targets for inclusion in multisubunit vaccines currently do not consider the interaction of sporozoite-targeting antibodies with other components of the immune system. Here, we describe the development of a simple flow cytometric assay to quantitatively assess the ability of antibodies directed againstP. falciparum sporozoites to facilitate their phagocytosis. We demonstrate that thiss porozoiteo psonicp hagocytosisa ssay (SOPA) is compatible with both monoclonal antibodies and human immune serum and can be performed using cryopreservedP. falciparum sporozoites. This simple, accessible assay will aid with the assessment of antibody responses to vaccination withPlasmodium antigens and their interaction with phagocytic cells of the immune system.