First continuous in vitro culture of Plasmodium coatneyi reveals insights into barriers to human infection

Line graph shows changes in Plasmodium coatneyi parasitemia over 40 days in macaque RBCs during in vitro culture; inset image displays RBCs with parasites indicated by red arrows.

Plasmodium coatneyi is an important model for severe malaria due to its P. falciparum-like sequestration, yet research has been limited by the lack of in vitro culture systems. Recently, molecular evidence of human P. coatneyi infections (3.5%, 3/85 of identified simian malarias in Malaysian archival samples), suggested a zoonotic potential. We report the first continuous in vitro culture of P. coatneyi and characterize its zoonotic potential using a dual-colour flow cytometry assay. Using CellTracker Deep Red-labelled target cells, we quantified invasion across primate species, revealing potential sample-specific plasticity. Robust continuous culture was established in Macaca fascicularis erythrocytes for over 40 days, with parasitaemia reaching 3% following cryopreservation recovery. While P. coatneyi successfully invaded human erythrocytes, asexual development was arrested post-invasion, with parasitaemia falling below detection limits by day 9. Analysis by microscopy confirmed a failure to complete the 48-hour developmental cycle in human cells, identifying a putative biological barrier to sustained zoonotic transmission. Ultimately, the cultivation protocol provides a sustainable in vitro platform for functional investigations, particularly severe malaria pathogenesis, and for monitoring host promiscuity in zoonotic malaria parasites.

Rossarin Suwanarusk, Adeline C Y Chua, Chukwuma S Ezenwanne, Georges Snounou, Benoît Malleret, Varakorn Kosaisavee, Eric D Lombardini, Rawiwan Imerbsin, Piyanate Sunyakumthorn, Yuko Katakai, Miako Sakaguchi, Dennis E Kyle, Satoru Kawai, Takeshi Annoura, Laurent Rénia, Pablo Bifani, Osamu Kaneko, Bruce Russell. Int J Parasitol
. 2026 May 27:104892. doi: 10.1016/j.ijpara.2026.104892.