Cold stress restricts malaria parasite growth by affecting erythrocyte invasion

Malaria remains a major global infectious disease, and Plasmodium falciparum, the causative agent of severe falciparum malaria, is associated with high mortality in severe cases. P. falciparum is known to be highly sensitive to temperature fluctuations; however, the specific stages of the intraerythrocytic developmental cycle affected by cold stress remain poorly understood. In this study, we investigated the effects of cold stress on parasite development. Under in vitro culture conditions at 27-33 °C, parasite growth and differentiation during the intraerythrocytic stages were suppressed in a temperature-dependent manner. Experimental exposure of mice to 4 °C resulted in reduced parasitemia in a rodent malaria infection model. Cold stress enhanced growth suppression in combination with low-dose artesunate both in vitro and in vivo. Parasites exposed to cold stress exhibited decreased erythrocyte invasion efficiency, accompanied by broad downregulation of genes associated with erythrocyte invasion. Notably, exposing parasites to 27-33 °C reduced ama1 expression at both the schizont and ring stages. These findings suggest that cold stress suppresses malaria parasite growth by impairing erythrocyte invasion, highlighting temperature as a critical environmental factor regulating parasite development in warm-blooded hosts.