Severe malaria caused by Plasmodium falciparum poses a major global health 
problem with high morbidity and mortality. P. falciparum harbors a family of 
pore-forming proteins (PFPs), known as perforin like proteins (PLPs), which are 
structurally equivalent to prokaryotic PFPs. These PLPs are secreted from the 
parasites and, they contribute to disease pathogenesis by interacting with host 
cells. The severe malaria pathogenesis is associated with the dysfunction of 
various barrier cells, including endothelial cells (EC). Several factors, 
including PLPs secreted by parasites, contribute to the host cell dysfunction. 
Herein, we have tested the hypothesis that PLPs mediate dysfunction of barrier 
cells and might have a role in disease pathogenesis. We analyzed various 
dysfunctions in barrier cells following rPLP2 exposure and demonstrate that it 
causes an increase in intracellular Ca2+ levels. Additionally, rPLP2 exposed 
barrier cells displayed features of cell death, including Annexin/PI positivity, 
depolarized the mitochondrial membrane potential, and ROS generation. We have 
further performed the time-lapse video microscopy of barrier cells and found 
that the treatment of rPLP2 triggers their membrane blebbing. The cytoplasmic 
localization of HMGB1, a marker of necrosis, further confirmed the necrotic type 
of cell death. This study highlights the role of parasite factor PLP in 
endothelial dysfunction and provides a rationale for the design of adjunct 
therapies against severe malaria.