Plasmodium falciparum parasites exit the infected erythrocyte after haemolysis with saponin and streptolysin O
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PROTOZOOLOGY - SHORT COMMUNICATION
Plasmodium falciparum parasites exit the infected erythrocyte after haemolysis with saponin and streptolysin O Katharina A. Quadt 1,2 & Xanthoula Smyrnakou 1,2 & Friedrich Frischknecht 3,4
&
Guido Böse 1 & Markus Ganter 1,2
Received: 25 June 2020 / Accepted: 11 October 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Malaria is caused by unicellular parasites of the genus Plasmodium, which reside in erythrocytes during the clinically relevant stage of infection. To separate parasite from host cell material, haemolytic agents such as saponin are widely used. Previous electron microscopy studies on saponin-treated parasites reported both, parasites enclosed by the erythrocyte membrane and liberated from the host cell. These ambiguous reports prompted us to investigate haemolysis by live-cell time-lapse microscopy. Using either saponin or streptolysin O to lyse Plasmodium falciparum–infected erythrocytes, we found that ring-stage parasites efficiently exit the erythrocyte upon haemolysis. For late-stage parasites, we found that only approximately half were freed, supporting the previous electron microscopy studies. Immunofluorescence imaging indicated that freed parasites were surrounded by the parasitophorous vacuolar membrane. These results may be of interest for future work using haemolytic agents to enrich for parasite material. Keywords Plasmodium falciparum . Haemolysis . Saponin . Streptolysin O
Introduction Malaria is caused by unicellular parasites of the genus Plasmodium. During the clinically relevant blood stage of infection, Plasmodium spp., reside inside erythrocytes. During invasion of the erythrocyte, the parasite forms an
Section Editor: Guido Favia Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00436-020-06932-9) contains supplementary material, which is available to authorized users. * Guido Böse [email protected] * Markus Ganter [email protected] 1
Zendia GmbH, Rummler 5, 48324 Sendenhorst, Germany
2
Parasitology, Centre for Infectious Diseases, Heidelberg University Hospital, Im Neuenheimer Feld 324, 69120 Heidelberg, Germany
3
Integrative Parasitology, Centre for Infectious Diseases, Heidelberg University Hospital, Im Neuenheimer Feld 324, 69120 Heidelberg, Germany
4
German Centre for Infection Research, Heidelberg Division, Heidelberg, Germany
intracellular compartment, the parasitophorous vacuole (PV), which is delimitated by the parasitophorous vacuolar membrane (PVM) (Matz et al. 2020). These intracellular stages of Plasmodium pose a challenge for assays, which are sensitive to contaminations with host cell material, such as experimental immunisations as well as proteomic and biochemical approaches. Different attempts have been made to separate the parasite from host cell material, including hypotonic lysis, ammonium chloride lysis and mechanical lysis with French pressure cell (Cook et al. 1969; Martin et al. 1971). A very widely used substance to
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