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Open Access Research article

Cytotoxicity of purified listeriolysin O on mouse and human leukocytes and leukaemia cells

Radosław Stachowiak1, Marcin Łyżniak2, Maja Grabowska1, Katarzyna Roeske1, Tomasz Jagielski1, Jacek Bielecki1, Bożena K Budziszewska3, Grażyna Hoser2 and Jerzy Kawiak2*

Author Affiliations

1 Department of Applied Microbiology, Institute of Microbiology, Faculty of Biology, University of Warsaw, I. Miecznikowa 1, 02-096 Warsaw, Poland

2 Medical Centre of Postgraduate Education, Marymoncka 99/103, 01-813 Warsaw, Poland

3 Department of Hematology, Institute of Hematology and Transfusion Medicine, I. Gandhi 14, 02-776 Warsaw, Poland

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BMC Biotechnology 2014, 14:77  doi:10.1186/1472-6750-14-77

Published: 18 August 2014



Listeriolysin O (LLO) is the main virulence factor of Listeria monocytogenes and facilitates the intracellular survival of the pathogen. Some of its characteristics endorse the growing popularity of LLO for use in biotechnology, particularly in the development of novel vaccines. Here, we evaluate the use of LLO to eradicate leukaemia cells.


A purified LLO preparation was obtained by affinity chromatography. The LLO preparation procedure was optimized and purified LLO was tested for optimal conditions of storage including temperature, application of proteinase inhibitors and serum components. We demonstrated the possibility of regulating LLO activity by adjusting cell membrane cholesterol content. The LLO preparation had haemolytic activity and had a cytotoxic effect on the human T-leukaemia Jurkat cell line as well as mouse and human peripheral blood mononuclear cells.


LLO has a very potent cytotoxic activity towards human leukocytes. Importantly, the cytotoxic activity was easily regulated in vitro and could be restricted to areas containing malignant cells, raising the possibility of future clinical application of LLO for leukaemia treatment.

Listeriolysin O; Protein purification; Cytotoxicity; Leukocytes; Leukaemia