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J Med Microbiol 56 (2007), 629-636; DOI: 10.1099/jmm.0.47039-0
© 2007 Society for General Microbiology
ISSN 1473-5644

Aloe vera leaf exudate induces a caspase-independent cell death in Leishmania donovani promastigotes

Avijit Dutta1,{dagger}, Suman Bandyopadhyay1,{ddagger}, Chitra Mandal1 and Mitali Chatterjee2

1 Department of Immunobiology, Indian Institute of Chemical Biology, 4 Raja SC Mullick Road, Kolkata 700 032, India

2 Department of Pharmacology, Institute of Postgraduate Medical Education & Research, 244 B Acharya JC Bose Road, Kolkata 700 020, India

Correspondence
Chitra Mandal
cmandal{at}iicb.res.in
Mitali Chatterjee
ilatim{at}vsnl.net

Received 30 October 2006
Accepted 12 December 2006

Leishmaniasis constitutes a complex of diseases with clinical and epidemiological diversity and includes visceral leishmaniasis, a disease that is fatal when left untreated. In earlier studies, the authors reported that Aloe vera leaf exudate (AVL) is a potent antileishmanial agent effective in promastigotes of Leishmania braziliensis, Leishmania mexicana, Leishmania tropica, Leishmania major and Leishmania infantum and also in axenic amastigotes of Leishmania donovani. In the present study, it has been demonstrated that, in promastigotes of L. donovani (IC50=110 µg ml–1), AVL mediates this leishmanicidal effect by triggering a programmed cell death. Incubation of promastigotes with AVL caused translocation of phosphatidylserine to the outer leaflet of the plasma membrane as measured by annexin V binding, which was accompanied by loss of mitochondrial membrane potential, release of cytochrome c into the cytosol and concomitant nuclear alterations that included chromatin condensation, deoxynucleotidyltransferase-mediated dUTP end labelling and DNA laddering. As this AVL-induced leishmanicidal effect could not be inhibited by protease inhibitors including Z-Val-Ala-DL-Asp (methoxy)-fluoromethylketone, a broad-spectrum caspase inhibitor, non-involvement of caspases and major proteases was suggested. Additionally, AVL treatment caused no increase in cytosolic Ca2+ or generation of reactive oxygen species, indicating that although promastigote death was induced by an apoptotic-like mechanism similar to metazoan apoptosis, the pathways of induction and/or execution differed at the molecular level.

Abbreviations: AVL, Aloe vera leaf exudate; H2DCFDA, 2',7'-dichlorodihydrofluorescein diacetate; MFI, mean fluorescence intensity; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; PCD, programmed cell death; PI, propidium iodide; ROS, reactive oxygen species; TUNEL, terminal deoxynucleotidyltransferase mediated dUTP end labelling.

{dagger}Present address: Dept of Pharmacology, Institute of Postgraduate Medical Education & Research, 244 B Acharya JC Bose Road, Kolkata 700 020, India.

{ddagger}Present address: Center for Immunotherapy of Cancer and Infectious Diseases, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030-1601, USA.




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