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Antimicrobial Agents and Chemotherapy, November 2005, p. 4474-4484, Vol. 49, No. 11
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.11.4474-4484.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Photodynamic Sensitization of Leishmania amazonensis in both Extracellular and Intracellular Stages with Aluminum Phthalocyanine Chloride for Photolysis In Vitro

Sujoy Dutta, Debalina Ray, Bala K. Kolli, and Kwang-Poo Chang*

Department of Microbiology/Immunology, Chicago Medical School, Rosalind Franklin University, 3333 Green Bay Rd., N. Chicago, Illinois 60064

Received 28 April 2005/ Returned for modification 3 June 2005/ Accepted 2 August 2005

Leishmania amazonensis, a causative agent of cutaneous leishmaniasis, is susceptible in vitro to light-mediated cytolysis in the presence of or after pretreatment with the photosensitizer aluminum phthalocyanine chloride. Cytolysis of both promastigotes and axenic amastigotes required less photosensitizer (e.g., one µg · ml–1) and a lower light dose (e.g., 1.5 J · cm–2) than did the mammalian cells examined for comparison. Exposure of Leishmania cells to the photosensitizer alone had little effect on their viability, as judged from their motility, growth, and/or retention of green fluorescent proteins genetically engineered for episomal expression. Fluorimetric assays for cell-associated and released green fluorescence proteins proved to be even more sensitive for the evaluation of cell viability than microscopy for the evaluation of motility and/or integrity. Axenic amastigotes pretreated with the photosensitizer infected macrophages of the J774 line but were lysed intracellularly when the infected cells were exposed to light. Addition of the photosensitizer to the already infected cells produced no effect on their intracellular parasites. However, light irradiation lysed these macrophages and also those infected with parasites preincubated with the photosensitizer at a concentration of 5 µg · ml–1 or higher. Photosensitized Leishmania cells are highly susceptible to cytolysis, apparently due to the generation of reactive oxidative species on light illumination, suggestive of inefficiency of their antioxidant mechanisms. Efficient delivery of photosensitizers to intracellular Leishmania is expected to increase their therapeutic potentials against leishmaniasis.

* Corresponding author. Mailing address: Chicago Medical School, 3333 Green Bay Rd., N. Chicago, IL 60064. Phone (847) 578-8837. Fax: (847) 578-3349. E-mail: kwang-poo.chang{at}rosalindfranklin.edu.

Antimicrobial Agents and Chemotherapy, November 2005, p. 4474-4484, Vol. 49, No. 11
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.11.4474-4484.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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