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Clinical isolates of Leishmania, from visceral leishmaniasis (VL) cases in Nepal and from cutaneous leishmaniasis (CL) cases in Peru, were cultured using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to type species and strain. Promastigotes from 38 isolates, within eight passages from isolation, were used to infect mouse peritoneal macrophage cultures in vitro, and the amastigote sensitivity to miltefosine was determined. The concentration required to kill 50% of intracellular amastigotes from Nepalese VL isolates, all typed as Leishmania (L.) donovani (N = 24) from both Sbv responders and nonresponders, ranged from 8.7 to 0.04 µg/mL. In contrast, the concentration required to kill 50% intracellular amastigotes from isolates from Peru, typed as L.(V.) braziliensis (N = 8), was > 30 to 8.4 µg/mL, L.(V.) guyanensis (N = 2) > 30 to 1.9 µg/mL, L.(L.) mexicana (N = 1) > 30 µg/mL, and L. (V.) lainsoni (N = 4) was 3.4 to 1.9 µg/mL. This demonstrates a notable difference in the intrinsic sensitivity of Leishmania species to miltefosine in vitro. If this model can be correlated to therapeutic outcome, it may have implications for the interpretation of clinical trials.
Received June 29, 2004. Accepted for publication February 11, 2005.
Financial support: This study was funded by the European Commission, LEISHNATDRUG-R ICA4-2000-10109.
* Address correspondence to Vanessa Yardley, ITD, LSHTM, Keppel Street, London WC1E 7HT, United Kingdom. E-mail: Vanessa. Yardley{at}lshtm.ac.uk; scroft{at}dndi.org
Authors’ addresses: Vanessa Yardley and Simon Croft, London School of Hygiene & Tropical Medicine (University of London), Department of Infectious and Tropical Diseases, Keppel Street, London WC1E 7HT, UK. Telephone: +44 (0)20 7927-2462, Fax: +44 (0)20 7927-2807, E-mail: vanessa.yardley{at}lshtm.ac.uk, simon.croft{at}lshtm.ac.uk. Simonne de Doncker and Jean-Claude Dujardin, ITMA, Protozoology Unit, Nationalestraat 155, B-2000 Antwerpen, Belgium. Telephone: +32 3 2476358, Fax: +32 3 2476359, E-mail: SDoncker{at}proto.itg.be, jcdujard{at}itg.be. Cesar Miranda and Alejandro Llanos-Cuentas, Universidad Peruana Cayetano Heredia, Insituto de Medicine Tropical "Alexander von Humbolt," P.O. Box 4214, Lima 100, Peru. Telephone: +511 4827739, Fax: +511 3820338, E-mail: cesar.miranda{at}mail.mcgill.ca, allanos{at}upch.edu.pe. Siddhartha Koirala and Suman Rijal, BP Koirala Institute of Health Sciences, Dharan, Nepal. Telephone: +977 25 25555 ext 2052, Fax: +977 25 20251, E-mail: siddharthakoirala2001{at}yahoo.com, sumanrijal2{at}yahoo.com. Francois Chappuis, Travel & Migration Medicine Unit, Geneva University Hospital, 24 Micheli-du-Crest, 1211 Geneva-14, Switzerland. Telephone: +41 22 3729620, Fax: +41 22 3729626, E-mail: francois.chappuis{at}hcuge.ch.
Reprint requests: Vanessa Yardley, ITD, LSHTM, Keppel Street, London WC1E 7HT, UK. Fax: +44 (0)20 7927 2807, E-mail: vanessa.yardley{at}lshtm.ac.uk.
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