Document Detail


In vitro sensitivity testing of Leishmania clinical field isolates: preconditioning of promastigotes enhances infectivity for macrophage host cells.
MedLine Citation:
PMID:  19752271     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Diagnostic material from patients with leishmaniasis is generally available as promastigotes, and proper testing for susceptibility to first-line drugs by the intracellular amastigote assay is frequently hampered by the poor infectivity of the promastigotes for the macrophage host cell. Several conditions for optimization of the in vitro metacyclogenesis and cell infectivity of Leishmania donovani, L. guyanensis, and L. braziliensis field strains obtained from patients receiving standard antimony medication were investigated. Triggering log-phase promastigotes to become amastigote-like by increasing the temperature or acidifying the culture medium was not successful. Adequate metacyclogenesis and the highest levels of macrophage infection were obtained after 5-day-old late-log-phase promastigote cultures were preconditioned at 25 degrees C to pH 5.4 for 24 h in Schneider's medium prior to infection. The susceptibility assay with primary peritoneal mouse macrophages included pentavalent antimony (Sb(V); sodium stibogluconate), trivalent antimony (Sb(III); potassium antimonyl tartrate), miltefosine, and the experimental drug PX-6518. All strains were sensitive to miltefosine (50% inhibitory concentration [IC(50)] < 10 microM) and PX-6518 (IC(50) < 2 microg/ml) but showed distinct susceptibility to Sb(V) and/or Sb(III), depending on whether they were derived from cured, relapse, or nonresponder patients. Within the available set of Leishmania species and strains, simultaneous Sb(V)-Sb(III) resistance was clearly associated with treatment failure; however, a larger set of isolates is still needed to judge the predictive value of Sb(V)-Sb(III) susceptibility profiling on treatment outcome. In conclusion, the proposed conditioning protocol further contributes toward a more standardized laboratory model for evaluation of the drug sensitivities of field isolates.
Authors:
Raquel Inocêncio da Luz; Marieke Vermeersch; Jean-Claude Dujardin; Paul Cos; Louis Maes
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-09-14
Journal Detail:
Title:  Antimicrobial agents and chemotherapy     Volume:  53     ISSN:  1098-6596     ISO Abbreviation:  Antimicrob. Agents Chemother.     Publication Date:  2009 Dec 
Date Detail:
Created Date:  2009-11-17     Completed Date:  2010-02-01     Revised Date:  2010-09-28    
Medline Journal Info:
Nlm Unique ID:  0315061     Medline TA:  Antimicrob Agents Chemother     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5197-203     Citation Subset:  IM    
Affiliation:
Faculty of Pharmaceutical, Biomedical and Veterinary Sciences, Antwerp University, Antwerp, Belgium.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antimony Sodium Gluconate / pharmacology
Antiprotozoal Agents / pharmacology
Cells, Cultured
Flow Cytometry
Leishmania / drug effects*,  isolation & purification,  pathogenicity*
Leishmaniasis / parasitology*
Macrophages, Peritoneal / parasitology*
Mice
Microscopy
Parasitic Sensitivity Tests
Phosphorylcholine / analogs & derivatives,  therapeutic use
Saponins / pharmacology
Temperature
Triterpenes / pharmacology
Chemical
Reg. No./Substance:
0/Antimony Sodium Gluconate; 0/Antiprotozoal Agents; 0/PX-6518; 0/Saponins; 0/Triterpenes; 107-73-3/Phosphorylcholine; 58066-85-6/miltefosine
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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