Parasite burden in Leishmania (Leishmania) amazonensis-infected mice: Validation of luciferase as a quantitative tool

Autor(es)

Reimao, Juliana Q. ^[1] ; Trinconi, Cristiana T. ^[1] ; Yokoyama-Yasunaka, Jenicer K. ^[1] ; Miguel, Danilo C. ^[1] ; Kalil, Sandra P. ^[1] ; Uliana, Silvia R. B. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Ciencias Biomed, Dept Parasitol, BR-05508000 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Journal of Microbiological Methods; v. 93, n. 2, p. 95-101, MAY 2013.
• Citações Web of Science: 23

Resumo

Given the lack of effective and safe alternatives to the drugs already in use, considerable efforts are being applied to the search of new therapeutic options to treat leishmaniasis. A necessary step in the discovery of antileishmanial drugs is the validation of drug candidates in mouse models. The standard methods to quantify the parasite burden in animal models, mainly culture-based, are time consuming and expensive. In recent years, in vivo imaging systems have been proposed as a tool to overcome these problems, allowing paraSite detection in living organisms. Here we compared different treatment efficacy evaluation approaches. Recombinant Leishmania (L) amazonensis lines expressing the luciferase gene (La-LUC) were obtained and characterized for biological properties as compared with the wild type (WT) parental line. Bioluminescence generated by La-LUC was shown to correlate with the number of promastigotes in vitro. La-LUC promastigotes and intracellular amastigotes were equally sensitive to amphotericin B (AmB) as the WT parasites. The clinical pattern of lesion development upon infection with the transgenic lines was similar to lesions observed after infection with the WT strain. The half maximal effective dose (ED50) of AmB was determined in La-LUC infected mice through quantification of bioluminescence in vivo and ex vivo, by limiting dilution and using clinical parameters. There was agreement in the ED50 determined by all methods. Quantification of bioluminescence in vivo and/or ex vivo was elected as the best tool for determining parasite burden to assess drug efficacy in infected mice. Furthermore, the detailed analysis of AmB effectiveness in this model generated useful data to be used in drug combination experiments. (C) 2013 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 11/20484-7 - Tamoxifeno no tratamento de leishmaniose: avaliação de eficácia em esquemas de combinação de drogas e estudo do mecanismo de ação
• Beneficiário: Silvia Reni Bortolin Uliana
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 11/18858-6 - Tamoxifeno como droga anti-leishmania: atividade em esquemas terapêuticos combinados e estudo do mecanismo de ação
• Beneficiário: Cristiana de Melo Trinconi Tronco
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 11/21970-2 - Moduladores seletivos de receptores de estrógeno como candidatos a fármacos para leishmaniose visceral: avaliação de associações de fármacos e investigação de mecanismos de ação leishmanicida
• Beneficiário: Juliana Quero Reimão Dalla Zanna
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado