Assessment of Leishmania cell lines expressing high levels of beta-galactosidase as alternative tools for the evaluation of anti-leishmanial drug activity.

Autor: da Silva Santos AC; Departamento de Imunologia, Instituto Aggeu Magalhães- FIOCRUZ, Recife, Pernambuco, Brazil. Electronic address: aline.caroline.bm@gmail.com., Moura DMN; Departamento de Imunologia, Instituto Aggeu Magalhães- FIOCRUZ, Recife, Pernambuco, Brazil., Dos Santos TAR; Departamento de Imunologia, Instituto Aggeu Magalhães- FIOCRUZ, Recife, Pernambuco, Brazil., de Melo Neto OP; Departamento de Microbiologia, Instituto Aggeu Magalhães- FIOCRUZ, Recife, Pernambuco, Brazil., Pereira VRA; Departamento de Imunologia, Instituto Aggeu Magalhães- FIOCRUZ, Recife, Pernambuco, Brazil.
Jazyk: angličtina
Zdroj: Journal of microbiological methods [J Microbiol Methods] 2019 Nov; Vol. 166, pp. 105732. Date of Electronic Publication: 2019 Oct 17.
DOI: 10.1016/j.mimet.2019.105732
Abstrakt: Leishmaniasis, caused by protozoa belonging to the genus Leishmania, is an important public health problem found in >90 countries and with still limited options for treatment. Development of new anti-leishmanial drugs is an urgent need and the identification of new active compounds is a limiting factor that can be accelerated through large scale drug screening. This requires multiple steps and can be expensive and time consuming. Here, we propose an alternative approach for the colorimetric assessment of anti-Leishmania drug activity that can be easily scaled up. L. amazonensis and L. infantum cell lines were generated having the β-galactosidase (β-gal) gene integrated into their chromosomal 18S rRNA (ssu) locus. Both cell lines expressed high levels of β-gal and had their growth easily monitored and quantified colorimetrically. These two cell lines were then evaluated as tools to assess drug susceptibility and their use was validated through in vitro assays with Amphotericin B, which is routinely used against leishmaniasis. β-gal expression was also confirmed through flow-cytometry, another method of phenotypic detection. With these recombinant parasites, an alternative in vitro model of drug screening against cutaneous and visceral leishmaniasis is now available.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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