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Studies in vitro on infectivity and sensitivity to antileishmanial drugs in New World Leishmania species transfected with the green fluorescent protein [pIR3(-)-eGFP]

Published online by Cambridge University Press:  23 August 2017

GENESIS PALACIOS ,
ADRIANA PARODI ,
YULIETH A. UPEGUI ,
ANDRES MONTOYA ,
SERGIO PULIDO ,
IVÁN D. VÉLEZ  and
SARA M. ROBLEDO Open the ORCID record for SARA M. ROBLEDO [Opens in a new window]
GENESIS PALACIOS
Affiliation:
PECET-Facultad de Medicina, Universidad de Antioquia, Medellin, Colombia
ADRIANA PARODI
Affiliation:
PECET-Facultad de Medicina, Universidad de Antioquia, Medellin, Colombia
YULIETH A. UPEGUI
Affiliation:
PECET-Facultad de Medicina, Universidad de Antioquia, Medellin, Colombia
ANDRES MONTOYA
Affiliation:
PECET-Facultad de Medicina, Universidad de Antioquia, Medellin, Colombia
SERGIO PULIDO
Affiliation:
PECET-Facultad de Medicina, Universidad de Antioquia, Medellin, Colombia
IVÁN D. VÉLEZ
Affiliation:
PECET-Facultad de Medicina, Universidad de Antioquia, Medellin, Colombia
SARA M. ROBLEDO*
Affiliation:
PECET-Facultad de Medicina, Universidad de Antioquia, Medellin, Colombia
*
*Corresponding author. PECET-Facultad de Medicina, Universidad de Antioquia, Calle 70 # 52-21, Medellin, Colombia. E-mail: [email protected]
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Summary

Current chemotherapeutic agents for leishmaniasis have several disadvantages interfering with the effective treatment and therefore more and better antileishmanial drugs are needed. Discovery of candidates for leishmaniasis treatment requires not only accurate and precise methodologies but also well-known biological system to measure infectivity of parasites and antileishmanial activity of the new compounds. Significant variation in the in vitro and in vivo infectivity and sensitivity to established and experimental drugs in Leishmania strains are reported. This work reports the in vitro biological behavior and antileishmanial drugs sensitivity of different green fluorescent protein transfectant Leishmanias strains. The in vitro growth kinetic and infectivity to U937 cells vary slightly in the Leishmania transfectant strains in comparison with their correspondant wild-type. However, the insertion of the pIR3(-)-eGFP may affect the sensitivity of the parasites to meglumine antimoniate (MA) and miltefosine but not to amphotericin B (AMB) and pentamidine isethionate. In consequence, AMB or pentamidine isethionate but not MA or miltefosine should be used as antileishmanial control drugs during in vitro assays of antileishmanial activity. Furthermore, is recommended to test compounds against more than one Leishmania strain in order to verify that the antileihmanial activity of these compound is similar among species.

Keywords

LeishmanialeishmaniasisL. panamensisL. guyanensisL.braziliensisL.mexicanaL.amazonensisinfectivitydrug sensitivity
Type
Research Article
Information
Parasitology , Volume 144 , Issue 13 , November 2017 , pp. 1718 - 1725
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Copyright
Copyright © Cambridge University Press 2017 

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Footnotes

†

These authors contributed equally to this paper.

References

REFERENCES

Allison, D. G. and Sattenstall, M. A. (2007). The influence of green fluorescent protein incorporation on bacterial physiology: a note of caution. Journal of Applied Microbiology 103, 318–324.CrossRefGoogle ScholarPubMed
Bertholet, S. and Mauël, J. (2000). Human monocytic U937 cells transfected with human hepatic inducible nitric oxide synthase exhibit leishmanicidal activity. Journal of Leukocyte Biology 67, 34–39.CrossRefGoogle ScholarPubMed
Bolhassani, A., Taheri, T., Taslimi, Y., Zamanilui, S., Zahedifard, F., Seyed, N., Torkashvand, F., Vaziri, B. and Rafati, S. (2011). Fluorescent Leishmania species: development of stable GFP expression and its application for in vitro and in vivo studies. Experimental Parasitology 127, 637–645. doi: 10.1016/j.exppara.2010.12.006.CrossRefGoogle ScholarPubMed
Buckner, F. S. and Wilson, A. J. (2005). Colorimetric assay for screening compounds against Leishmania amastigotes grown in macrophages. American Journal of Tropical Medicine and Hygiene 72, 600–605.CrossRefGoogle ScholarPubMed
Buckner, F. S., Verlinde, C. L., La Flamme, A. C. and Van Voorhis, W. C. (1996). Efficient technique forscreening drugs for activity against Trypanosoma cruzi using parasites expressingbeta-galactosidase. Antimicrobial Agents and Chemotherapy 40, 2592–2597.CrossRefGoogle Scholar
Da Silva, I. A. Jr, Morato, C. I., Quixabeira, V. B., Pereira, L. I., Dorta, M. L., de Oliveira, M. A., Horta, M. F. and Ribeiro-Dias, F. (2015). In vitro metacyclogenesis of Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis clinical field isolates, as evaluated by morphology, complement resistance, and infectivity to humanmacrophages. BioMed Research International 2015, 393049. doi: 10.1155/2015/393049.CrossRefGoogle Scholar
Finney, D. J. (1978) Statistical Method in Biological Assay, p. 508, 3rd edn. Charles Griffin & Co., London and High Wycombe.Google Scholar
Lang, T., Goyard, S., Lebastard, M. and Milon, G. (2005). Bioluminescent Leishmania expressing luciferase for rapid and high throughput screening of drugs acting on amastigote-harbouring macrophages and for quantitative real-time monitoring of parasitism features in living mice. Cell Microbiology 7, 383–392.CrossRefGoogle ScholarPubMed
Liu, H. S., Jan, M. S., Chou, C. K., Chen, P. H. and Ke, N. J. (1999). Is green fluorescent protein toxic to the living cells? Biochemical and Biophysical Research Communications 260, 712–717.CrossRefGoogle Scholar
Misra, S., Tripathi, M. K. and Chaudhuri, G. (2005). Down-regulation of 7SL RNA expression andimpairment of vesicular protein transport pathways by Leishmania infection ofmacrophages. Journal of Biological Chemistry 280, 29364–29373.CrossRefGoogle Scholar
Ovalle-Bracho, C., Franco-Muñoz, C., Londoño-Barbosa, D., Restrepo-Montoya, D. and Clavijo-Ramírez, C. (2015). Changes in Macrophage Gene Expression Associated with Leishmania (Viannia) braziliensis Infection. PLoS ONE 10, e0128934. doi: 10.1371/journal.pone.0128934.CrossRefGoogle ScholarPubMed
Puentes, F., Diaz, D., Hoya, R. D., Gutíerrez, J. A., Lozano, J. M., Patarroyo, M. E. and Moreno, A. (2000). Cultivation and characterization of stable Leishmania guyanensis complex axenic amastigotes derived from infected U937 cells. American Journal of Tropical Medicine and Hygiene 63, 102–110.CrossRefGoogle ScholarPubMed
Pulido, S. A., Munoz, D. L., Restrepo, A. M., Mesa, C. V., Alzate, J. F., Vélez, I. D. and Robledo, S. M. (2012). Improvement of the green fluorescent protein reporter system in Leishmania spp. for the in vitro and in vivo screening of antileishmanial drugs. Acta Tropica 122, 36–45. doi: 10.1016/j.actatropica.2011.11.015.CrossRefGoogle ScholarPubMed
Robledo, S. M., Valencia, A. Z. and Saravia, N. G. (1999). Sensitivity to Glucantime of Leishmania Viannia isolated from patients prior to treatment. Journal of Parasitology 85, 360–366.CrossRefGoogle ScholarPubMed
Roy, G., Dumasm, C., Serenom, D., Wum, Y., Singhm, A. K., Tremblay, M. J., Ouellette, M., Olivier, M. and Papadopoulou, B. (2000). Episomal and stable expression of the luciferase reporter gene for quantifying Leishmania spp. infections in macrophages and in animal models. Molecular and Biochemical Parasitology 110, 195–206.CrossRefGoogle ScholarPubMed
Tao, W., Evans, B. G., Yao, J., Cooper, S., Cornetta, K., Ballas, C. B., Hangoc, G. and Broxmeyer, H. E. (2007). Enhanced green fluorescent protein is a nearly ideal long-term expression tracer for hematopoietic stem cells, whereas DsRed-express fluorescent protein is not. Stem Cells 25, 670–678.CrossRefGoogle ScholarPubMed
Taylor, V. M., Muñoz, D. L., Cedeño, D. L., Vélez, I. D., Jones, M. A. and Robledo, S. M. (2010). Leishmania tarentolae: utility as an in vitro model for screening of antileishmanial agents. Experimental Parasitology 126, 471–475. doi: 10.1016/j.exppara.2010.05.016.CrossRefGoogle Scholar
World Health Organization. (2010). Control of the leishmaniasis: report of a meeting of the WHO Expert Committee on the Control of Leishmaniases. [Online] Available from: http://apps.who.int/iris/bitstream/10665/44412/1/WHO_TRS_949_eng.pdf [Accessed on 10th Sep, 2016].Google Scholar