A new fluorometric method for anti-Leishmania drug screening on axenic amastigotes

• Autorzy: Le Pape P , Pagniez F , Abdala-Valencia H
• Języki publikacji: EN

Abstrakty

EN

Primary screening of possible antileishmanial compounds is currently based either on the use of promastigotes, which are not the clinically relevant stage, or on the time-consuming intracellular amastigote test. With the aim of discovering new antileishmanial derivatives, a semi-automatized method using Leishmania axenic amastigotes and a fluorometric growth indicator (Alamar Blue®) was developed. This assay was evaluated in a comparative study of the susceptibility of promastigotes, axenic amastigotes and intracellular amastigotes of Leishmania mexicana with standard drugs used in antileishmanial therapy (meglumine antimoniate, amphotericin B, ketoconazole and allopurinol) and recently developed ether lipid compounds (edelfosine and miltefosine). The results showed that while inhibitory concentration 50s (IC₅₀s) of axenic and intracellular amastigotes were similar for all of the drugs tested, promastigotes and intracellular amastigotes IC₅₀s differed for three of the six compounds. In conclusion, this axenic amastigote model allowed us to combine the ease of in vitro drug screening on promastigote with the reliability of the IC₅₀ determination on intracellular amastigotes.

Słowa kluczowe

EN

axenic amastigote; Leishmania mexicana; antileishmanial therapy; drug; fluorometric method; promastigote; screening test

• Wydawca: -
• Czasopismo: Acta Parasitologica
• Rocznik: 2003
• Tom: 48
• Numer: 4
• Opis fizyczny: p.301-305,fig.,ref.

Twórcy

autor

Le Pape P

• Laboratoire de Parasitologie et de Mycologie Medicale, UPRES EA 1155, Faculte de Pharmacie, Universite de Nantes, 1 rue Gaston Veil, 44035 Nantes cedex, France

autor

Pagniez F

• Laboratoire de Parasitologie et de Mycologie Medicale, UPRES EA 1155, Faculte de Pharmacie, Universite de Nantes, 1 rue Gaston Veil, 44035 Nantes cedex, France

autor

Abdala-Valencia H

• Laboratoire de Parasitologie et de Mycologie Medicale, UPRES EA 1155, Faculte de Pharmacie, Universite de Nantes, 1 rue Gaston Veil, 44035 Nantes cedex, France

Bibliografia

• Abdala H., Robert J.M., Le Pape P., Wielgosz G., Robert-Piessard S., Le Baut G. 2000. Synthesis and antileishmanial activity of new l-(pyridin-2-yl)imidazolidin-2-one derived from 2-amino-4,6-dimethylpyridine. Arzneimittel-Forschung, 50, 479-484.
• Ahmed S.A., Gogal Jr. R.M., Walsh J.E. 1994. A new rapid and simple non-radioactive assay to monitor and determine the proliferation of lymphocytes: an alternative to H3-thymidine incorporation assay. Journal of Immunological Methods, 170, 211-224.
• Baker C.N., Tenover F.C. 1996. Evaluation of Alamar colorimetric broth microdilution susceptibility testing method for staphylococci and enterococci. Journal of Clinical Microbiology, 34, 2654-2659.
• Bates P.A. 1994. Complete developmental cycle of Leishmania mexicana in axenic culture. Parasitology, 108,1-9.
• Bates P.A., Robertson C.D., Tetley L., Coombs G.H. 1992. Axenic cultivation and characterization of Leishmania mexicana amastigote-like forms. Parasitology, 105, 193-202.
• Berg K., Zhai L., Chen M., Kharmzmi A., Owen T. 1994. The use of a water-soluble formazan complex to quantitate the cell number and mitochondrial function of Leishmania major promastigotes. Parasitology Research, 80, 235-239.
• Berman J.D. 1997. Human leishmaniasis: clinical, diagnostic and chemotherapeutic developments in the last 10 years. Clinical Infectious Diseases, 24, 684-703.
• Bodley A.L., McGarry M.W., Shapiro T.A. 1995. Drug cytotoxicity for African trypanosomes and Leishmania species. Journal of Infectious Diseases, 172, 1157-1159.
• Callahan H.L., Portal A.C., Devereaux R., Grogl M. 1997. An axenic amastigote system for drug screening. Antimicrobial Agents and Chemotherapy, 41, 818-822.
• Croft S.L., Snowdon D., Yardley V. 1996. The activity of four anticancer alkylphospholipid against L. donovani, T. cruzi and 71 brucei. Journal of Antimicrobial Chemotherapy, 38, 1041-1047.
• Jahn B., Stuben A., Bhakdi S. 1996. Colorimetric susceptibility testing for Aspergillus fumigatus: comparison of menadione-augmented 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide and Alamar blue tests. Journal of Clinical Microbiology, 34, 2039-2041.
• Kaminsky R., Bran R. 1993. In vitro assays to determine drug sensitivities of African trypanosomiasis: a review. Acta Tropica, 54, 279-289.
• Le Pape P, Pagniez F., Abdala H. 2002. A new automatized fluorometric assay for anti-Leishmania drug screening on promastigotes. Acta Parasitologica, 47, 79-81.
• Mikus J., Steverding D. 2000. A simple colorimetric method to screen drug cytotoxicity against Leishmania using the dye Alamar Blue®. Parasitology International, 48, 265-269.
• Mosmann T. 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 6, 55-63.
• Pagniez F, Le Pape P. 2001. New fluorometric screening test for possible antifungal drugs. Journal de Mycologie Médicale, 11, 73-78.
• Sereno D., Cavaleyro M., Zemzoumi K., Maquaire S., Ouaissi A., Lemesre J.L. 1998. Axenically grown amastigotes of Leishmania infantum used as an in vitro model to investigate the pentavalent antimony mode of action. Antimicrobial Agents and Chemotherapy, 42, 3097-3102.
• Sereno D., Lemesre J.L. 1997. Use of an enzymatic micromethod to quantify amastigote stage of Leishmania amazonensis in vitro. Parasitology Research, 83, 401-403.
• Tiballi R.N., He X., Zarins L.T., Revankar S.G., Kauffman C.A. 1995. Use of a colorimetric system for yeast susceptibility testing. Journal of Clinical Microbiology, 33, 915-917.
• Yajko D.M., Madej J.J., Lancaster M.V., Sanders C.A., Cawthon V.L., Gee B., Babst A., Hadley W.K. 1995. Colorimetric method for determining MICs of antimicrobial agents for Mycobacterium tuberculosis. Journal of Clinical Microbiology, 33,2324-2327.