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In Vitro Infections of Macrophage-Like Cell Lines with Leishmania infantum for Drug Screening

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Leishmania

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1971))

Abstract

The study of in vitro infections is essential to evaluate distinct aspects of Leishmania biology and also invaluable for more meaningful in vitro screening of promising chemical entities. Macrophage-like cells lines from different origins are amenable to Leishmania infection. Cell lines due to their stability and standardization potential are highly valued for their capacity to support reproducible infections and consistent data. In fact, these cells have been a mainstay of leishmaniasis research for more than 40 years. In this context, the human monocytic THP-1 cell line is commonly used as it can be differentiated with phorbol-12myristate-13-acetate (PMA) into macrophages that are susceptible to Leishmania infection. In this section, we will describe generalities concerning the use of cell lines for in vitro Leishmania infection using THP-1 derived macrophages and Leishmania infantum axenic amastigotes expressing luciferase associated to preclinical drug screening as example.

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References

  1. WHO (2017) Leishmaniasis. http://www.who.int/leishmaniasis/en/. Accessed 30 Nov 2017.

  2. Edmondson R, Broglie JJ, Adcock AF, Yang L (2014) Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors. Assay Drug Dev Technol 12(4):207–218. https://doi.org/10.1089/adt.2014.573

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Zulfiqar B, Shelper TB, Avery VM (2017) Leishmaniasis drug discovery: recent progress and challenges in assay development. Drug Discov Today 22(10):1516–1531. https://doi.org/10.1016/j.drudis.2017.06.004

    Article  CAS  PubMed  Google Scholar 

  4. De Muylder G, Ang KK, Chen S, Arkin MR, Engel JC, McKerrow JH (2011) A screen against Leishmania intracellular amastigotes: comparison to a promastigote screen and identification of a host cell-specific hit. PLoS Negl Trop Dis 5(7):e1253. https://doi.org/10.1371/journal.pntd.0001253

    Article  PubMed  PubMed Central  Google Scholar 

  5. Nuhs A, De Rycker M, Manthri S, Comer E, Scherer CA, Schreiber SL, Ioset JR, Gray DW (2015) Development and validation of a novel Leishmania donovani screening cascade for high-throughput screening using a novel axenic assay with high predictivity of leishmanicidal intracellular activity. PLoS Negl Trop Dis 9(9):e0004094. https://doi.org/10.1371/journal.pntd.0004094

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Shafi S, Afrin F, Islamuddin M, Chouhan G, Ali I, Naaz F, Sharma K, Zaman MS (2016) Beta-nitrostyrenes as potential anti-leishmanial agents. Front Microbiol 7:1379. https://doi.org/10.3389/fmicb.2016.01379

    Article  PubMed  PubMed Central  Google Scholar 

  7. Santarem N, Cunha J, Silvestre R, Silva C, Moreira D, Ouellette M, Cordeiro-da-Silva A (2014) The impact of distinct culture media in Leishmania infantum biology and infectivity. Parasitology 141(2):192–205. https://doi.org/10.1017/S0031182013001388

    Article  PubMed  Google Scholar 

  8. Maia C, Rolao N, Nunes M, Goncalves L, Campino L (2007) Infectivity of five different types of macrophages by Leishmania infantum. Acta Trop 103(2):150–155. https://doi.org/10.1016/j.actatropica.2007.06.001

    Article  CAS  PubMed  Google Scholar 

  9. Tsuchiya S, Yamabe M, Yamaguchi Y, Kobayashi Y, Konno T, Tada K (1980) Establishment and characterization of a human acute monocytic leukemia cell line (THP-1). Int J Cancer 26(2):171–176

    Article  CAS  PubMed  Google Scholar 

  10. Ogunkolade BW, Colomb-Valet I, Monjour L, Rhodes-Feuillette A, Abita JP, Frommel D (1990) Interactions between the human monocytic leukaemia THP-1 cell line and Old and New World species of Leishmania. Acta Trop 47(3):171–176

    Article  CAS  PubMed  Google Scholar 

  11. Sundstrom C, Nilsson K (1976) Establishment and characterization of a human histiocytic lymphoma cell line (U-937). Int J Cancer 17(5):565–577

    Article  CAS  PubMed  Google Scholar 

  12. Gupta S, Yardley V, Vishwakarma P, Shivahare R, Sharma B, Launay D, Martin D, Puri SK (2015) Nitroimidazo-oxazole compound DNDI-VL-2098: an orally effective preclinical drug candidate for the treatment of visceral leishmaniasis. J Antimicrob Chemother 70(2):518–527. https://doi.org/10.1093/jac/dku422

    Article  CAS  PubMed  Google Scholar 

  13. Ralph P, Moore MA, Nilsson K (1976) Lysozyme synthesis by established human and murine histiocytic lymphoma cell lines. J Exp Med 143(6):1528–1533

    Article  CAS  PubMed  Google Scholar 

  14. Pratt DM, David JR (1982) Monoclonal antibodies recognizing determinants specific for the promastigote state of Leishmania mexicana. Mol Biochem Parasitol 6(5):317–327

    Article  CAS  PubMed  Google Scholar 

  15. Kolodziej H, Kiderlen AF (2005) Antileishmanial activity and immune modulatory effects of tannins and related compounds on Leishmania parasitised RAW 264.7 cells. Phytochemistry 66(17):2056–2071. https://doi.org/10.1016/j.phytochem.2005.01.011

    Article  CAS  PubMed  Google Scholar 

  16. Raschke WC, Baird S, Ralph P, Nakoinz I (1978) Functional macrophage cell lines transformed by Abelson leukemia virus. Cell 15(1):261–267

    Article  CAS  PubMed  Google Scholar 

  17. Nandan D, Lo R, Reiner NE (1999) Activation of phosphotyrosine phosphatase activity attenuates mitogen-activated protein kinase signaling and inhibits c-FOS and nitric oxide synthase expression in macrophages infected with Leishmania donovani. Infect Immun 67(8):4055–4063

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Wellman ML, Krakowka S, Jacobs RM, Kociba GJ (1988) A macrophage-monocyte cell line from a dog with malignant histiocytosis. In Vitro Cell Dev Biol 24(3):223–229

    Article  CAS  PubMed  Google Scholar 

  19. Looker DL, Martinez S, Horton JM, Marr JJ (1986) Growth of Leishmania donovani amastigotes in the continuous human macrophage cell line U937: studies of drug efficacy and metabolism. J Infect Dis 154(2):323–327

    Article  CAS  PubMed  Google Scholar 

  20. Lund ME, To J, O'Brien BA, Donnelly S (2016) The choice of phorbol 12-myristate 13-acetate differentiation protocol influences the response of THP-1 macrophages to a pro-inflammatory stimulus. J Immunol Methods 430:64–70. https://doi.org/10.1016/j.jim.2016.01.012

    Article  CAS  PubMed  Google Scholar 

  21. Borsari C, Santarem N, Torrado J, Olias AI, Corral MJ, Baptista C, Gul S, Wolf M, Kuzikov M, Ellinger B, Witt G, Gribbon P, Reinshagen J, Linciano P, Tait A, Costantino L, Freitas-Junior LH, Moraes CB, Bruno Dos Santos P, Alcantara LM, Franco CH, Bertolacini CD, Fontana V, Tejera Nevado P, Clos J, Alunda JM, Cordeiro-da-Silva A, Ferrari S, Costi MP (2017) Methoxylated 2′-hydroxychalcones as antiparasitic hit compounds. Eur J Med Chem 126:1129–1135. https://doi.org/10.1016/j.ejmech.2016.12.017

    Article  CAS  PubMed  Google Scholar 

  22. ATCC THP1(ATCC TIB-202). https://www.lgcstandards-atcc.org/products/all/TIB-202.aspx. Accessed 4 Jan 2018.

  23. Seifert K, Escobar P, Croft SL (2010) In vitro activity of anti-leishmanial drugs against Leishmania donovani is host cell dependent. J Antimicrob Chemother 65(3):508–511. https://doi.org/10.1093/jac/dkp500

    Article  CAS  PubMed  Google Scholar 

  24. Sereno D, Lemesre JL (1997) Axenically cultured amastigote forms as an in vitro model for investigation of antileishmanial agents. Antimicrob Agents Chemother 41(5):972–976

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Aldo PB, Craveiro V, Guller S, Mor G (2013) Effect of culture conditions on the phenotype of THP-1 monocyte cell line. Am J Reprod Immunol 70(1):80–86. https://doi.org/10.1111/aji.12129

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Wenger SL, Senft JR, Sargent LM, Bamezai R, Bairwa N, Grant SG (2004) Comparison of established cell lines at different passages by karyotype and comparative genomic hybridization. Biosci Rep 24(6):631–639. https://doi.org/10.1007/s10540-005-2797-5

    Article  CAS  PubMed  Google Scholar 

  27. Briske-Anderson MJ, Finley JW, Newman SM (1997) The influence of culture time and passage number on the morphological and physiological development of Caco-2 cells. Proc Soc Exp Biol Med 214(3):248–257

    Article  CAS  PubMed  Google Scholar 

  28. Moreira D, Santarem N, Loureiro I, Tavares J, Silva AM, Amorim AM, Ouaissi A, Cordeiro-da-Silva A, Silvestre R (2012) Impact of continuous axenic cultivation in Leishmania infantum virulence. PLoS Negl Trop Dis 6(1):e1469. https://doi.org/10.1371/journal.pntd.0001469

    Article  PubMed  PubMed Central  Google Scholar 

  29. Ali KS, Rees RC, Terrell-Nield C, Ali SA (2013) Virulence loss and amastigote transformation failure determine host cell responses to Leishmania mexicana. Parasite Immunol 35(12):441–456. https://doi.org/10.1111/pim.12056

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by funds from the Fundação para a Ciência e Tecnologia (FCT)/Ministério da Educação e Ciência (MEC) cofunded by the European Regional Development Fund (FEDER) under the Partnership agreement PT2020, through the Research Unit No.4293. This work also received funds from project POCI-01-0145-FEDER-031013 financed by Portugal 2020, under the Programa Operacional Competitividade e Internacionalização (COMPETE 2020) and Norte-01-0145-FEDER-000012—Structured program on bioengineered therapies for infectious diseases and tissue regeneration, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the FEDER. J.T. is an Investigator FCT funded by National funds through FCT and cofunded through European Social Fund within the Human Potential Operating Programme.

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Correspondence to Anabela Cordeiro-da-Silva .

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Santarém, N., Tavares, J., Cordeiro-da-Silva, A. (2019). In Vitro Infections of Macrophage-Like Cell Lines with Leishmania infantum for Drug Screening. In: Clos, J. (eds) Leishmania. Methods in Molecular Biology, vol 1971. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9210-2_14

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  • DOI: https://doi.org/10.1007/978-1-4939-9210-2_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9209-6

  • Online ISBN: 978-1-4939-9210-2

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