In Vivo Bioluminescent Monitoring of Parasites in BALB/c Mouse Models of Cutaneous Leishmaniasis Drug Discovery

  • Diana CaridhaEmail author
  • Susan Leed
  • Alicia Cawlfield
Part of the Methods in Molecular Biology book series (MIMB, volume 2081)


Confirming the in vivo efficacy of potential antileishmanial compounds that display in vitro potency and good chemical characteristics is one of the most important steps in preclinical research drug discovery before human clinical trials begin. Here we describe the use of the in vivo bioluminescent monitoring of high and low inocula of luciferase-expressing Leishmania major (L. major) parasites in traditional and more innovative rodent models of in vivo cutaneous leishmaniasis (CL) drug discovery.

Key words

Cutaneous leishmaniasis (CL) Rodent models In vivo imaging IVIS Bioluminescence signal Luciferase-expressing Leishmania major Drug efficacy Base of the tail Footpad Ear infections 



Disclaimer: Material has been reviewed by the Walter Reed Army Institute of Research. There is no objection to its presentation and/or publication. The opinions or assertions contained herein are the private views of the author, and are not to be construed as official, or as reflecting true views of the Department of the Army or the Department of Defense.

Research was conducted under an approved animal use protocol in an AAALACi accredited facility in compliance with the Animal Welfare Act and other federal statutes and regulations relating to animals and experiments involving animals and adheres to principles stated in the Guide for the Care and Use of Laboratory Animals, NRC Publication, 2011 edition.


  1. 1.
    World Health Organization (2010) Control of the leishmaniases. World Health Organ Tech Rep Ser 949:xii–xiii 1-186Google Scholar
  2. 2.
    Croft SL, Seifert K, Yardley V (2006) Current scenario of drug development for leishmaniasis. Indian J Med Res 123(3):399–410PubMedGoogle Scholar
  3. 3.
    Pigott DM, Bhatt S, Golding N et al (2014) Global distribution maps of the leishmaniases. Elife 3.
  4. 4.
    Mears ER, Modabber F, Don R, Johnson GE (2015) A review: the current in vivo models for the discovery and utility of new anti-leishmanial drugs targeting cutaneous leishmaniasis. PLoS Negl Trop Dis 9(9):e0003889. Scholar
  5. 5.
    Robledo SM, Carrillo LM, Daza A et al (2012) Cutaneous leishmaniasis in the dorsal skin of hamsters: a useful model for the screening of antileishmanial drugs. J Vis Exp 62:3533. Scholar
  6. 6.
    Milon G, Del Giudice G, Louis JA (1995) Immunobiology of experimental cutaneous leishmaniasis. Parasitol Today 11(7):244–247CrossRefGoogle Scholar
  7. 7.
    Gomes-Silva A, Valverde JG, Ribeiro-Romao RP et al (2013) Golden hamster (Mesocricetus auratus) as an experimental model for Leishmania (Viannia) braziliensis infection. Parasitology 140(6):771–779. Scholar
  8. 8.
    Ribeiro-Romao RP, Moreira OC, Osorio EY et al (2014) Comparative evaluation of lesion development, tissue damage, and cytokine expression in golden hamsters (Mesocricetus auratus) infected by inocula with different Leishmania (Viannia) braziliensis concentrations. Infect Immun 82(12):5203–5213. Scholar
  9. 9.
    Kimblin N, Peters N, Debrabant A et al (2008) Quantification of the infectious dose of Leishmania major transmitted to the skin by single sand flies. Proc Natl Acad Sci U S A 105(29):10125–10130. Scholar
  10. 10.
    Abdeladhim M, Kamhawi S, Valenzuela JG (2014) What’s behind a sand fly bite? The profound effect of sand fly saliva on host hemostasis, inflammation and immunity. Infect Genet Evol 28:691–703. Scholar
  11. 11.
    Belkaid Y, Kamhawi S, Modi G et al (1998) Development of a natural model of cutaneous leishmaniasis: powerful effects of vector saliva and saliva preexposure on the long-term outcome of Leishmania major infection in the mouse ear dermis. J Exp Med 188(10):1941–1953CrossRefGoogle Scholar
  12. 12.
    Cawlfield A, Vesely B, Ngundam F et al (2018) Use of in vivo imaging system technology in Leishmania major BALB/c mouse ear infection studies. J Med Entomol 55(2):429–435. Scholar
  13. 13.
    Greer LF 3rd, Szalay AA (2002) Imaging of light emission from the expression of luciferases in living cells and organisms: a review. Luminescence 17(1):43–74. Scholar
  14. 14.
    Lang T, Goyard S, Lebastard M, 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 Microbiol 7(3):383–392. Scholar
  15. 15.
    Roy G, Dumas C, Sereno D et al (2000) Episomal and stable expression of the luciferase reporter gene for quantifying Leishmania spp. infections in macrophages and in animal models. Mol Biochem Parasitol 110(2):195–206CrossRefGoogle Scholar
  16. 16.
    Thalhofer CJ, Graff JW, Love-Homan L et al (2010) In vivo imaging of transgenic Leishmania parasites in a live host. J Vis Exp 41:1980. Scholar
  17. 17.
    Caridha D, Parriot S, Hudson TH et al (2017) Use of optical imaging technology in the validation of a new, rapid, cost-effective drug screen as part of a tiered in vivo screening paradigm for development of drugs to treat cutaneous leishmaniasis. Antimicrob Agents Chemother 61(4):e02048-16. Scholar
  18. 18.
    Schuster S, Hartley MA, Tacchini-Cottier F, Ronet C (2014) A scoring method to standardize lesion monitoring following intra-dermal infection of Leishmania parasites in the murine ear. Front Cell Infect Microbiol 4:67. Scholar
  19. 19.
    Lecoeur H, Buffet PA, Milon G, Lang T (2010) Early curative applications of the aminoglycoside WR279396 on an experimental Leishmania major-loaded cutaneous site do not impair the acquisition of immunity. Antimicrob Agents Chemother 54(3):984–990. Scholar
  20. 20.
    Lecoeur H, Buffet P, Morizot G et al (2007) Optimization of topical therapy for Leishmania major localized cutaneous leishmaniasis using a reliable C57BL/6 model. PLoS Negl Trop Dis 1(2):e34. Scholar
  21. 21.
    Sacks DL, Melby PC (2001) Animal models for the analysis of immune response to leishmaniasis. Curr Protoc Immunol . Chapter 19: Unit 19.2.
  22. 22.
    Sacks DL, Melby PC (2015) Animal models for the analysis of immune response to leishmaniasis. Curr Protoc Immunol 108(19):2): 1–2):24. Scholar
  23. 23.
    Fischer MR, Dominik J, Kautz-Neu K et al (2013) Animal model for cutaneous leishmaniasis. In: Has C, Sitaru C (eds) Molecular dermatology. Methods in molecular biology (methods and protocols), vol 961. Humana Press, Totowa, NJ. Scholar
  24. 24.
    Lawyer P, Killick-Kendrick M, Rowland T et al (2017) Laboratory colonization and mass rearing of phlebotomine sand flies (Diptera, Psychodidae). Parasite 24:42. Scholar
  25. 25.
    Baldwin TM, Elso C, Curtis J et al (2003) The site of Leishmania major infection determines disease severity and immune responses. Infect Immun 71(12):6830–6834. Scholar
  26. 26.
    Spath GF, Beverley SM (2001) A lipophosphoglycan-independent method for isolation of infective Leishmania metacyclic promastigotes by density gradient centrifugation. Exp Parasitol 99(2):97–103CrossRefGoogle Scholar
  27. 27.
    Turner PV, Brabb T, Pekow C, Vasbinder MA (2011) Administration of substance to laboratory animals: routes of administration and factors to consider. J Am Assoc Lab Anim Sci 50(5):600–613PubMedPubMedCentralGoogle Scholar
  28. 28.
    Nasseri M, Modabber FZ (1979) Generalized infection and lack of delayed hypersensitivity in BALB/c mice infected with Leishmania tropica major. Infect Immun 26(2):611–614PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Walter Reed Army Institute of ResearchSilver SpringUSA
  2. 2.Dwight D. Eisenhower Army Medical CenterFort GordonUSA

Personalised recommendations