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Leishmania pp 289-301 | Cite as

Quantification of Leishmania Parasites in Murine Models of Visceral Infection

  • Joana Tavares
  • Nuno Santarém
  • Anabela Cordeiro-da-SilvaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1971)

Abstract

Visceral leishmaniasis (VL) is mainly caused by Leishmania donovani (India and East Africa), and Leishmania infantum (Mediterranean Basin and South America) infections. Although murine models of visceral infection lack the clinicopathological aspects of VL in humans, they have been proven useful at advancing our knowledge in the Leishmania field. Indeed, these models have been used not only to better understand the pathophysiology of the infection but also in drug and vaccine development. This chapter focuses on the protocols used to experimentally infect mice and to quantify parasite burdens in mice infected with L. infantum using limiting dilution methodology of target organs and whole-mouse in vivo imaging.

Key words

Leishmania infantum Visceral infection Limiting dilution Whole-mouse in vivo imaging 

Notes

Acknowledgments

We apologize to many researchers in this field whose work we have not been able to cite directly owing to space limitation. 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 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 and the project POCI-01-0145-FEDER-031013 financed by Portugal 2020, under the Programa Operacional Competitividade e Internacionalização (COMPETE 2020). J.T. is an Investigator FCT funded by National funds through FCT and cofunded through European Social Fund within the Human Potential Operating Programme.

References

  1. 1.
    WHO. Leishmaniasis (2017) [cited 2017 2017-11-30]; Available from: http://www.who.int/leishmaniasis/en/
  2. 2.
    Alvar J et al (2012) Leishmaniasis worldwide and global estimates of its incidence. PLoS One 7:e35671CrossRefGoogle Scholar
  3. 3.
    Murray HW et al (2005) Advances in leishmaniasis. Lancet 366:1561–1577CrossRefGoogle Scholar
  4. 4.
    Chappuis F et al (2007) Visceral leishmaniasis: what are the needs for diagnosis, treatment and control? Nat Rev Microbiol 5:873–882CrossRefGoogle Scholar
  5. 5.
    Fernández-Cotrina J et al (2013) Experimental model for reproduction of canine visceral leishmaniosis by Leishmania infantum. Vet Parasitol 192:118–128CrossRefGoogle Scholar
  6. 6.
    Melby PC et al (2001) The hamster as a model of human visceral Leishmaniasis: progressive disease and impaired generation of nitric oxide in the face of a prominent Th1-like cytokine response. J Immunol 166:1912–1920CrossRefGoogle Scholar
  7. 7.
    Aslan H et al (2013) A new model of progressive visceral leishmaniasis in hamsters by natural transmission via bites of vector sand flies. J Infect Dis 207:1328–1338CrossRefGoogle Scholar
  8. 8.
    Loeuillet C et al (2016) Study of Leishmania pathogenesis in mice: experimental considerations. Parasit Vectors 9:144CrossRefGoogle Scholar
  9. 9.
    Scott P, Novais FO (2016) Cutaneous leishmaniasis: immune responses in protection and pathogenesis. Nat Rev Immunol 16:581–592CrossRefGoogle Scholar
  10. 10.
    Leclercq V et al (1996) The outcome of the parasitic process initiated by Leishmania infantum in laboratory mice: a tissue-dependent pattern controlled by the Lsh and MHC loci. J Immunol 157:4537–4545PubMedGoogle Scholar
  11. 11.
    Kaye PM, Beattie L (2016) Lessons from other diseases: granulomatous inflammation in leishmaniasis. Semin Immunopathol 38:249–260CrossRefGoogle Scholar
  12. 12.
    Atayde VD (2015) Exosome secretion by the parasitic protozoan Leishmania within the sand fly midgut. Cell Rep 13:957–967CrossRefGoogle Scholar
  13. 13.
    Gomes R, Oliveira F (2012) The immune response to sand fly salivary proteins and its influence on Leishmania immunity. Front Immunol 3:110CrossRefGoogle Scholar
  14. 14.
    Rogers ME (2012) The role of Leishmania proteophosphoglycans in sand fly transmission and infection of the mammalian host. Front Microbiol 3:223CrossRefGoogle Scholar
  15. 15.
    Dey R et al (2018) Gut microbes egested during bites of infected sand flies augment severity of leishmaniasis via inflammasome-derived IL-1β. Cell Host Microbe 23:134–143CrossRefGoogle Scholar
  16. 16.
    Sacks DL, Melby PC (2015) Animal models for the analysis of immune responses to leishmaniasis. Curr Protoc Immunol 108:19.2.1–19.2.24CrossRefGoogle Scholar
  17. 17.
    Faria J et al (2016) Leishmania infantum asparagine synthetase a is dispensable for parasites survival and infectivity. PLoS Negl Trop Dis 10:e0004365CrossRefGoogle Scholar
  18. 18.
    Faria J et al (2016) Disclosing the essentiality of ribose-5-phosphate isomerase B in Trypanosomatids. Sci Rep 6:26937CrossRefGoogle Scholar
  19. 19.
    Costa Lima SA et al (2012) Characterization and evaluation of BNIPDaoct-loaded PLGA nanoparticles for visceral leishmaniasis: in vitro and in vivo studies. Nanomedicine (Lond) 7:1839–1849CrossRefGoogle Scholar
  20. 20.
    Tavares J et al (2012) Anti-leishmanial activity of the bisnaphthalimidopropyl derivatives. Parasitol Int 61:360–363CrossRefGoogle Scholar
  21. 21.
    Pérez-Cabezas B et al (2016) Interleukin-27 early impacts Leishmania infantum infection in mice and correlates with active visceral disease in humans. Front Immunol 7:478CrossRefGoogle Scholar
  22. 22.
    Nascimento MS et al (2016) NOD2-RIP2-mediated signaling helps shape adaptive immunity in visceral leishmaniasis. J Infect Dis 214:1647–1657CrossRefGoogle Scholar
  23. 23.
    Arcanjo AF et al (2017) Toll-like receptor 2 is required for inflammatory process development during Leishmania infantum infection. Front Microbiol 8:978CrossRefGoogle Scholar
  24. 24.
    Silvestre R et al (2007) SIR2-deficient Leishmania infantum induces a defined IFN-gamma/IL-10 pattern that correlates with protection. J Immunol 179:3161–3170CrossRefGoogle Scholar
  25. 25.
    Agallou M et al (2017) Identification of BALB/c immune markers correlated with a partial protection to Leishmania infantum after vaccination with a rationally designed multi-epitope cysteine protease A peptide-based Nanovaccine. PLoS Negl Trop Dis 11:e0005311CrossRefGoogle Scholar
  26. 26.
    Banerjee A et al (2018) Live attenuated Leishmania donovani centrin gene-deleted parasites induce IL-23-dependent IL-17-protective immune response against visceral leishmaniasis in a murine model. J Immunol 200:163–176CrossRefGoogle Scholar
  27. 27.
    Buffet PA et al (1995) Culture microtitration: a sensitive method for quantifying Leishmania infantum in tissues of infected mice. Antimicrob Agents Chemother 39:2167–2168CrossRefGoogle Scholar
  28. 28.
    Nicolas L et al (2002) Real-time PCR for detection and quantitation of Leishmania in mouse tissues. J Clin Microbiol 40:1666–1669CrossRefGoogle Scholar
  29. 29.
    Cunha J et al (2013) Characterization of the biology and infectivity of Leishmania infantum viscerotropic and dermotropic strains isolated from HIV+ and HIV- patients in the murine model of visceral leishmaniasis. Parasit Vectors 6:122CrossRefGoogle Scholar
  30. 30.
    Moreira D et al (2015) Leishmania infantum modulates host macrophage mitochondrial metabolism by hijacking the SIRT1-AMPK axis. PLoS Pathog 11:e1004684CrossRefGoogle Scholar
  31. 31.
    Michel G et al (2011) Luciferase-expressing Leishmania infantum allows the monitoring of amastigote population size, in vivo, ex vivo and in vitro. PLoS Negl Trop Dis 5:e1323CrossRefGoogle Scholar
  32. 32.
    Reimao JQ et al (2015) Generation of luciferase-expressing Leishmania infantum chagasi and assessment of miltefosine efficacy in infected hamsters through bioimaging. PLoS Negl Trop Dis 9:e0003556CrossRefGoogle Scholar
  33. 33.
    Melo GD et al (2017) New insights into experimental visceral leishmaniasis: real-time in vivo imaging of Leishmania donovani virulence. PLoS Negl Trop Dis 11:e0005924CrossRefGoogle Scholar
  34. 34.
    Costa DM et al (2018) Whole-mouse in vivo bioluminescence imaging applied to drug screening against Leishmania infantum: a reliable method to evaluate efficacy and optimize treatment regimens. bioRxiv.  https://doi.org/10.1101/326355
  35. 35.
    Sereno D et al (2001) DNA transformation of Leishmania infantum axenic amastigotes and their use in drug screening. Antimicrob Agents Chemother 45:1168–1173CrossRefGoogle Scholar
  36. 36.
    Tavares J et al (2017) In vivo imaging of pathogen homing to the host tissues. Methods 127:37–44CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Joana Tavares
    • 1
    • 2
  • Nuno Santarém
    • 1
    • 2
  • Anabela Cordeiro-da-Silva
    • 1
    • 2
    • 3
    Email author
  1. 1.i3S-Instituto de Investigação e Inovação em SaúdeUniversidade do PortoPortoPortugal
  2. 2.IBMC–Instituto de Biologia Molecular e Celular, Parasite Disease GroupUniversidade do PortoPortoPortugal
  3. 3.Departamento de Ciências Biológicas, Faculdade de FarmáciaUniversidade do PortoPortoPortugal

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