Letter | Published:

Sequential blood meals promote Leishmania replication and reverse metacyclogenesis augmenting vector infectivity

Nature Microbiologyvolume 3pages548555 (2018) | Download Citation

Abstract

Sand flies, similar to most vectors, take multiple blood meals during their lifetime1,2,3,4. The effect of subsequent blood meals on pathogens developing in the vector and their impact on disease transmission have never been examined. Here, we show that ingestion of a second uninfected blood meal by Leishmania-infected sand flies triggers dedifferentiation of metacyclic promastigotes, considered a terminally differentiated stage inside the vector5, to a leptomonad-like stage, the retroleptomonad promastigote. Reverse metacyclogenesis occurs after every subsequent blood meal where retroleptomonad promastigotes rapidly multiply and differentiate to metacyclic promastigotes enhancing sand fly infectiousness. Importantly, a subsequent blood meal amplifies the few Leishmania parasites acquired by feeding on infected hosts by 125-fold, and increases lesion frequency by fourfold, in twice-fed compared with single-fed flies. These findings place readily available blood sources as a critical element in transmission and propagation of vector-borne pathogens.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

  1. 1.

    Guzman, H., Walters, L. L. & Tesh, R. B. Histologic detection of multiple blood meals in Phlebotomus duboscqi (Diptera: Psychodidae). J. Med. Entomol. 31, 890–897 (1994).

  2. 2.

    Norris, L. C., Fornadel, C. M., Hung, W. C., Pineda, F. J. & Norris, D. E. Frequency of multiple blood meals taken in a single gonotrophic cycle by Anopheles arabiensis mosquitoes in Macha, Zambia. Am. Trop. Med. Hyg. 83, 33–37 (2010).

  3. 3.

    Kramer, L. D. & Ebel, G. D. Dynamics of flavivirus infection in mosquitoes. Adv. Virus Res. 60, 187–232 (2003).

  4. 4.

    Abbasi, I., Cunio, R. & Warburg, A. Identification of blood meals imbibed by phlebotomine sand flies using cytochrome b PCR and reverse line blotting. Vector Borne Zoonotic Dis. 9, 79–86 (2009).

  5. 5.

    Bates, P. A. Transmission of Leishmania metacyclic promastigotes by phlebotomine sand flies. Int. J. Parasitol. 37, 1097–1106 (2007).

  6. 6.

    Vector-Borne Diseases (WHO, 2014); http://apps.who.int/iris/handle/10665/206531?mode=full

  7. 7.

    Das, S., Muleba, M., Stevenson, J. C., Pringle, J. C. & Norris, D. E. Beyond the entomological inoculation rate: characterizing multiple blood feeding behavior and Plasmodium falciparum multiplicity of infection in Anopheles mosquitoes in northern Zambia. Parasit. Vectors 10, 45 (2017).

  8. 8.

    Dostalova, A. & Volf, P. Leishmania development in sand flies: parasite-vector interactions overview. Parasit. Vectors 5, 276 (2012).

  9. 9.

    Aslan, H. et al. A new model of progressive visceral leishmaniasis in hamsters by natural transmission via bites of vector sand flies. J. Infect. Dis. 207, 1328–1338 (2013).

  10. 10.

    Collin, N. et al. Sand fly salivary proteins induce strong cellular immunity in a natural reservoir of visceral leishmaniasis with adverse consequences for Leishmania. PLoS Pathog. 5, e1000441 (2009).

  11. 11.

    Howard, M. K., Sayers, G. & Miles, M. A. Leishmania donovani metacyclic promastigotes: transformation in vitro, lectin agglutination, complement resistance, and infectivity. Exp. Parasitol. 64, 147–156 (1987).

  12. 12.

    Serafim, T. D. et al. Leishmania metacyclogenesis is promoted in the absence of purines. PLoS Negl. Trop. Dis. 6, e1833 (2012).

  13. 13.

    Ready, P. D. Biology of phlebotomine sand flies as vectors of disease agents. Annu. Rev. Entomol. 58, 227–250 (2013).

  14. 14.

    Kamhawi, S. Phlebotomine sand flies and Leishmania parasites: friends or foes? Trends Parasitol. 22, 439–445 (2006).

  15. 15.

    Alexander, B., de Carvalho, R. L., McCallum, H. & Pereira, M. H. Role of the domestic chicken (Gallus gallus) in the epidemiology of urban visceral leishmaniasis in Brazil. Emerg. Infect. Dis. 8, 1480–1485 (2002).

  16. 16.

    Sant’anna, M. R. et al. Chicken blood provides a suitable meal for the sand fly Lutzomyia longipalpis and does not inhibit Leishmania development in the gut. Parasit. Vectors 3, 3 (2010).

  17. 17.

    Guimaraes, E. S. A. S. et al Leishmania infection and blood food sources of phlebotomines in an area of Brazil endemic for visceral and tegumentary leishmaniasis. PLoS ONE 12, e0179052 (2017).

  18. 18.

    Rogers, M. E. The role of Leishmania proteophosphoglycans in sand fly transmission and infection of the mammalian host. Front Microbiol. 3, 223 (2012).

  19. 19.

    Kimblin, N. et al. Quantification of the infectious dose of Leishmania major transmitted to the skin by single sand flies. Proc. Natl Acad. Sci. USA 105, 10125–10130 (2008).

  20. 20.

    Bates, P. A. Leishmania sand fly interaction: progress and challenges. Curr. Opin. Microbiol. 11, 340–344 (2008).

  21. 21.

    Gossage, S. M., Rogers, M. E. & Bates, P. A. Two separate growth phases during the development of Leishmania in sand flies: implications for understanding the life cycle. Int. J. Parasitol. 33, 1027–1034 (2003).

  22. 22.

    Killick-Kendrick, R. & Rioux, J. A. Mark-release-recapture of sand flies fed on leishmanial dogs: the natural life-cycle of Leishmania infantum in Phlebotomus ariasi. Parassitologia 44, 67–71 (2002).

  23. 23.

    Lawyer, P., Killick-Kendrick, M., Rowland, T., Rowton, E. & Volf, P. Laboratory colonization and mass rearing of phlebotomine sand flies (Diptera, Psychodidae). Parasite 24, 42 (2017).

  24. 24.

    Oliveira, F. et al. A sand fly salivary protein vaccine shows efficacy against vector-transmitted cutaneous leishmaniasis in nonhuman primates. Sci. Transl. Med. 7, 290ra290 (2015).

  25. 25.

    Sacks, D. L. & Melby, P. C. Animal models for the analysis of immune responses to leishmaniasis. Curr. Protoc. Immunol. 108, 11–24 (2015).

  26. 26.

    Sacks, D. L. & Perkins, P. V. Identification of an infective stage of Leishmania promastigotes. Science 223, 1417–1419 (1984).

  27. 27.

    Gomes, R. et al. Immunity to a salivary protein of a sand fly vector protects against the fatal outcome of visceral leishmaniasis in a hamster model. Proc. Natl Acad. Sci. USA 105, 7845–7850 (2008).

  28. 28.

    Selvapandiyan, A. et al. Intracellular replication-deficient Leishmania donovani induces long lasting protective immunity against visceral leishmaniasis. J. Immunol. 183, 1813–1820 (2009).

  29. 29.

    Kamhawi, S., Belkaid, Y., Modi, G., Rowton, E. & Sacks, D. Protection against cutaneous leishmaniasis resulting from bites of uninfected sand flies. Science 290, 1351–1354 (2000).

  30. 30.

    Schneider, C. A., Rasband, W. S. & Eliceiri, K. W. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9, 671–675 (2012).

Download references

Acknowledgements

We would like to thank E. Fischer and S. Ricklefs from the Research Technology Branch (RTB), NIAID, for electron microscopy support; R. Kissinger from RTB, NIAID, for illustration support; A. Perkins and W. de Castro from VMBS, NIAID, for technical support; V. Vernyuy, T.R. Wilson and B.G. Bonilla from LMVR, NIAID for sand fly insectary support; R. Dey and H. Nakhasi from CBER, FDA, for help with qPCR; A.M.A. Souza for help with statistical analysis and C. Barillas-Mury and J.M.C. Ribeiro from LMVR, NIAID, for critical reading of the manuscript. This research was supported by the Intramural Research Program of the NIH, National Institute of Allergy and Infectious Diseases.

Author information

Affiliations

  1. Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD, USA

    • Tiago D. Serafim
    • , Iliano V. Coutinho-Abreu
    • , Fabiano Oliveira
    • , Claudio Meneses
    • , Shaden Kamhawi
    •  & Jesus G. Valenzuela

Authors

  1. Search for Tiago D. Serafim in:

  2. Search for Iliano V. Coutinho-Abreu in:

  3. Search for Fabiano Oliveira in:

  4. Search for Claudio Meneses in:

  5. Search for Shaden Kamhawi in:

  6. Search for Jesus G. Valenzuela in:

Contributions

T.D.S. and I.V.C.A. designed and performed the experiments. T.D.S. analysed the data. I.V.C.A analysed qPCR data. C.M. performed sand fly insectary work. J.G.V., S.K. and F.O. were involved in the design, interpretation and supervision of this study. All authors wrote the manuscript.

Competing interests

The authors declare no competing interests

Corresponding authors

Correspondence to Shaden Kamhawi or Jesus G. Valenzuela.

Supplementary information

  1. Supplementary Information

    Supplementary Figures 1–4. Supplementary Tables 1–3. Legends for Supplementary Videos 1–8.

  2. Life Sciences Reporting Summary(PDF 64 kb)

Videos

  1. Supplementary Video 1

    Fast-swimming metacyclics at mature infection.

  2. Supplementary Video 2

    Fast-swimming metacyclics at mature infection.

  3. Supplementary Video 3

    Fast-swimming metacyclics dedifferentiate into slow moving retroleptomonads after a subsequent uninfected blood meal.

  4. Supplementary Video 4

    Fast-swimming metacyclics dedifferentiate into slow moving retroleptomonads after a subsequent uninfected blood meal.

  5. Supplementary Video 5

    Dedifferentiation of a metacyclic promastigote – representative event 1.

  6. Supplementary Video 6

    Dedifferentiation of a metacyclic promastigote – representativeevent 2.

  7. Supplementary Video 7

    Dedifferentiation of a metacyclic promastigote – representativeevent 3.

  8. Supplementary Video 8

    The haptomonad parasite sphere of an infected sand fly after a subsequent uninfected blood meal.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/s41564-018-0125-7

Further reading