Wolbachia introduction into Lutzomyia longipalpis (Diptera: Psychodidae) cell lines and its effects on immune-related gene expression and interaction with Leishmania infantum

Daniela Da Silva Gonçalves, Iñaki Iturbe-Ormaetxe, Andrea Martins-Da-Silva, Erich Loza Telleria, Marcele Neves Rocha, Yara M. Traub-Csekö, Scott L. O'Neill, Maurício Roberto Viana Sant'Anna, Luciano Andrade Moreira

Research output: Contribution to journalArticleResearchpeer-review

2 Citations (Scopus)

Abstract

Background: The leishmaniases are important neglected diseases caused by Leishmania spp. which are transmitted by sand flies, Lutzomyia longipalpis being the main vector of visceral leishmaniasis in the Americas. The methodologies for leishmaniasis control are not efficient, causing 1.5 million reported cases annually worldwide, therefore showing the need for development of novel strategies and interventions to control transmission of the disease. The bacterium Wolbachia pipientis is being used to control viruses transmitted by mosquitoes, such as dengue and Zika, and its introduction in disease vectors has been effective against parasites such as Plasmodium. Here we show the first successful establishment of Wolbachia into two different embryonic cell lines from L. longipalpis, LL-5 and Lulo, and analysed its effects on the sand fly innate immune system, followed by in vitro Leishmania infantum interaction. Results: Our results show that LL-5 cells respond to wMel and wMelPop-CLA strains within the first 72 h post-infection, through the expression of antimicrobial peptides and inducible nitric oxide synthase resulting in a decrease of Wolbachia detection in the early stages of infection. In subsequent passages, the wMel strain was not able to infect any of the sand fly cell lines while the wMelPop-CLA strain was able to stably infect Lulo cells and LL-5 at lower levels. In Wolbachia stably infected cells, the expression of immune-related genes involved with downregulation of the IMD, Toll and Jak-Stat innate immune pathways was significantly decreased, in comparison with the uninfected control, suggesting immune activation upon Wolbachia transinfection. Furthermore, Wolbachia transinfection did not promote a negative effect on parasite load in those cells. Conclusions: Initial strong immune responses of LL5 cells might explain the inefficiency of stable infections in these cells while we found that Lulo cells are more permissive to infection with Wolbachia causing an effect on the cell immune system, but not against in vitro L. infantum interaction. This establishes Lulo cells as a good system for the adaptation of Wolbachia in L. longipalpis.

Original languageEnglish
Article number33
Number of pages13
JournalParasites and Vectors
Volume12
Issue number1
DOIs
Publication statusPublished - 15 Jan 2019

Keywords

  • Leishmania infantum
  • LL-5 cell
  • Lulo cell
  • Lutzomyia longipalpis
  • Wolbachia

Cite this

Da Silva Gonçalves, D., Iturbe-Ormaetxe, I., Martins-Da-Silva, A., Telleria, E. L., Rocha, M. N., Traub-Csekö, Y. M., ... Moreira, L. A. (2019). Wolbachia introduction into Lutzomyia longipalpis (Diptera: Psychodidae) cell lines and its effects on immune-related gene expression and interaction with Leishmania infantum. Parasites and Vectors, 12(1), [33]. https://doi.org/10.1186/s13071-018-3227-4
Da Silva Gonçalves, Daniela ; Iturbe-Ormaetxe, Iñaki ; Martins-Da-Silva, Andrea ; Telleria, Erich Loza ; Rocha, Marcele Neves ; Traub-Csekö, Yara M. ; O'Neill, Scott L. ; Sant'Anna, Maurício Roberto Viana ; Moreira, Luciano Andrade. / Wolbachia introduction into Lutzomyia longipalpis (Diptera: Psychodidae) cell lines and its effects on immune-related gene expression and interaction with Leishmania infantum. In: Parasites and Vectors. 2019 ; Vol. 12, No. 1.
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title = "Wolbachia introduction into Lutzomyia longipalpis (Diptera: Psychodidae) cell lines and its effects on immune-related gene expression and interaction with Leishmania infantum",
abstract = "Background: The leishmaniases are important neglected diseases caused by Leishmania spp. which are transmitted by sand flies, Lutzomyia longipalpis being the main vector of visceral leishmaniasis in the Americas. The methodologies for leishmaniasis control are not efficient, causing 1.5 million reported cases annually worldwide, therefore showing the need for development of novel strategies and interventions to control transmission of the disease. The bacterium Wolbachia pipientis is being used to control viruses transmitted by mosquitoes, such as dengue and Zika, and its introduction in disease vectors has been effective against parasites such as Plasmodium. Here we show the first successful establishment of Wolbachia into two different embryonic cell lines from L. longipalpis, LL-5 and Lulo, and analysed its effects on the sand fly innate immune system, followed by in vitro Leishmania infantum interaction. Results: Our results show that LL-5 cells respond to wMel and wMelPop-CLA strains within the first 72 h post-infection, through the expression of antimicrobial peptides and inducible nitric oxide synthase resulting in a decrease of Wolbachia detection in the early stages of infection. In subsequent passages, the wMel strain was not able to infect any of the sand fly cell lines while the wMelPop-CLA strain was able to stably infect Lulo cells and LL-5 at lower levels. In Wolbachia stably infected cells, the expression of immune-related genes involved with downregulation of the IMD, Toll and Jak-Stat innate immune pathways was significantly decreased, in comparison with the uninfected control, suggesting immune activation upon Wolbachia transinfection. Furthermore, Wolbachia transinfection did not promote a negative effect on parasite load in those cells. Conclusions: Initial strong immune responses of LL5 cells might explain the inefficiency of stable infections in these cells while we found that Lulo cells are more permissive to infection with Wolbachia causing an effect on the cell immune system, but not against in vitro L. infantum interaction. This establishes Lulo cells as a good system for the adaptation of Wolbachia in L. longipalpis.",
keywords = "Leishmania infantum, LL-5 cell, Lulo cell, Lutzomyia longipalpis, Wolbachia",
author = "{Da Silva Gon{\cc}alves}, Daniela and I{\~n}aki Iturbe-Ormaetxe and Andrea Martins-Da-Silva and Telleria, {Erich Loza} and Rocha, {Marcele Neves} and Traub-Csek{\"o}, {Yara M.} and O'Neill, {Scott L.} and Sant'Anna, {Maur{\'i}cio Roberto Viana} and Moreira, {Luciano Andrade}",
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Da Silva Gonçalves, D, Iturbe-Ormaetxe, I, Martins-Da-Silva, A, Telleria, EL, Rocha, MN, Traub-Csekö, YM, O'Neill, SL, Sant'Anna, MRV & Moreira, LA 2019, 'Wolbachia introduction into Lutzomyia longipalpis (Diptera: Psychodidae) cell lines and its effects on immune-related gene expression and interaction with Leishmania infantum', Parasites and Vectors, vol. 12, no. 1, 33. https://doi.org/10.1186/s13071-018-3227-4

Wolbachia introduction into Lutzomyia longipalpis (Diptera: Psychodidae) cell lines and its effects on immune-related gene expression and interaction with Leishmania infantum. / Da Silva Gonçalves, Daniela; Iturbe-Ormaetxe, Iñaki; Martins-Da-Silva, Andrea; Telleria, Erich Loza; Rocha, Marcele Neves; Traub-Csekö, Yara M.; O'Neill, Scott L.; Sant'Anna, Maurício Roberto Viana; Moreira, Luciano Andrade.

In: Parasites and Vectors, Vol. 12, No. 1, 33, 15.01.2019.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Wolbachia introduction into Lutzomyia longipalpis (Diptera: Psychodidae) cell lines and its effects on immune-related gene expression and interaction with Leishmania infantum

AU - Da Silva Gonçalves, Daniela

AU - Iturbe-Ormaetxe, Iñaki

AU - Martins-Da-Silva, Andrea

AU - Telleria, Erich Loza

AU - Rocha, Marcele Neves

AU - Traub-Csekö, Yara M.

AU - O'Neill, Scott L.

AU - Sant'Anna, Maurício Roberto Viana

AU - Moreira, Luciano Andrade

PY - 2019/1/15

Y1 - 2019/1/15

N2 - Background: The leishmaniases are important neglected diseases caused by Leishmania spp. which are transmitted by sand flies, Lutzomyia longipalpis being the main vector of visceral leishmaniasis in the Americas. The methodologies for leishmaniasis control are not efficient, causing 1.5 million reported cases annually worldwide, therefore showing the need for development of novel strategies and interventions to control transmission of the disease. The bacterium Wolbachia pipientis is being used to control viruses transmitted by mosquitoes, such as dengue and Zika, and its introduction in disease vectors has been effective against parasites such as Plasmodium. Here we show the first successful establishment of Wolbachia into two different embryonic cell lines from L. longipalpis, LL-5 and Lulo, and analysed its effects on the sand fly innate immune system, followed by in vitro Leishmania infantum interaction. Results: Our results show that LL-5 cells respond to wMel and wMelPop-CLA strains within the first 72 h post-infection, through the expression of antimicrobial peptides and inducible nitric oxide synthase resulting in a decrease of Wolbachia detection in the early stages of infection. In subsequent passages, the wMel strain was not able to infect any of the sand fly cell lines while the wMelPop-CLA strain was able to stably infect Lulo cells and LL-5 at lower levels. In Wolbachia stably infected cells, the expression of immune-related genes involved with downregulation of the IMD, Toll and Jak-Stat innate immune pathways was significantly decreased, in comparison with the uninfected control, suggesting immune activation upon Wolbachia transinfection. Furthermore, Wolbachia transinfection did not promote a negative effect on parasite load in those cells. Conclusions: Initial strong immune responses of LL5 cells might explain the inefficiency of stable infections in these cells while we found that Lulo cells are more permissive to infection with Wolbachia causing an effect on the cell immune system, but not against in vitro L. infantum interaction. This establishes Lulo cells as a good system for the adaptation of Wolbachia in L. longipalpis.

AB - Background: The leishmaniases are important neglected diseases caused by Leishmania spp. which are transmitted by sand flies, Lutzomyia longipalpis being the main vector of visceral leishmaniasis in the Americas. The methodologies for leishmaniasis control are not efficient, causing 1.5 million reported cases annually worldwide, therefore showing the need for development of novel strategies and interventions to control transmission of the disease. The bacterium Wolbachia pipientis is being used to control viruses transmitted by mosquitoes, such as dengue and Zika, and its introduction in disease vectors has been effective against parasites such as Plasmodium. Here we show the first successful establishment of Wolbachia into two different embryonic cell lines from L. longipalpis, LL-5 and Lulo, and analysed its effects on the sand fly innate immune system, followed by in vitro Leishmania infantum interaction. Results: Our results show that LL-5 cells respond to wMel and wMelPop-CLA strains within the first 72 h post-infection, through the expression of antimicrobial peptides and inducible nitric oxide synthase resulting in a decrease of Wolbachia detection in the early stages of infection. In subsequent passages, the wMel strain was not able to infect any of the sand fly cell lines while the wMelPop-CLA strain was able to stably infect Lulo cells and LL-5 at lower levels. In Wolbachia stably infected cells, the expression of immune-related genes involved with downregulation of the IMD, Toll and Jak-Stat innate immune pathways was significantly decreased, in comparison with the uninfected control, suggesting immune activation upon Wolbachia transinfection. Furthermore, Wolbachia transinfection did not promote a negative effect on parasite load in those cells. Conclusions: Initial strong immune responses of LL5 cells might explain the inefficiency of stable infections in these cells while we found that Lulo cells are more permissive to infection with Wolbachia causing an effect on the cell immune system, but not against in vitro L. infantum interaction. This establishes Lulo cells as a good system for the adaptation of Wolbachia in L. longipalpis.

KW - Leishmania infantum

KW - LL-5 cell

KW - Lulo cell

KW - Lutzomyia longipalpis

KW - Wolbachia

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