Impact of continuous axenic cultivation in Leishmania infantum virulence.
Experimental infections with visceral Leishmania spp. are frequently performed referring to stationary parasite cultures that are comprised of a mixture of metacyclic and non-metacyclic parasites often with little regard to time of culture and metacyclic purification. This may lead to misleading or...
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ftdoajarticles:oai:doaj.org/article:98e4fabfce13432bbbb55a03dd8346fa 2023-05-15T15:12:44+02:00 Impact of continuous axenic cultivation in Leishmania infantum virulence. Diana Moreira Nuno Santarém Inês Loureiro Joana Tavares Ana Marta Silva Ana Marina Amorim Ali Ouaissi Anabela Cordeiro-da-Silva Ricardo Silvestre 2012-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0001469 https://doaj.org/article/98e4fabfce13432bbbb55a03dd8346fa EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3265455?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 doi:10.1371/journal.pntd.0001469 1935-2727 1935-2735 https://doaj.org/article/98e4fabfce13432bbbb55a03dd8346fa PLoS Neglected Tropical Diseases, Vol 6, Iss 1, p e1469 (2012) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2012 ftdoajarticles https://doi.org/10.1371/journal.pntd.0001469 2022-12-31T02:23:35Z Experimental infections with visceral Leishmania spp. are frequently performed referring to stationary parasite cultures that are comprised of a mixture of metacyclic and non-metacyclic parasites often with little regard to time of culture and metacyclic purification. This may lead to misleading or irreproducible experimental data. It is known that the maintenance of Leishmania spp. in vitro results in a progressive loss of virulence that can be reverted by passage in a mammalian host. In the present study, we aimed to characterize the loss of virulence in culture comparing the in vitro and in vivo infection and immunological profile of L. infantum stationary promastigotes submitted to successive periods of in vitro cultivation. To evaluate the effect of axenic in vitro culture in parasite virulence, we submitted L. infantum promastigotes to 4, 21 or 31 successive in vitro passages. Our results demonstrated a rapid and significant loss of parasite virulence when parasites are sustained in axenic culture. Strikingly, the parasite capacity to modulate macrophage activation decreased significantly with the augmentation of the number of in vitro passages. We validated these in vitro observations using an experimental murine model of infection. A significant correlation was found between higher parasite burdens and lower number of in vitro passages in infected Balb/c mice. Furthermore, we have demonstrated that the virulence deficit caused by successive in vitro passages results from an inadequate capacity to differentiate into amastigote forms. In conclusion, our data demonstrated that the use of parasites with distinct periods of axenic in vitro culture induce distinct infection rates and immunological responses and correlated this phenotype with a rapid loss of promastigote differentiation capacity. These results highlight the need for a standard operating protocol (SOP) when studying Leishmania species. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 6 1 e1469 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Diana Moreira Nuno Santarém Inês Loureiro Joana Tavares Ana Marta Silva Ana Marina Amorim Ali Ouaissi Anabela Cordeiro-da-Silva Ricardo Silvestre Impact of continuous axenic cultivation in Leishmania infantum virulence. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Experimental infections with visceral Leishmania spp. are frequently performed referring to stationary parasite cultures that are comprised of a mixture of metacyclic and non-metacyclic parasites often with little regard to time of culture and metacyclic purification. This may lead to misleading or irreproducible experimental data. It is known that the maintenance of Leishmania spp. in vitro results in a progressive loss of virulence that can be reverted by passage in a mammalian host. In the present study, we aimed to characterize the loss of virulence in culture comparing the in vitro and in vivo infection and immunological profile of L. infantum stationary promastigotes submitted to successive periods of in vitro cultivation. To evaluate the effect of axenic in vitro culture in parasite virulence, we submitted L. infantum promastigotes to 4, 21 or 31 successive in vitro passages. Our results demonstrated a rapid and significant loss of parasite virulence when parasites are sustained in axenic culture. Strikingly, the parasite capacity to modulate macrophage activation decreased significantly with the augmentation of the number of in vitro passages. We validated these in vitro observations using an experimental murine model of infection. A significant correlation was found between higher parasite burdens and lower number of in vitro passages in infected Balb/c mice. Furthermore, we have demonstrated that the virulence deficit caused by successive in vitro passages results from an inadequate capacity to differentiate into amastigote forms. In conclusion, our data demonstrated that the use of parasites with distinct periods of axenic in vitro culture induce distinct infection rates and immunological responses and correlated this phenotype with a rapid loss of promastigote differentiation capacity. These results highlight the need for a standard operating protocol (SOP) when studying Leishmania species. |
format |
Article in Journal/Newspaper |
author |
Diana Moreira Nuno Santarém Inês Loureiro Joana Tavares Ana Marta Silva Ana Marina Amorim Ali Ouaissi Anabela Cordeiro-da-Silva Ricardo Silvestre |
author_facet |
Diana Moreira Nuno Santarém Inês Loureiro Joana Tavares Ana Marta Silva Ana Marina Amorim Ali Ouaissi Anabela Cordeiro-da-Silva Ricardo Silvestre |
author_sort |
Diana Moreira |
title |
Impact of continuous axenic cultivation in Leishmania infantum virulence. |
title_short |
Impact of continuous axenic cultivation in Leishmania infantum virulence. |
title_full |
Impact of continuous axenic cultivation in Leishmania infantum virulence. |
title_fullStr |
Impact of continuous axenic cultivation in Leishmania infantum virulence. |
title_full_unstemmed |
Impact of continuous axenic cultivation in Leishmania infantum virulence. |
title_sort |
impact of continuous axenic cultivation in leishmania infantum virulence. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2012 |
url |
https://doi.org/10.1371/journal.pntd.0001469 https://doaj.org/article/98e4fabfce13432bbbb55a03dd8346fa |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 6, Iss 1, p e1469 (2012) |
op_relation |
http://europepmc.org/articles/PMC3265455?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 doi:10.1371/journal.pntd.0001469 1935-2727 1935-2735 https://doaj.org/article/98e4fabfce13432bbbb55a03dd8346fa |
op_doi |
https://doi.org/10.1371/journal.pntd.0001469 |
container_title |
PLoS Neglected Tropical Diseases |
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6 |
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1 |
container_start_page |
e1469 |
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