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...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | http://europepmc.org/articles/PMC3265455?pdf=render |
| _version_ | 1818192909505134592 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-12T00:38:00Z |
| format | Article |
| id | doaj.art-98e4fabfce13432bbbb55a03dd8346fa |
| institution | Directory Open Access Journal |
| issn | 1935-2727 1935-2735 |
| language | English |
| last_indexed | 2024-12-12T00:38:00Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-98e4fabfce13432bbbb55a03dd8346fa2022-12-22T00:44:19ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352012-01-0161e146910.1371/journal.pntd.0001469Impact of continuous axenic cultivation in Leishmania infantum virulence.Diana MoreiraNuno SantarémInês LoureiroJoana TavaresAna Marta SilvaAna Marina AmorimAli OuaissiAnabela Cordeiro-da-SilvaRicardo SilvestreExperimental 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.http://europepmc.org/articles/PMC3265455?pdf=render |
| spellingShingle | 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. PLoS Neglected Tropical Diseases |
| title | 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_short | Impact of continuous axenic cultivation in Leishmania infantum virulence. |
| title_sort | impact of continuous axenic cultivation in leishmania infantum virulence |
| url | http://europepmc.org/articles/PMC3265455?pdf=render |
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