Toxoplasma gondii- skeletal muscle cells interaction increases lipid droplet biogenesis and positively modulates the production of IL-12, IFN-g and PGE2
Abstract Background The interest in the mechanisms involved in Toxoplasma gondii lipid acquisition has steadily increased during the past few decades, but it remains not completely understood. Here, we investigated the biogenesis and the fate of lipid droplets (LD) of skeletal muscle cells (SkMC) du...
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| Format: | Article |
| Language: | English |
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BMC
2014-01-01
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| Series: | Parasites & Vectors |
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| Online Access: | https://doi.org/10.1186/1756-3305-7-47 |
| _version_ | 1797811779539042304 |
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| author | Alessandra F Gomes Kelly G Magalhães Renata M Rodrigues Laís de Carvalho Raphael Molinaro Patrícia T Bozza Helene S Barbosa |
| author_facet | Alessandra F Gomes Kelly G Magalhães Renata M Rodrigues Laís de Carvalho Raphael Molinaro Patrícia T Bozza Helene S Barbosa |
| author_sort | Alessandra F Gomes |
| collection | DOAJ |
| description | Abstract Background The interest in the mechanisms involved in Toxoplasma gondii lipid acquisition has steadily increased during the past few decades, but it remains not completely understood. Here, we investigated the biogenesis and the fate of lipid droplets (LD) of skeletal muscle cells (SkMC) during their interaction with T. gondii by confocal and electron microscopy. We also evaluated whether infected SkMC modulates the production of prostaglandin E2 (PGE2), cytokines interleukin-12 (IL-12) and interferon-gamma (INF-g), and also the cyclooxygenase-2 (COX-2) gene induction. Methods Primary culture of skeletal muscle cells were infected with tachyzoites of T. gondii and analysed by confocal microscopy for observation of LD. Ultrastructural cytochemistry was also used for lipid and sarcoplasmatic reticulum (SR) detection. Dosage of cytokines (IL-12 and INF-g) by ELISA technique and enzyme-linked immunoassay (EIA) for PGE2 measurement were employed. The COX-2 gene expression analysis was performed by real time reverse transcriptase polymerase chain reaction (qRT-PCR). Results We demonstrated that T. gondii infection of SkMC leads to increase in LD number and area in a time course dependent manner. Moreover, the ultrastructural analysis demonstrated that SR and LD are in direct contact with parasitophorous vacuole membrane (PVM), within the vacuolar matrix, around it and interacting directly with the membrane of parasite, indicating that LD are recruited and deliver their content inside the parasitophorous vacuole (PV) in T. gondii-infected SkMC. We also observed a positive modulation of the production of IL-12 and IFN-g, increase of COX-2 mRNA levels in the first hour of T. gondii-SkMC interaction and an increase of prostaglandin E2 (PGE2) synthesis from 6 h up to 48 h of infection. Conclusions Taken together, the close association between SR and LD with PV could represent a source of lipids as well as other nutrients for the parasite survival, and together with the increased levels of IL-12, INF-g and inflammatory indicators PGE2 and COX-2 might contribute to the establishment and maintenance of chronic phase of the T. gondii infection in muscle cell. |
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| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-03-13T07:28:47Z |
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| spelling | doaj.art-c46e6eabe63b4e52885839a8f8345ccf2023-06-04T11:15:34ZengBMCParasites & Vectors1756-33052014-01-017111310.1186/1756-3305-7-47Toxoplasma gondii- skeletal muscle cells interaction increases lipid droplet biogenesis and positively modulates the production of IL-12, IFN-g and PGE2Alessandra F Gomes0Kelly G Magalhães1Renata M Rodrigues2Laís de Carvalho3Raphael Molinaro4Patrícia T Bozza5Helene S Barbosa6Laboratório de Biologia Estrutural, Instituto Oswaldo Cruz, Fundação Oswaldo CruzLaboratório de Imunologia e Inflamação, Universidade de BrasíliaLaboratório de Biologia Estrutural, Instituto Oswaldo Cruz, Fundação Oswaldo CruzLaboratório Cultura de Células, Depto. Histologia e Embriologia, Instituto de Biologia, Universidade do Estado do Rio de JaneiroLaboratório de Imunofarmacologia, Instituto Oswaldo Cruz, Fundação Oswaldo CruzLaboratório de Imunofarmacologia, Instituto Oswaldo Cruz, Fundação Oswaldo CruzLaboratório de Biologia Estrutural, Instituto Oswaldo Cruz, Fundação Oswaldo CruzAbstract Background The interest in the mechanisms involved in Toxoplasma gondii lipid acquisition has steadily increased during the past few decades, but it remains not completely understood. Here, we investigated the biogenesis and the fate of lipid droplets (LD) of skeletal muscle cells (SkMC) during their interaction with T. gondii by confocal and electron microscopy. We also evaluated whether infected SkMC modulates the production of prostaglandin E2 (PGE2), cytokines interleukin-12 (IL-12) and interferon-gamma (INF-g), and also the cyclooxygenase-2 (COX-2) gene induction. Methods Primary culture of skeletal muscle cells were infected with tachyzoites of T. gondii and analysed by confocal microscopy for observation of LD. Ultrastructural cytochemistry was also used for lipid and sarcoplasmatic reticulum (SR) detection. Dosage of cytokines (IL-12 and INF-g) by ELISA technique and enzyme-linked immunoassay (EIA) for PGE2 measurement were employed. The COX-2 gene expression analysis was performed by real time reverse transcriptase polymerase chain reaction (qRT-PCR). Results We demonstrated that T. gondii infection of SkMC leads to increase in LD number and area in a time course dependent manner. Moreover, the ultrastructural analysis demonstrated that SR and LD are in direct contact with parasitophorous vacuole membrane (PVM), within the vacuolar matrix, around it and interacting directly with the membrane of parasite, indicating that LD are recruited and deliver their content inside the parasitophorous vacuole (PV) in T. gondii-infected SkMC. We also observed a positive modulation of the production of IL-12 and IFN-g, increase of COX-2 mRNA levels in the first hour of T. gondii-SkMC interaction and an increase of prostaglandin E2 (PGE2) synthesis from 6 h up to 48 h of infection. Conclusions Taken together, the close association between SR and LD with PV could represent a source of lipids as well as other nutrients for the parasite survival, and together with the increased levels of IL-12, INF-g and inflammatory indicators PGE2 and COX-2 might contribute to the establishment and maintenance of chronic phase of the T. gondii infection in muscle cell.https://doi.org/10.1186/1756-3305-7-47Toxoplasma gondiiLipid dropletsSkeletal muscle cellsProstaglandin-E2Cytokines |
| spellingShingle | Alessandra F Gomes Kelly G Magalhães Renata M Rodrigues Laís de Carvalho Raphael Molinaro Patrícia T Bozza Helene S Barbosa Toxoplasma gondii- skeletal muscle cells interaction increases lipid droplet biogenesis and positively modulates the production of IL-12, IFN-g and PGE2 Parasites & Vectors Toxoplasma gondii Lipid droplets Skeletal muscle cells Prostaglandin-E2 Cytokines |
| title | Toxoplasma gondii- skeletal muscle cells interaction increases lipid droplet biogenesis and positively modulates the production of IL-12, IFN-g and PGE2 |
| title_full | Toxoplasma gondii- skeletal muscle cells interaction increases lipid droplet biogenesis and positively modulates the production of IL-12, IFN-g and PGE2 |
| title_fullStr | Toxoplasma gondii- skeletal muscle cells interaction increases lipid droplet biogenesis and positively modulates the production of IL-12, IFN-g and PGE2 |
| title_full_unstemmed | Toxoplasma gondii- skeletal muscle cells interaction increases lipid droplet biogenesis and positively modulates the production of IL-12, IFN-g and PGE2 |
| title_short | Toxoplasma gondii- skeletal muscle cells interaction increases lipid droplet biogenesis and positively modulates the production of IL-12, IFN-g and PGE2 |
| title_sort | toxoplasma gondii skeletal muscle cells interaction increases lipid droplet biogenesis and positively modulates the production of il 12 ifn g and pge2 |
| topic | Toxoplasma gondii Lipid droplets Skeletal muscle cells Prostaglandin-E2 Cytokines |
| url | https://doi.org/10.1186/1756-3305-7-47 |
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