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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 E 2  (PGE 2 ), 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 PGE 2  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 E 2  (PGE 2 ) 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 PGE 2  and COX-2 might contribute to the establishment and maintenance of chronic phase of the  T. gondii  infection in muscle cell.

parasites and vectors

Toxoplasma gondii- skeletal muscle cells interaction increases lipid droplet biogenesis and positively modulates the production of IL-12, IFN-g and PGE2