Azithromycin and spiramycin induce anti-inflammatory response in human trophoblastic (BeWo) cells infected by Toxoplasma gondii but are able to control infection

Abstract

Toxoplasma gondii is an important pathogen which may cause fetal infection if primary infection. Our previous studies have used human choriocarcinoma trophoblastic cells (BeWo cell line) as experimental model of T. gondii infection involving placental microenvironment. This study aimed to examine the effects of azithromycin and spiramycin against T. gondii infection in BeWo cells. Cells were treated with different concentrations of the macrolide antibiotics and analyzed first for cell viability using thiazolyl blue tetrazole (MTT) assay. As cell viability was significantly decreased with drug concentrations higher than 400 μg/mL, the concentration range used in further experiments was from 50 to 400 μg/mL. The number of infected cells and intracellular replication of T. gondii decreased after treatment with each drug. The infection induced up-regulation of the macrophage migration inhibitory factor (MIF), which was also enhanced in infected cells after treatment with azithromycin, but not with spiramycin. Analysis of the cytokine profile showed increase TNF-α, IL-10 and IL-4 production, but decreased IFN-γ levels, were detected in infected cells and treated with each drug. In conclusion, treatment of human trophoblastic BeWo cells with with azithromycin or spiramycin is able to control the infection and replication of T. gondii. In addition, treatment with these macrolides, especially with azityromycin induces an anti-inflammatory response and high MIF production, which can be important for the establishment and maintenance of a viable pregnancy during T. gondii infection.

Azithromycin is able to control Toxoplasma gondii infection in human villous explants

“…Both azithromycin and PSA were able to reduce T. gondii proliferation at both tested concentrations (200 and 1000 μg/mL). These data are in agreement with our previous studies, where azithromycin treatment was able to control T. gondii infection in human trophoblast BeWo cells [19]. Additionally, azithromycin also showed a protective role against the ME49 strain of T. gondii in rodent C. callosus , reducing vertical transmission of the parasite [18].…”

“…Given that azithromycin is considered safe during pregnancy and the fact that it may have activity against pathogens, such as Plasmodium and Toxoplasma , it has been suggested as a candidate for intermittent preventive treatment during pregnancy [7,17]. Previous studies have shown that azithromycin treatment in pregnant Calomys callosus and human trophoblast cells (lineage BeWo) was able to control T. gondii infection, suggesting that it may be an alternative drug for prevention of congenital infection [18,19]. Additionally, azithromycin has anti-inflammatory properties through modulating the production of proinflammatory cytokines that are produced during T. gondii infection [20].…”

IL10, TGF Beta1, and IFN Gamma Modulate Intracellular Signaling Pathways and Cytokine Production to Control Toxoplasma gondii Infection in BeWo Trophoblast Cells1

“…Samples were analyzed under BD flow cytometry (FACSCalibur; BD Company, San Diego, CA), and data were recorded using specialized BD Cell Quest software and CBA software. The results were expressed as mean fluorescence intensity (MFI) [17,19,41].…”

“…On the other hand, a normal pregnancy is characterized by a preferential Th2-type immune response and T regulatory cell responses [11], with the production of anti-inflammatory cytokines such as interleukin 10 (IL10) and transforming growth factor beta1 (TGFB1) by both maternal and fetal cells, including trophoblast cells [12,13]. The role of trophoblast cells in the immunology of pregnancy, especially in the presence of intracellular pathogens such as T. gondii and Neospora caninum, has been studied by our group using wellestablished cell lines, such as BeWo cells [14][15][16][17][18][19]. These cells were isolated from a human choriocarcinoma [20] and preserved their trophoblastic characteristics, such as hormone production and cytokine release under in vitro stimulation [21,22].…”

“…Even though BeWo cells are derived from a choriocarcinoma, they are becoming an excellent in vitro model to investigate T. gondii infection in trophoblast cells. In fact, our group has investigated several aspects of the immunology of pregnancy using these cells as a trophoblast model, including their cross-talk with monocytes [18] and the role of cytokines and drugs during T. gondii infection [14,15,17,19].…”