Abstract
Congenital toxoplasmosis is a serious health problem that can lead to miscarriage. HTR-8/SVneo is a first trimester extravillous trophoblast, while BeWo is a choriocarcinoma with properties of villous trophoblast cells. In the placenta, iron is taken up from Fe-transferrin through the transferrin receptor being the ion an important nutrient during pregnancy and also for Toxoplasma gondii proliferation. The aim of this study was to evaluate the role of iron in T. gondii proliferation in BeWo and HTR-8/SVneo cells and in human chorionic villous explants. The cells were infected with T. gondii, iron supplemented or deprived by holo-transferrin or deferoxamine, respectively, and parasite proliferation and genes related to iron balance were analyzed. It was verified that the addition of holo-transferrin increased, and DFO decreased the parasite multiplication in both trophoblastic cells, however, in a more expressive manner in HTR-8/SVneo, indicating that the parasite depends on iron storage in trophoblastic cells for its growth. Also, tachyzoites pretread with DFO proliferate normally in trophoblastic cells demonstrating that DFO itself does not interfere with parasite proliferation. Additionally, T. gondii infection induced enhancement in transferrin receptor mRNA expression levels in trophoblastic cells, and the expression was higher in HTR-8/SVneo compared with BeWo. Finally, DFO-treatment was able to reduce the parasite replication in villous explants. Thus, the iron supplementation can be a double-edged sword; in one hand, it could improve the supplement of an essential ion to embryo/fetus development, and on the other hand, could improve the parasite proliferation enhancing the risk of congenital infection.
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Funding
This work was supported by Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG), and Conselho Nacional de Pesquisa Científica e Tecnológica (CNPq). In addition, this study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brazil (CAPES)—Finance Code 001. N.M. Silva, E.A.V. Ferro and B.F. Barbosa are researchers fellows from CNPq.
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Fig S1
Intracellular iron levels in the cell lines. BeWo (3 × 104/200 μL/well) and HTR-8/SVneo (1 × 104/200 μL/well) cells were cultured for 24 h, infected or not with T. gondii (1:1), treated with DFO (250 μM) or incubated with medium (medium) only and submitted to iron measurement by analytical method. The data were shown as iron levels (μg/dL) present in the cell and expressed as mean ± SEM.) (PNG 62 kb)
Fig S2
Viability of infected cells treated with Holo-Transferrin or DFO. (PNG 126 kb)BeWo (3 × 104/200 μL/well), HTR-8/SVneo (1 × 104/200 μL/well) and HeLa (3 × 104/200 μL/well) cells were cultured for 24 h, infected with T. gondii (1:1), treated with human Holo-Transferrin (250 μg/mL), DFO (the median inhibitory concentration (IC50) specific for each cell line) or with complete medium (control), incubated for additional 24 h, and submitted to MTT analyzes. The data were shown as the percentage of viable cells (cell viability) for BeWo (A), HTR-8/SVneo (B) and HeLa (C) cells in relation to untreated cells (medium - 100% of cell viability) and expressed as mean ± SEM.
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Almeida, M.P.O., Ferro, E.A.V., Briceño, M.P.P. et al. Susceptibility of human villous (BeWo) and extravillous (HTR-8/SVneo) trophoblast cells to Toxoplasma gondii infection is modulated by intracellular iron availability. Parasitol Res 118, 1559–1572 (2019). https://doi.org/10.1007/s00436-019-06257-2
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DOI: https://doi.org/10.1007/s00436-019-06257-2