Institute and Cathedra of Cell Biology, Histology and Embryology, Health Science Faculty, Universidad Nacional de Córdoba - INICSA (CONICET), Ciudad Universitaria Córdoba, Córdoba, Argentina.
Instituto Académico Pedagógico de Ciencias Humanas (IAPCH), Universidad Nacional de Villa María (UNVM), Villa María, Córdoba, Argentina.
Am J Reprod Immunol. 2023 Oct;90(4):e13777. doi: 10.1111/aji.13777.
Congenital Trypanosoma cruzi (T. cruzi) infection has been associated with changes in the levels of TNF-α and IFN-γ during the pregnancy. Therefore, we propose to study the participation and dynamics of proinflammatory cytokines in the infection process of placental explants infected by T. cruzi in vitro.
Chorionic villous explants (CVE) obtained of human term placentas (n = 8) from normal pregnancies were cultured with 10 trypomastigotes/mL of Tulahuen strain DTU VI for 0, 2, 4, 16, 24, 48 and 72 h. Explants were treated with sulfasalazine (SULF) (5 mM) and N-acetyl-cysteine (NAC) (15 mM), as inhibitors molecules of NF-κB pathway, or LPS (1 μg/mL) for 24 and 72 h p.i. Motile trypomastigotes were counted in culture supernatants. Immunohistochemistry and ELISA for TNF-α, IFN-γ, IL-1β, IL-4, and IL-10 were performed in CVE and culture supernatants respectively. The parasite load was measured by RT-qPCR.
T. cruzi invades the chorionic villi from 4 h p.i. increasing significantly its DNA at 48 and 72 h p.i. of culture (parasite multiplication phase). They were detected in stromal cells, which was related to elevation of TNF-α, IL-1β, IFN-γ, and IL-10. The inhibition of NF-κB activity in the explants decreased the production of the analyzed cytokines, showing elevated levels of T. cruzi DNA during the multiplication phase of the parasite.
Placental tissue modifies the secretion of pro-inflammatory cytokines during the phase of parasite multiplication, but not during the invasion phase, which in turns modifies the level of infection via the signaling pathway NF-κB.
先天性克氏锥虫（T. cruzi）感染与怀孕期间 TNF-α 和 IFN-γ 水平的变化有关。因此，我们建议研究促炎细胞因子在体外感染 T. cruzi 的胎盘组织中的参与和动态变化。
从正常妊娠的足月胎盘（n=8）中获得绒毛膜绒毛组织（CVE），用 10 个 Tulahuen 株 DTU VI 的锥虫感染，浓度为 10 个/ml，感染 0、2、4、16、24、48 和 72 小时。用 NF-κB 通路抑制剂柳氮磺胺吡啶（SULF）（5mM）和 N-乙酰半胱氨酸（NAC）（15mM）以及脂多糖（LPS）（1μg/ml）处理 24 和 72 小时。在培养上清液中计数运动性锥虫。对 CVE 和培养上清液进行 TNF-α、IFN-γ、IL-1β、IL-4 和 IL-10 的免疫组织化学和 ELISA 检测。通过 RT-qPCR 测量寄生虫负荷。
T. cruzi 从感染后 4 小时开始侵入绒毛，在 48 和 72 小时的培养物中（寄生虫增殖期）其 DNA 显著增加。在基质细胞中检测到它们，这与 TNF-α、IL-1β、IFN-γ 和 IL-10 的升高有关。在组织中抑制 NF-κB 活性降低了分析细胞因子的产生，显示寄生虫增殖期寄生虫 DNA 水平升高。
胎盘组织在寄生虫增殖期改变促炎细胞因子的分泌，但不在入侵期改变，这反过来通过 NF-κB 信号通路改变感染水平。
