Azuma Hiromitsu, Yamamoto Tatsuo, Chishima Fumihisa
Department of Obstetrics and Gynecology, Nihon University School of Medicine, Tokyo, Japan.
J Obstet Gynaecol Res. 2016 Jul;42(7):769-75. doi: 10.1111/jog.12993. Epub 2016 Apr 20.
The anti-β2-GPI antibody (aβ2-GPIAb) has been detected in recurrent fetal loss with strong pathogenic activity. The effects of aβ2-GPIAb on cytokine production and aβ2-GPIAb binding sites in first-trimester trophoblast cells were evaluated.
First-trimester trophoblast cells were cultured in 24-well tissue culture plates with immunoglobulin G (IgG) obtained from aβ2-GPIAb-positive and aβ2-GPIAb-negative serum. Cytokines in the cultured supernatant were measured using the suspension array system and enzyme-linked immunosorbent assays. To identify potential binding sites for aβ2-GPIAb, such as toll-like receptors (TLR) 2 or TLR4, we used mouse monoclonal anti-TLR2 and/or anti-TLR4 antibodies to inhibit TLR and then measured cytokine production.
The production of cytokines, such as interleukin-6 and interleukin-8, increased more in response to aβ2-GPIAb-positive IgG than to aβ2-GPIAb-negative IgG in trophoblast cells. The secretion of cytokines from trophoblast cells decreased when the TLR were blocked with mouse monoclonal anti-TLR2 and anti-TLR4 antibodies.
We suspect that aβ2-GPIAb might increase cytokine production by binding to TLR2 or TLR4. The increased cytokine production in response to aβ2-GPIAb might play a role in the increased inflammatory response in the placenta.
在复发性流产中检测到抗β2糖蛋白I抗体（aβ2-GPIAb）具有很强的致病活性。评估了aβ2-GPIAb对孕早期滋养层细胞细胞因子产生及aβ2-GPIAb结合位点的影响。
将孕早期滋养层细胞接种于24孔组织培养板中，分别加入从aβ2-GPIAb阳性和阴性血清中获得的免疫球蛋白G（IgG）进行培养。采用悬浮芯片系统和酶联免疫吸附测定法检测培养上清液中的细胞因子。为了确定aβ2-GPIAb的潜在结合位点，如Toll样受体（TLR）2或TLR4，我们使用小鼠单克隆抗TLR2和/或抗TLR4抗体抑制TLR，然后检测细胞因子的产生。
在滋养层细胞中，与aβ2-GPIAb阴性IgG相比，aβ2-GPIAb阳性IgG刺激后白细胞介素-6和白细胞介素-8等细胞因子的产生增加更为明显。用小鼠单克隆抗TLR2和抗TLR4抗体阻断TLR后，滋养层细胞的细胞因子分泌减少。
我们推测aβ2-GPIAb可能通过与TLR2或TLR4结合增加细胞因子的产生。aβ2-GPIAb刺激导致的细胞因子产生增加可能在胎盘炎症反应增强中起作用。
