Song Hyun Beom, Jun Hyoung-Oh, Kim Jin Hyoung, Lee Young-Ha, Choi Min-Ho, Kim Jeong Hun
Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Biomedical Research Institute, Seoul National University Hospital, Seoul, Republic of Korea.
Department of Biomedical Sciences, College of Medicine, Seoul National University, Seoul, Republic of Korea.
PLoS One. 2017 Apr 13;12(4):e0175159. doi: 10.1371/journal.pone.0175159. eCollection 2017.
Ocular toxoplasmosis is mediated by monocytes infected with Toxoplasma gondii that are disseminated to target organs. Although infected monocytes can easily access to outer blood-retinal barrier due to leaky choroidal vasculatures, not much is known about the effect of T. gondii-infected monocytes on outer blood-retinal barrier. We prepared human monocytes, THP-1, infected with T. gondii and human retinal pigment epithelial cells, ARPE-19, grown on transwells as an in vitro model of outer blood-retinal barrier. Exposure to infected monocytes resulted in disruption of tight junction protein, ZO-1, and decrease in transepithelial electrical resistance of retinal pigment epithelium. Supernatants alone separated from infected monocytes also decreased transepithelial electrical resistance and disrupted tight junction protein. Further investigation revealed that the supernatants could activate focal adhesion kinase (FAK) signaling in retinal pigment epithelium and the disruption was attenuated by FAK inhibitor. The disrupted barrier was partly restored by blocking CXCL8, a FAK activating factor secreted by infected monocytes. In this study, we demonstrated that monocytes infected with T. gondii can disrupt outer blood-retinal barrier, which is mediated by paracrinely activated FAK signaling. FAK signaling can be a target of therapeutic approach to prevent negative influence of infected monocytes on outer blood-retinal barrier.
眼弓形虫病是由感染刚地弓形虫的单核细胞介导的,这些单核细胞会扩散至靶器官。尽管由于脉络膜血管渗漏,受感染的单核细胞能够轻易进入外血视网膜屏障,但关于刚地弓形虫感染的单核细胞对外血视网膜屏障的影响,我们所知甚少。我们制备了感染刚地弓形虫的人单核细胞THP-1,以及在Transwell上生长的人视网膜色素上皮细胞ARPE-19,以此作为外血视网膜屏障的体外模型。暴露于受感染的单核细胞会导致紧密连接蛋白ZO-1的破坏,以及视网膜色素上皮跨上皮电阻的降低。仅从受感染的单核细胞中分离出的上清液也会降低跨上皮电阻并破坏紧密连接蛋白。进一步研究表明,上清液可激活视网膜色素上皮中的粘着斑激酶(FAK)信号传导,而FAK抑制剂可减弱这种破坏。通过阻断CXCL8(一种由受感染的单核细胞分泌的FAK激活因子),可部分恢复被破坏的屏障。在本研究中,我们证明感染刚地弓形虫的单核细胞可破坏外血视网膜屏障,这是由旁分泌激活的FAK信号传导介导的。FAK信号传导可能是一种治疗方法的靶点,以防止受感染的单核细胞对外血视网膜屏障产生负面影响。