Cytopathic effect of Acanthamoeba on human corneal fibroblasts
Cytopathic effect of Acanthamoeba on human corneal fibroblasts
复制标题
棘阿米巴对人角膜成纤维细胞的细胞病变作用
作者：
Noriko Takaoka;S. Yamagami;Seiichi Yokoo;M. Matsubara;K. Yagita
目的阿米巴角膜炎与角膜细胞耗竭有关。我们研究了从阿米巴角膜炎感染的角膜中分离的阿米巴如何在体外与人角膜基质细胞相互作用。方法对6例阿米巴角膜炎患者进行阿米巴分离和基因分型。用离体剥脱的角膜基质评估分离的阿米巴是否可以侵入角膜基质。用MTT法定量评价阿米巴对体外培养的角膜成纤维细胞的致细胞病变作用。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法（TUNEL）和Annexin V染色检测与阿米巴共培养的角膜成纤维细胞中的凋亡细胞。结果18SrDNA序列分析显示，6株阿米巴角膜炎患者的阿米巴均为T4基因型。在离体实验中，阿米巴侵入裸露的角膜基质，并在直接粘附后对人角膜成纤维细胞产生细胞病变作用，但不通过化学介质。通过Annexin V染色证实，所有6种阿米巴均检测到细胞病变效应，角膜成纤维细胞主要通过凋亡死亡。结论从阿米巴角膜炎患者中分离的阿米巴对人角膜成纤维细胞有致细胞病变作用，主要是通过直接粘附后诱导细胞凋亡。我们的研究结果可能为阿米巴角膜炎患者角膜细胞耗竭的病理生理学提供一些线索。
Purpose Acanthamoeba keratitis is associated with keratocyte depletion in humans. We investigated how Acanthamoebae isolated from corneas affected by Acanthamoeba keratitis interacted with human corneal stromal cells in vitro. Methods Acanthamoebae were isolated from 6 patients with Acanthamoeba keratitis and genotyping was done. Whether the isolated Acanthamoebae could invade the corneal stroma was assessed with denuded corneal stroma ex vivo. The cytopathic effect of Acanthamoeba on cultured corneal fibroblasts from donor corneas was quantitatively evaluated by the MTT assay after culture under various conditions. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and Annexin V staining were employed to detect apoptotic cells among the corneal fibroblasts co-cultured with Acanthamoebae. Results All 6 Acanthamoebae isolated from the patients with Acanthamoeba keratitis were shown to have the T4 genotype by 18S rDNA sequence analysis. Acanthamoebae invaded the denuded corneal stroma in the ex vivo experiments and had a cytopathic effect on human corneal fibroblasts after direct adhesion, but not via chemical mediators. A cytopathic effect was detected with all 6 Acanthamoebae and corneal fibroblasts mainly died by apoptosis, as evidenced by Annexin V staining. Conclusions Acanthamoebae isolated from patients with Acanthamoeba keratitis had a cytopathic effect on human corneal fibroblasts, mainly via induction of apoptosis after direct adhesion. Our findings may provide some clues to the pathophysiology of corneal keratocyte depletion in patients with Acanthamoeba keratitis.
影响因子：
4.4
作者：
J. W. Hong;J. Liu;J. S. Lee;R. R. Mohan-R.;D. Woods;Y. He;S. Wilson
通讯作者：
J. W. Hong;J. Liu;J. S. Lee;R. R. Mohan-R.;D. Woods;Y. He;S. Wilson
棘阿米巴在体外人角膜上皮细胞和角膜细胞上的生长。
影响因子：
4.4
作者：
Stopak,SS;Roat,MI;Nauheim,RC;Turgeon,PW;Sossi,G;Kowalski,RP;Thoft,RA
通讯作者：
Thoft,RA