Monoclonal antibody (3G5)-defined ganglioside: cell surface marker of corneal keratocytes.
Monoclonal antibody (3G5)-defined ganglioside: cell surface marker of corneal keratocytes.
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DOI：
10.1167/iovs.03-0256
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发表时间：
2004-03
影响因子：
4.4
通讯作者：
B. Stramer;Michael G. K. Kwok;P. Farthing-Nayak;Jae‐Chang Jung;M. Fini;R. Nayak
中科院分区：
文献类型：
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作者：
B. Stramer;Michael G. K. Kwok;P. Farthing-Nayak;Jae‐Chang Jung;M. Fini;R. Nayak
目的评价抗神经节苷脂单克隆抗体3G 5作为角膜基质细胞标志物的价值。方法应用免疫荧光显微镜观察3G 5在角膜基质细胞上的表达。对正常人、牛、猪、兔、大鼠和小鼠角膜的冷冻切片和修复兔角膜进行了研究。对组织培养的人、牛、猪、小鼠和兔角膜细胞进行体外研究。结果3G 5对人、牛、猪、大鼠和兔角膜的冰冻切片染色，但对小鼠角膜不染色，染色模式遵循基质角膜细胞的分布，但不染色上皮和内皮。亚融合的人和牛角膜基质细胞培养物为3G 5阴性。在不更换含血清培养基的情况下保持融合2周的人和牛细胞几乎100%变为3G 5阳性。3G 5抗原组成型表达培养的兔和猪角膜细胞在所有条件下检查。小鼠角膜基质细胞培养物不表达3G 5。3G 5抗原不存在于已愈合20天的兔角膜全层伤口修复区域的成肌纤维细胞上。愈合伤口周围的区域以改变的分布表达3G 5抗原，而3G 5抗原以预期的模式分布在远离伤口的区域。当体外用TGF β 1处理诱导兔角膜细胞表达肌成纤维细胞标记物α-平滑肌肌动蛋白时，3G 5染色模式发生改变。结论：3G 5抗原是鉴定角膜基质细胞和记录其对与伤口修复相关的环境刺激的反应的有用标记物。
PURPOSE To evaluate the anti-ganglioside monoclonal antibody 3G5 as a marker of corneal keratocytes. METHODS 3G5 expression on keratocytes was investigated by immunofluorescence microscopy. Studies were performed on frozen sections of normal human, bovine, porcine, rabbit, rat, and mouse corneas and on repairing rabbit cornea. In vitro studies were performed on tissue-cultured human, bovine, porcine, mouse, and rabbit keratocytes. RESULTS 3G5 stained frozen sections of human, bovine, porcine, rat, and rabbit cornea but not mouse cornea and the staining pattern followed the distribution of stromal keratocytes but did not stain epithelium or endothelium. Subconfluent human and bovine keratocyte cultures were 3G5 negative. Almost 100% of the human and bovine cells that were maintained at confluence without replacement of serum-containing culture medium for 2 weeks became 3G5 positive. The 3G5 antigen was constitutively expressed on cultured rabbit and porcine keratocytes under all conditions examined. Mouse keratocyte cultures did not express 3G5. The 3G5 antigen was not present on myofibroblastic cells in the repairing area of a full-thickness wound in rabbit cornea that had been healing for 20 days. The area surrounding the healing wound expressed 3G5 antigen in an altered distribution, whereas 3G5 antigen was distributed in the expected pattern in areas that were distant from the wound. When rabbit keratocytes were induced to express the myofibroblast marker alpha-smooth muscle actin by treatment with TGFbeta1 in vitro, the pattern of 3G5 staining was altered. CONCLUSIONS The 3G5 antigen is a useful marker for the identification of corneal keratocytes and for documenting their response to environmental stimuli associated with wound repair.