Alvarado J A, Underwood J L, Green W R, Wu S, Murphy C G, Hwang D G, Moore T E, O'Day D
Department of Ophthalmology, University of California, San Francisco.
Arch Ophthalmol. 1994 Dec;112(12):1601-9. doi: 10.1001/archopht.1994.01090240107034.
To test the hypothesis that the iridocorneal endothelial (ICE) syndrome has a viral origin by comparing the incidence of viral DNA in corneal specimens from patients with the ICE syndrome and from controls.
Thirty-one corneas obtained from 25 patients with the ICE syndrome and six with chronic herpetic keratitis (n = 31) were compared with 30 control specimens obtained from 15 healthy donors and from 15 patients with other, nonviral chronic corneal diseases.
Primer pairs and polymerase chain reaction methods were used to identify and amplify either a segment of the DNA polymerase gene in the case of the herpes simplex and zoster viruses or a region of the nuclear antigen gene for the Epstein-Barr virus. The oligonucleotide amplified by polymerase chain reaction was fully characterized with the use of restriction enzyme, hybridization, and sequence analyses to determine that it contained the expected base pair sequence.
Sixteen of 25 ICE syndrome specimens and four of six herpetic keratitis specimens were positive for herpes simplex virus (HSV) DNA. All nine ICE syndrome specimens tested were negative for the presence of DNA from the herpes zoster or the Epstein-Barr viruses. Controls were uniformly negative for HSV DNA whether they were obtained from ostensibly normal corneas (n = 15) or from corneas with intestinal keratitis, aphakic bullous keratopathy, or keratoconus (n = 15). Tissue samples cut from positive ICE syndrome specimens yielded negative results when retested after the endothelial layer was removed. These findings indicate that localization of HSV DNA is within the endothelium, the tissue primarily involved in the pathogenesis of the ICE syndrome.
Polymerase chain reaction evidence shows that HSV DNA is present in a substantial percentage of ICE syndrome corneal specimens and that HSV-DNA is absent in normal corneas and in corneas from patients with three other chronic corneal diseases. These results provide direct evidence to support our hypothesis that the ICE syndrome has a viral origin. We discussed clinical implications, including possible therapeutic interventions.
通过比较虹膜角膜内皮（ICE）综合征患者与对照组角膜标本中病毒DNA的发生率，检验ICE综合征起源于病毒的假说。
将从25例ICE综合征患者和6例慢性疱疹性角膜炎患者（n = 31）获取的31只角膜，与从15名健康供体以及15例患有其他非病毒性慢性角膜疾病的患者获取的30份对照标本进行比较。
使用引物对和聚合酶链反应方法，在单纯疱疹病毒和带状疱疹病毒的情况下鉴定并扩增DNA聚合酶基因片段，在爱泼斯坦-巴尔病毒的情况下鉴定并扩增核抗原基因区域。通过限制性内切酶、杂交和序列分析对聚合酶链反应扩增的寡核苷酸进行全面表征，以确定其包含预期的碱基对序列。
25份ICE综合征标本中有16份、6份疱疹性角膜炎标本中有4份单纯疱疹病毒（HSV）DNA呈阳性。所有检测的9份ICE综合征标本中，带状疱疹病毒或爱泼斯坦-巴尔病毒的DNA均为阴性。对照标本无论是从表面正常的角膜（n = 15）获取，还是从患有带状角膜病变、无晶状体大泡性角膜病变或圆锥角膜的角膜（n = 15）获取，HSV DNA均呈阴性。从阳性ICE综合征标本切取的组织样本在去除内皮层后重新检测时结果为阴性。这些发现表明HSV DNA定位于内皮，而内皮是ICE综合征发病机制中主要受累的组织。
聚合酶链反应证据表明，相当比例的ICE综合征角膜标本中存在HSV DNA，而正常角膜以及其他三种慢性角膜疾病患者的角膜中不存在HSV DNA。这些结果提供了直接证据支持我们的假说，即ICE综合征起源于病毒。我们讨论了临床意义，包括可能采用的治疗干预措施。
