Shimazaki J, Tsubota K, Tsuchida F, Sato T, Hagihara M, Tsuji K
Department of Ophthalmology, Tokyo Dental College, Chiba-ken, Japan.
Nippon Ganka Gakkai Zasshi. 1995 Nov;99(11):1248-53.
DNA-HLA (human leukocyte antigen) typing was performed for ocular tissues in order to determine the usefulness of the method in corneal transplantation. Each type of ocular tissue was dissected from eye bank eyes (n = 3). DNA was extracted, and HLA-DRB1, DQB1, and DPB1 genes were amplified using PCR (polymerase chain reaction) method. DNA can be extracted and amplified from each ocular tissue except for the crystalline lens, but DNA from iris and choroid can be amplified only after diluting the samples 10 to 100 times. HLA class II antigens were successfully determined in these ocular tissues by the RFLP (restriction fragment length polymorphism) method. The method was applied to the corneal tissues of donor and recipient used for the corneal transplantation (n = 7). HLA class II antigens can be determined in both the donor and recipient corneal samples, and the results of recipients' HLA typing were the same as those determined using the blood samples. These results indicate that the DNA-HLA typing can be used for ocular tissues. Since DNA-HLA typing is more accurate than the conventional serological typing, the method is promising for the study of HLA class II typing in corneal transplantation.
为了确定该方法在角膜移植中的实用性，对眼组织进行了DNA - HLA（人类白细胞抗原）分型。从眼库的眼睛（n = 3）中解剖出每种类型的眼组织。提取DNA，并使用PCR（聚合酶链反应）方法扩增HLA - DRB1、DQB1和DPB1基因。除晶状体外，每种眼组织都可以提取和扩增DNA，但虹膜和脉络膜的DNA只有在将样品稀释10至100倍后才能扩增。通过RFLP（限制性片段长度多态性）方法成功测定了这些眼组织中的HLA II类抗原。该方法应用于用于角膜移植的供体和受体的角膜组织（n = 7）。供体和受体角膜样本中均可测定HLA II类抗原，受体的HLA分型结果与使用血液样本测定的结果相同。这些结果表明DNA - HLA分型可用于眼组织。由于DNA - HLA分型比传统的血清学分型更准确，该方法在角膜移植中HLA II类分型研究方面很有前景。
