Initial study of α1,3-galactosyltransferase gene-knockout/CD46 pig full-thickness corneal xenografts in rhesus monkeys
Initial study of α1,3-galactosyltransferase gene-knockout/CD46 pig full-thickness corneal xenografts in rhesus monkeys
复制标题
α1,3-半乳糖基转移酶基因敲除/CD46猪全层角膜异种移植物在恒河猴中的初步研究
DOI：
10.1111/xen.12282
复制
发表时间：
2017-01-01
影响因子：
3.9
通讯作者：
Pan, Zhiqiang
中科院分区：
文献类型：
--
作者：
Dong, Xiaojuan;Hara, Hidetaka;Pan, Zhiqiang
背景：为了研究表达人类补体调节蛋白（GTKO/CD46猪）的α1,3-半乳糖基转移酶基因敲除（GTKO）猪在恒河猴中进行全层角膜异种移植后的移植物存活率。方法：将来自野生型（WT；n=4）和GTKO/CD46的全层角膜移植到恒河猴（n=10） (n= 4) 猪或猴子 (n= 2)。所有受体猴在移植后均接受倍他米松结膜下注射。通过裂隙灯评估角膜移植物。移植后 6 个月测试移植物的组织病理学、免疫组织化学和眼房水中的细胞因子浓度。分别采用流式细胞术和ELISA测定抗猪IgM/IgG和抗半乳糖-α1,3-半乳糖(Gal)抗体。结果：WT和GTKO/CD46异种移植物的最长移植物存活分别为157天和171天。 WT 和 GTKO 猪角膜的移植物存活率没有显着差异。两名 WT 受者和所有 GTKO/CD46 移植物受者均出现前粘连。所有异种移植受者均出现角膜后膜，炎症细胞（CD3+ T 淋巴细胞）浸润角膜基质，并且移植后 6 个月房水中的 IL-6 增加。记录了针对 Gal 和/或非 Gal 猪抗原诱导的抗体反应。相比之下，同种异体移植物存活> 180天，没有任何排斥反应，房水中的细胞因子没有增加，也没有检测到引发的血清抗猪抗体。结论：与WT猪角膜相比，α1,3-半乳糖基转移酶基因敲除/CD46猪角膜与移植物存活时间延长或抗体反应降低无关。如果要实现延长角膜异种移植物的存活，那么在角膜异种移植后预防前粘连和角膜后膜的发展似乎很重要。
Background: To investigate graft survival after full--thickness corneal xenotransplantation from alpha 1,3-galactosyltransferase gene-knockout (GTKO) pigs expressing a human complement regulatory protein (GTKO/CD46 pigs) in rhesus monkeys.Methods: Rhesus monkeys (n= 10) were transplanted with full--thickness corneas from wild-type (WT; n= 4) and GTKO/CD46 (n= 4) pigs or from monkeys (n= 2). All recipient monkeys received post-transplant subconjunctival injections of betamethasone. Corneal grafts were evaluated by slit-lamp. Histopathology, immunohistochemistry of the grafts, and cytokine concentrations in the aqueous humor were tested 6 months after transplantation. Anti-pig IgM/IgG and anti-galactose-alpha 1,3-galactose (Gal) antibodies were determined by flow cytometry and ELISA, respectively.Results: The longest graft survival of WT and GTKO/CD46 xenografts was 157 and 171 days, respectively. There was no significant difference in graft survival between WT and GTKO pig corneas. Anterior synechiae occurred in two recipients of WT and all recipients of GTKO/CD46 grafts. All xenograft recipients developed a retrocorneal membrane, inflammatory cells (CD3+ T lymphocytes) infiltrated the corneal stroma, and in the aqueous humor, IL-6 was increased 6 months after transplantation. Induced antibody responses were documented against Gal and/or non-Gal pig antigens. In contrast, allografts survived > 180 days without any rejection, with no increase in cytokines in the aqueous humor, and no elicited serum anti-pig antibodies were detected.Conclusions:alpha 1,3-galactosyltransferase gene-knockout/CD46 pig corneas were not associated with prolonged graft survival or a reduced antibody response compared with WT pig corneas. The prevention of the development of anterior synechiae and a retrocorneal membrane after corneal xenotransplantation would appear to be important if prolonged corneal xenograft survival is to be achieved.