Sun Cun, Pan Zhi-qiang, Jie Ying, Wang Li, Yan Chao, Wang Ning-li
Beijing Tongren Eye Bank, Beijing Tongren Eye Centre, Capital University of Medical Sciences, Beijing 100730, China.
Zhonghua Yan Ke Za Zhi. 2007 Jul;43(7):594-601.
To investigate pig-rhesus corneal xenografting rejection characterization and evaluate the effect of glucocorticoids on the grafts survival.
Wuzhishan pigs (WZSP) donor corneas were grafted orthotopically into rhesus recipients. 12 rhesus monkeys were divided into two groups: group control and group steroid treatment. Recipients in group steroid treatment were injected with compounded betamethasone 0.5 ml (7 mg/ml) subconjunctivally at the end of the operation, 0.3 ml after operation at 10 days interval for 10 total times. The group control received saline buffer in the same way. All xenografts were evaluated by slit-lamp microscopy. One month after surgery, two recipients in each group were sacrificed for corneal histopathological staining. The concentration of cytokines (IFN, TNF, IL-4, IL-5, IL-10) in the aqueous humor and blood of the recipients, the level of immunoglobulin IgG, IgA, IgM and complement C3, C4 in the blood were detected before surgery and at different timepoint after keratoplasty. The percentage of CD4+, CD8+, CD16+ T lymphocyte was examined by cytometry.
The mean survival time (MST) of xenografts was (15.5 +/- 2.3) days in group control and (182.8 +/- 66.1) days in group steroid treatment. In group control, corneal xenografts showed exudative membrane in the anterior chamber, diffuse edema, and new-forming vessels growing at the peripheral corneal bed. In group steroid, the grafts were transparent and showed slight edema. Histopathological examination showed that in group control, corneal xenografts were infiltrated with inflammatory cells. The corneal endothelia were destroyed and exudative membranes were attached onto the posterior of the grafts. In group steroid treatment, the corneal xenografts showed only minimal edema and inflammatory cells infiltrated. The xenografts endothelium were intact and no exudative membrane appeared. In group control, the concentration of cytokines was increased 3 weeks after surgery than the level before surgery, especially IFN. In group steroid treatment, the changes of the cytokines were mainly the increase of IL-10 and IL-4, the concentration of IFN and TNF was decreased. The immunological analysis showed that the concentration of immunoglobulin IgG, IgA, IgM and complement C3, C4 in group control was mainly increased at 2 - 3 weeks after surgery, in group steroid, the change of the concentration of IgG and C4 was different compared with group control.
Endothelium rejection occurred in pig-rhesus corneal xenografts. The humor immune factors might involve grafts rejection reaction. The glucocorticoid was effective on delaying corneal xenotransplantation rejection.
研究猪 - 恒河猴角膜异种移植排斥反应特征，并评估糖皮质激素对移植物存活的影响。
将五指山猪（WZSP）供体角膜原位移植到恒河猴受体。12只恒河猴分为两组：对照组和类固醇治疗组。类固醇治疗组的受体在手术结束时结膜下注射0.5 ml复方倍他米松（7 mg/ml），术后10天间隔注射0.3 ml，共注射10次。对照组以同样方式接受生理盐水缓冲液。所有异种移植物均通过裂隙灯显微镜评估。术后1个月，每组处死2只受体进行角膜组织病理学染色。在手术前及角膜移植术后不同时间点，检测受体房水和血液中细胞因子（IFN、TNF、IL - 4、IL - 5、IL - 10）的浓度、血液中免疫球蛋白IgG、IgA、IgM和补体C3、C4的水平。通过细胞计数法检测CD4 +、CD8 +、CD16 + T淋巴细胞的百分比。
对照组异种移植物的平均存活时间（MST）为（15.5±2.3）天，类固醇治疗组为（182.8±66.1）天。对照组中，角膜异种移植物在前房显示渗出膜、弥漫性水肿，周边角膜床有新生血管生长。类固醇治疗组中，移植物透明，有轻微水肿。组织病理学检查显示，对照组角膜异种移植物有炎性细胞浸润。角膜内皮被破坏，渗出膜附着在移植物后部。类固醇治疗组中，角膜异种移植物仅显示轻微水肿，有少量炎性细胞浸润。异种移植物内皮完整，无渗出膜出现。对照组中，术后3周细胞因子浓度比手术前升高，尤其是IFN。类固醇治疗组中，细胞因子的变化主要是IL - 10和IL - 4增加，IFN和TNF浓度降低。免疫学分析显示，对照组中免疫球蛋白IgG、IgA、IgM和补体C3、C4的浓度主要在术后2 - 3周升高，类固醇治疗组中，IgG和C4浓度的变化与对照组不同。
猪 - 恒河猴角膜异种移植发生内皮排斥反应。体液免疫因子可能参与移植物排斥反应。糖皮质激素对延迟角膜异种移植排斥有效。
