成人胰岛细胞移植治疗1型糖尿病

Clinical islet transplantation treating type 1 diabetes mellitus

目的探讨新型成人胰岛细胞分离、纯化技术分离的成人胰岛细胞移植治疗1型糖尿病的安全性与有效性。方法采用全氟化碳液（PFC）和UW液双层冷藏胰腺，Liberase酶消化，COBE2991型专用胰岛细胞分离机分离及连续密度梯度纯化，获取高纯度与高活性的胰岛细胞。通过上腹部小切口，胃右静脉或脐静脉插管，将经短期培养的胰岛细胞经门静脉移植到11例1型糖尿病患者肝脏内。采用达利珠单抗或阿来佐单抗进行诱导，西罗莫司和他克莫司联用的方案预防排斥反应，术后观察胰岛素使用情况，监测血糖、C-肽与糖化血红蛋白水平以及肝功能、肾功能。结果45个胰腺中，42个成功分离出胰岛细胞，其数量平均为28．5×10^4胰岛当量（IEQ）／个，纯度为95．7％，活率为93．2％，胰岛素释放试验刺激指数为2．43。11例1型糖尿病患者共行胰岛细胞移植20次，其中移植1次4例，2次5例，3次2例，每次移植胰岛细胞数平均为11200 IEQ／kg。术后随访6个月至4年，6例完全撤除胰岛素，2例胰岛素用量较术前减少80％，3例减少50％。术后血糖稳定维持在正常水平，C-肽均超过0．166nmol／L，糖化血红蛋白基本正常，肝、肾功能正常，未发生与胰岛细胞输注相关的并发症。结论新型成人胰岛细胞分离、纯化方法可靠，采用该技术分离、纯化的成人胰岛细胞移植治疗1型糖尿病临床效果较好。

Objective To establish a new technique isolating pancreatic islet of Langerhans and evaluate the clinical efficacy and safety of adult islet transplantation in the treatment of type 1 diabetes mellitus （T1DM）. Methods Pancreases were stored using the “2-layer method” of the oxygenated perfluorochemical （PFC） and UW solution. The pancreases were digested with Liberase collagenase enzyme and purified using continuous gradients of Ficoll-diatrizoic acid on a refrigerated COBE 2991 centrifuge to separate the islets. Cultured islets were infused by surgical approach to the liver via portal vasculature. Clinical metabolic data such as blood glucose, dose of insulin, C-peptide, HbAlc, liver function and renal function, were determined and compared with the pre-transplant data. Results Islets of Langerhans were isolated successfully in 42 of 45 pancreases. The average of islet yield was 285 000 islet equivalents （IEQ）. Islet purity and viability were 95.7% and 93.2% respectively. The stimulation index （SI） as assessing function of human islet was 2.43. Twenty clinical islet transplant infusions were carried out in 11 subjects with T1DM. Five recipients received 2 islet infusions, 2 patients had 3 transplants, and 4 patients received 1 transplant only. The average islet mass for infusion was 11200 IEQ/kg. The immunosuppressive strategies for islet transplantation： campath-lH induction, sirolimus and low-dose tacrolimus maintenance therapy without glucocorticoid. During the follow-up of 6 months to 4 years, 6 recipients had insulin independence, and the dosage of insulin was decreased by 80 % in 2 patients and by 50 % in 3 patients respectively after islet transplantation. The levels of blood glucose and HbAlc, liver and renal function were normalized throughout the follow-up period. All patients were positive for C-peptide after islet transplantation. No adverse effects and complications related to islet infusion procedure were found. Conclusion New technique has been proved to be suitable for isolating pancreatic islet of Langerhans. Adult islet transplantation can be used as an effective and safe way for treating T1DM.