College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin, China.
Diabetologia. 2010 Oct;53(10):2198-204. doi: 10.1007/s00125-010-1829-2. Epub 2010 Jul 1.
AIMS/HYPOTHESIS: Human alpha1-antitrypsin (hAAT) gene therapy prevents type 1 diabetes in a NOD mouse model of diabetes. However, repeated i.p. injections of hAAT into NOD mice leads to fatal anaphylaxis. The aim of the study was to determine if an alternative route of administration avoids anaphylaxis and allows evaluation of hAAT's potential for diabetes prevention and reversal. We also sought to determine if the addition of granulocyte colony-stimulating factor (G-CSF), augments hAAT's capacity to prevent or reverse disease in the NOD mice.
To evaluate hAAT pharmacokinetics, serum hAAT levels were monitored in NOD mice receiving a single dose (2 mg) of hAAT by i.p., s.c. or i.d. injection. For studies of type 1 diabetes prevention and reversal, mice received i.d. hAAT (2 mg/mouse/3 days) for 8 or 10 weeks or hAAT and G-CSF (i.p., 6 microg/day) for 6 weeks. Blood glucose determinations, glucose tolerance testing and insulin tolerance tests were performed.
Both i.p. and s.c. injections resulted in fatal anaphylaxis. The i.d. injection avoided anaphylaxis and i.d. injection of hAAT into 11-week-old NOD mice prevented disease (p = 0.005, AAT vs PBS at 40 weeks of age). Treatment of diabetic NOD mice with hAAT or hAAT plus G-CSF provided long-term (at least 100 days) reversal of diabetes in 50% of treated animals. G-CSF did not enhance the reversal rates of hAAT. Glucose tolerance and insulin levels were normalised in mice with hAAT prevention and reversal.
CONCLUSIONS/INTERPRETATION: Intradermal hAAT prevents and reverses disease in a NOD mouse model of type 1 diabetes without inducing anaphylaxis.
目的/假设：人α1-抗胰蛋白酶（hAAT）基因治疗可预防糖尿病 NOD 小鼠模型中的 1 型糖尿病。然而，反复给 NOD 小鼠腹腔内注射 hAAT 会导致致命的过敏反应。本研究旨在确定替代给药途径是否可避免过敏反应，并评估 hAAT 预防和逆转糖尿病的潜力。我们还试图确定粒细胞集落刺激因子（G-CSF）的添加是否增强了 hAAT 在 NOD 小鼠中预防或逆转疾病的能力。
为了评估 hAAT 的药代动力学，通过腹腔内、皮下或皮内注射单次剂量（2mg）hAAT 后，监测接受注射的 NOD 小鼠的血清 hAAT 水平。为了研究 1 型糖尿病的预防和逆转，小鼠接受皮内注射 hAAT（2mg/只/3 天）8 或 10 周，或 hAAT 和 G-CSF（腹腔内，6μg/天）6 周。进行血糖测定、葡萄糖耐量试验和胰岛素耐量试验。
腹腔内和皮下注射均导致致命的过敏反应。皮内注射可避免过敏反应，并且皮内注射 11 周龄 NOD 小鼠的 hAAT 可预防疾病（p=0.005，40 周龄时 AAT 与 PBS 相比）。用 hAAT 或 hAAT 加 G-CSF 治疗糖尿病 NOD 小鼠可使 50%的治疗动物的糖尿病长期（至少 100 天）逆转。G-CSF 并未增强 hAAT 的逆转率。hAAT 预防和逆转的小鼠的葡萄糖耐量和胰岛素水平正常化。
结论/解释：皮内 hAAT 可预防和逆转糖尿病 NOD 小鼠模型中的 1 型糖尿病，而不会引起过敏反应。
