T細胞疫苗誘導異種胰島移植免疫耐受的實驗研究

[摘要] 目的 體外制備供體特異性T細胞疫苗(T cell vaccine)，探討T細胞疫苗免疫(T cell vaccination)誘導異種胰島細胞移植免疫耐受的作用。方法 腹腔注射鏈脲佐菌素誘導建立Balb/c小鼠Ⅰ型糖尿病模型，用酶消化法和密度梯度離心法無菌分離純化Wistar大鼠胰島細胞。制備Balb/c小鼠針對Wistar大鼠的T細胞疫苗:實驗隨機分為三組，G1:糖尿病小鼠對照組;G2:胰島細胞移植組，糖尿病小鼠+胰島細胞移植+1640培養液(對照組);G3:胰島細胞移植加T細胞疫苗免疫組，糖尿病小鼠+胰島細胞+T細胞疫苗(實驗組)，于d1，d7，d14，d21實驗組Balb/c小鼠接種TCV(1×107/mL)，對照組用1640替代。接種前和每次接種后第5d分別進行混合淋巴細胞反應。胰島細胞移植:將Wistar大鼠的胰島按1200～1500IEQ/kg經肝門靜脈導入小鼠體內，移植后觀測血糖和體重變化。結果 異種胰島細胞移植可以誘導I型糖尿病小鼠血糖降至正常水平且維持一段時間，對照組小鼠移植后(5.12±1.36)d即發生排斥，實驗組移植物存活時間達(20.25±3.45)d。結論 TCV能夠有效延長異種胰島移植物的存活時間，是一種潛在誘導異種胰島移植免疫耐受的方法。

[關鍵詞] T細胞疫苗;胰島細胞;異種移植;免疫耐受

[中圖分類號] R349.1 [文獻標識碼] A [文章編號] 1673-9701(2010)04-19-03

The Effects of T Cell Vaccination on Xenogeneic Islet Cell TransplantationHU Chunlei1ZHAO Zhenlin2

1.Shanxi Medical University，Taiyuan 030001，China;2.The Second Hospital of Shanxi Medical University，Taiyuan 030001，China

[Abstract] Objective To explore the the effects of T cell vaccination on xenogeneic islet cell transplantation. Methods Balb/c mice (H2-d) were used as recipients and Wistar rats were used as donors. TypeⅠdiabetes mice model were made by injecting streptozotocin intravenously. Type Ⅰ diabetes mice were randomLy divided into three groups，Group A:Diabetic mice with no transplantation and no TCV， Group B: Islet cell transplantation +RPMI1640 culture medium， Group C(experimental group): Islet cell transplantation+TCV.T cell vaccine was made by using attenuated spleen lymphocytes from Balb/c mice which were challenged with Wistar rat spleen lymphocytes in vitro and were stimulated with Con A. Balb/c mice as recipients were immunized with TCV on the time points of d1，d7，d14，d21 and;RPMI1640 culture medium was used in control group. One-way mixed lymphocyte reaction(MLR) was performed with effector of immunized Balb/c mice peripheric lymphocytes and stimulator of Wistar rat peripheric lymphocytes on the second day of T cell vaccination. Islet cells were isolated from Wistar rat pancreas using the protocol of enzyme digestion and density gradient centrifμgation. Islet cells were transplanted into immunized Balb/c mice at the density of 1200～1500IEQ/kg by the way of portal vein injection. Recipient's blood glucose was monitored everyday and body weight was recorded weekly after transplantation. Results Xenogeneic islet cell transplantation can induce type I diabetic mice blood glucose to normal levels and remain for some time，the control group mice after transplantation(5.12±1.36) days， which happened rejection，the experimental group graft survival time of up to(20.25±3.45)days. Conclusion T cell vaccination can significantly prolong the survival time of xenogeneic pancreatic islet grafts;T cell vaccination is a potential approach to to induce xenogeneic islet transplant tolerance.

[Key words] T cell vaccine;Islet cell;Xenogeneic transplantation;Immune tolerance

異種移植為胰島移植治療I型糖尿病提供了充足的供體，但嚴重的排斥反應限制了這一方法的應用，應用常規的免疫抑制劑不能抑制異種免疫排斥反應，因此如何誘導免疫耐受成為一個重要課題。T細胞疫苗(TCV)是特異性抗原活化的反應性T細胞經過化學或物理方法處理滅活后制成的，是目前免疫耐受研究中較新的領域[1]。……