Development of Gene Therapy for Type I Diabetes Using Cytokine Gene and Pancreatic Islet B Cell-Specific Lymphocytes

白细胞介素10 （IL-10）可抑制或加重胰岛素依赖型糖尿病（IDDM）。这种抑制可能是基于IL-10强烈抑制巨噬细胞或Th1淋巴细胞的ifn - γ合成这一事实。基于IL-10诱导MHC II类分子的事实，以及另一项观察，即细胞毒性淋巴细胞同时分泌IL-10和ifn - γ，预计会加重。为了阐明IL-10在胰岛免疫破坏过程中的原位作用，我们首先在NOD菌株（NOD-IL-10-Tg）中产生转基因小鼠（Tg）。通过将大鼠胰高血糖素启动子与小鼠IL-10 cDNA融合，该Tg模型中IL-10的产生可以定位于胰岛A细胞。所产生的IL-10有望通过旁分泌机制对胰岛B细胞发挥作用。该系统的优点是胰岛B细胞不受分子修饰。NOD-IL-10-Tg表现出严重的胰岛素炎和胰岛周围的导管增生伴纤维化。他们甚至在10周大之前就患上了糖尿病。第二种方法是使用胰岛B细胞特异性淋巴细胞。通过从NOD小鼠胰腺浸润中分离出的4个淋巴细胞克隆，研究了IL-10传递到胰岛素炎症部位的可能性。在过继转移模型中，糖尿病通过这些淋巴细胞系转移到年轻NOD小鼠。通过过继性转移IL-10 cDNA转导的淋巴细胞系，可显著降低转移性糖尿病的发生率。此外，野生型淋巴细胞与转导淋巴细胞混合过继转染对该模型糖尿病的发生有较强的抑制作用。本研究显示了IL-10的多种功能，但为发展IL-10转导淋巴细胞基因治疗糖尿病提供了理论基础。

Interleukin 10 (IL-10) can either suppress or aggravate insulin-dependent diabetes mellitis(IDDM). The suppression is possible based on the fact that IL-10 strongly inhibits IFN-gamma synthesis from macrophages or Th1 lymphocytes. The aggravation is expected based on a fact that IL-10 induces MHC Class II molecules, and another observation that cytotoxic lymphocytes secretes IL-10 and IFN-gamma concomitantly. To elucidate the role of IL-10 in situ in the pancreatic islets during the immunological destruction process, we first produced transgenic mice (Tg) in the NOD strain(NOD-IL-10-Tg). By fusing rat glucagon promoter with mouse IL-10 cDNA,the production of IL-10 in this Tg model is rendered to be localized in pancreatic islet A cells. The produced IL-10 is expected to exhibit its function by the paracrine mechanism toward pancreatic islet B cells. The advantage of this system is that pancreatic islet B cells are not molecularly modified. NOD-IL-10-Tg exhibited severe insulitis and ductal proliferation with fibrosis around the islets. They became diabetic even before the age of 10 weeks. The second approach was to use the pancreatic islet B cell-specific lymphocytes. By using 4 lymphocyte clones isloted from pancreatic infiltrates in NOD mice, the possibility of delivering IL-10 to the very locus of insulitis was examined. In the adoptive transfer model, diabetes was transferred to young NOD mice by these lyphocyte cell lines. By adoptively transferring lymphocyte cell lines which were transduced with IL-10 cDNA,the incidence of transferred diabetes was significantly reduced. In addition, adoptive transfer of the mixture of wild type lymphocytes with transduced lymphocytes showed the strong suppression of the occurrence of diabetes in this model. This study showed the various functions of IL-10, but it supplied the theoretical basis for the development of gene therapy for diabetes with lymphocytes transduced with IL-10.