Experimental study of gene therapy for malignant glial cells by transfection with IL-12 gene -using SCID mouse model imitating human being-

1.建立模拟人的实验模型将人单核细胞接种于SCID小鼠腹膜后，3周后检测到人单核细胞。同时给予人IL-2可延长人单核细胞寿命约1~2周。提示人IL-2可激活人NK细胞和T细胞。我们不知道为什么一些SCID小鼠在早期给药后死亡。2.恶性星形细胞瘤和胶质母细胞瘤患者接受β-干扰素治疗后IL-12的表达对于恶性星形细胞瘤或胶质母细胞瘤患者每隔一天接受一次300万单位的β-干扰素免疫治疗。治疗期间检测NK细胞活性、LAK细胞活性和IL-12表达。IL-12基因(p35、p40)转入恶性胶质瘤细胞系后，IL-12基因未能成功导入恶性胶质瘤细胞系。P35和p40亚基都是活动型IL-12所必需的。因此，我们用电穿孔的方法对p35和p40进行了双转染。现在我们研究寻找最可靠的诱导IL-12的方法。另一方面，在体外研究中，IL-12诱导人单个核细胞对人脑胶质瘤细胞株有相当的抗肿瘤作用，甚至与IL-2相比也是如此。然而，为了进行IL-12的临床试验，建立体外模型是非常重要的。

1.Making an Experimental model imitating human beingAfter the intraperitoneal administration of SCID mouse with human mononuclear cells, these human mononuclear cells were detected until 3 weeks. Administration of human IL-2 simultaneously prolonged the life span of human mononuclear cells about 1 or 2 weeks. It suggests that human IL-2 activated human NK cells and T cells. We don't know why some SCID mouse died after the administration early. So it is necessary for experimental model to use the other cytokines.2.IL-12 expression in patients with malignant glioma, having beta-IFN therapyFor patients with malignant astrocytoma or glioblastoma, we underwent immunotherapy with beta-IFN (3 million unit) every other day. During the therapy, NK activity, LAK activity and IL-12 expression were measured. However we cannot find NK activity, LAK activity and IL-12 expresion significantly.3.Transfection of IL-12 gene (p35, p40) into malignant glioma cell lineIL-12 gene could not be successfully introdued into malignant glioma cell line. Both subunits of p35 and p40 are nessesary for active type of IL-12. So we did double transfection of P35 and p40 with electroporation. Now we investigate to find the surest way of induction of IL-12. On the other hand, in vitro study IL-12 induced a considerable antitumor effect from human mononuclear cells on human glioma cell line, even compared with IL-2. However, for the purpose of clinical trial of IL-12, it is important to establish the ex vivo model.