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人IL 12 (hIL 12 )对鼠源的免疫细胞作用较弱 ,进行hIL 12抗瘤研究缺乏有效的动物模型。为此利用荷人瘤的HuPBL SCID小鼠模型 ,将改良的Winnassay细胞免疫功能分析方法应用于评价hIL 12和hB 1协同抗瘤作用。用商业化的非脂质转基因试剂配制单独的hIL 12、hB7 1或hIL 12联合hB7 1的基因转染液。将上述三种基因转染液分别瞬时转基因入人恶性黑色素瘤A375细胞 ,转基因 30min后 ,按组别 (hIL 12组、hB7 1组、hIL 12 +hB7 1组、rhIL 2对照组、HuPBL对照组和肿瘤对照组 )同人外周血淋巴细胞混合接种于SCID小鼠皮下 ,2 0天后处死实验小鼠 ,测得hIL 12和hB7 1的抑瘤率分别为 74 .0 6 %和 6 6 .98% ,联合作用达 93.4 0 % ,rhIL 2和HuPBL对照组无抗瘤作用。同时 ,在此模型上hIL 12联合hB7 1基因对人大肠癌LoVo和人肺癌SPC的抑制率分别为 98.37%和 97.39%。实验建立的各组HuPBL SCID小鼠 2 0天的外周血都含有人IgG(>0 .5mg/L)和极少量人CD3+ 淋巴细胞 (>0 .5个 /10 0个有核细胞 )。hIL 12、hB7 1和hIL 12 +hB7 1组的瘤灶内人淋巴细胞浸润生长 ,肿瘤细胞残存 ,而rhIL 2和HuPBL对照组的瘤灶内人淋巴细胞“似团样”生长 ,肿瘤生长旺盛。表明此HuPBL SCID小鼠模型上的抗瘤机制为局部免疫反应。结果表明以此动物模型可跨
Human IL 12 (hIL 12) has weak immunocompromised mouse cytotoxicity and no effective animal model for anti-tumor study of hIL 12. For this purpose, a modified Winnassay cellular immune function assay was applied to evaluate the synergistic anti-tumor effect of hIL 12 and hB 1 using a human tumor-bearing HuPBL SCID mouse model. Transfection of hIL 12, hB7 1 or hIL 12 alone with hB7 1 gene transfection was formulated with commercial non-lipid transgenic reagents. Transfection of these three gene transfectants into human malignant melanoma A375 cells were transiently transfected into human A375 cells. Transfection for 30 min followed by transfection of hIL 12 group, hB7 1 group, hIL 12 + hB7 1 group, rhIL 2 control group, HuPBL control group And tumor control group) were inoculated subcutaneously into SCID mice subcutaneously. Twenty mice were sacrificed on day 20. The inhibitory rates of hIL 12 and hB7 1 were 74.06% and 66.98% , The combined effect reached 93.4%, rhIL 2 and HuPBL control group no anti-tumor effect. At the same time, the inhibitory rates of hIL 12 combined with hB7 1 gene on human colorectal cancer LoVo and human lung cancer SPC were 98.37% and 97.39% respectively. The HuPBL SCID mice in each experimental group contained human IgG (> 0.5 mg / L) and a very small amount of human CD3 + lymphocytes (> 0.5 per 10 nucleated cells) on the 20th day. Human lymphocytes in hIL-12, hB7 1 and hIL 12 + hB7 1 groups grew in infiltrating tumor cells, while tumor cells remained. However, human lymphocytes in rhIL-2 and HuPBL control group grew like “clusters” . The anti-tumor mechanism of this HuPBL SCID mouse model is a local immune response. The results show that this animal model can span