骨肉瘤特异性细胞毒T淋巴细胞的诱导及其抗肿瘤特性的研究

Induction of tumor specific cytotoxic T lymphocyte in osteosarcoma and study on its anti tumor effects

目的 诱导骨肉瘤特异性细胞毒T 淋巴细胞(osteosarco ma specific cytotoxic T ly mphocyte ,OSS- CTL) ,观察其体内外的抗肿瘤特性。方法 通过生化方法从HOS- 8603 细胞系中提取纯化骨肉瘤相关抗原(osteosarcom a associated antigen , OSAA) ,并与低剂量IL- 2 、CD 3 单抗协同刺激骨肉瘤致敏的淋巴细胞诱导产生OSS- CTL。结果 OSS- CTL 表型特征为CD 3 + 87-6 % ±6-3 % , CD 4 + 21-7 % ±4-1 % ,CD 8 + 94-7 % ±5-3 % ,CD 11b +1-9 % ±0-7 % ,HLA+ 79-3 % ±8-7 % ,即以CD 8 + T 细胞为主的异质细胞群, 对OSAA 相关的自体骨肉瘤细胞和HOS- 8603 细胞显示高亲和杀伤活性。体内实验表明, OSS-CTL 明显抑制裸鼠皮下骨肉瘤的生长。结论 OSS- CTL

Objective To induce osteosarcoma specific cytotoxic T Lymphocyte (OSS-CTL) in order to observe its anti tumor effects and specificity of OSS-CTL in vitro and in vivo Methods Osteosarcoma associated antigen (OSAA) was separated and purified from human osteosarcoma 8603 (HOS-8603) cell line by biochemical means The osteosarcoma specific CTL were harvested from the peripheral blood lymphocytes (PBL) of the patient suffering from osteosarcoma, which were activated by combining stimulation effects of OSAA, low dose IL-2 and anti-CD 3 monoclonal antibody The anti tumor effect and specificity of OSS-CTL were measured with 3H-TdR method by killing HOS-8603 and autogenous osteosarcoma cell, as well as the cell line of K 562 and SHG-44 The TNF, IL-2, IL-8 expressions of OSS-CTL, TIL and PBL were examined with ELLSA assessment in order to explore the cytotoxic mechanism of OSS-CTL Results The osteosarcoma specific CTL were mainly composed of CD8+CTL heterogenous cell groups, which included CD 3+87 6%±6 3%, CD 4+21 7%±4 1%, CD 8+94 7%±5 3%and CD 11b+1 9%±0 7% The osteosarcoma specific CTL showed highly specific killing activity to HOS-8603 and autogenous osteosarcoma cells In vivo, the suppressing effects of the osteosarcoma specific CTL on nude mice subcutaneous osteosarcoma were more obvious than TIL Conclusion The osteosarcoma specific CTL could be served as a new type of immune cell with high cytotoxic effect for immunotherapy