摘要
将热稳定抗原(Heat-stable Antigen,简称HSA)的cDNA与质粒载体(PcDNA3)连接,构建成真核表达载体,通过电穿孔的方法将重组质粒导入到小鼠淋巴瘤细胞EL-4中,使其表达抗性基因和HSA分子,通过高剂量G418(400μg/ml)选择培养和有限稀释法克隆,获得高比例表达HSA的阳性细胞克隆,以提供活化T细胞所必需的协同刺激信号,从而诱导宿主体内有效的抗肿瘤免疫应答。实验发现:表达HSA的EL-4淋巴瘤细胞在同系小鼠(C57BL/6)体内的致瘤性明显较野生型肿瘤弱。HSA^+EL-4瘤细胞经丝裂霉素灭活后,腹腔免疫同系小鼠后可获得对低剂量(2×10~3/鼠)野生型瘤细胞EL-4攻击的免疫保护效应。用HSA^+瘤细胞灭活后作为瘤苗治疗早期带瘤动物显示一定的治疗效果。
A recombinant eukaryotic expression vector containing HSA(Heat-stable Antigen) cDNA and PcDNA 3 plasmid was constructed and then transfected into mouse lymphoma cell line---EL-4 by electroporation. The transfected tumor cells were selected in RPM11640 containing G418 (400μg/ml).HSA expression was detected by FACS using indirect immunofluorescene technique with HSA mAb (Ml/69).To obtain high expression of HSA'EL-4 cells ,the transfected cells were recloned by limiting dilution. In animal experiments, we found that the tumorigenicity of HSA+ EL-4 is weaker than HSA- EL-4(EL-4 or EL-4-v). The size of HSA+ EL-4 tumors were significantly smaller than that of HSA- EL-4 tumors. The tumor growth speed and survival time of tumor-bearing mice are also different. A protective effect against the subsequently challenge with low dose (2 × 1 03/mouse) wild type tumor cells was found in immunized mice with inactivated HSA+ EL-4 tumor cells but not in those animals immunized with HSA- EL-4 tumor cells. Moreover, HSA+ EL-4 could be used as tumor vaccine to cure the established tumor at the initial stage.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
1996年第4期272-276,共5页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(39570793)
