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摘 要:目的 探讨GLC -82细胞中HAC 70的提纯方法及其诱导的CTL表型变化和CTL及其上清液抗瘤效应。方法 GLC- 82 细胞热休克诱导 HSP 表达,离子交换层析提纯 HAC 70, SDS -PAGE 和ELISA法进行定量和定性检测,活化PBMC,T淋巴细胞亚群试剂盒测定 HAC- 70 诱导 CTL细胞表型变化情况,MTT法测定CTL及上清液杀瘤活性。结果 热休克处理能使 GLC- 82 细胞 HAC- 70 表达增加,离子交换层析可提纯 GLC-82 细胞中 HAC- 70。经热休克处理的 HAC -70 诱导 T淋巴细胞,其CD3+、CD~(4+)、CD~(8+)细胞与对照组的CTL细胞阳性率明显提高,其诱导的 CTL杀瘤活性显著提高,且其CTL培养上清液肿瘤杀伤活性最高。结论 离子交换层析可提纯 GLC -82 的 HAC -70;HAC- 70 可使CTL细胞CD~(4+)/CD~(8+)倒置,活化的CTL及上清液有较高的杀瘤活性,为肺癌肿瘤疫苗的制备和临床应用提供了实验依据。
Abstract: Objective To investigate the purification method of HAC 70 in GLC -82 cells and the changes of its CTL phenotype and the anti-tumor effect of CTL and its supernatant. Methods The heat shock induced the expression of HSP in GLC-82 cells. The HAC 70 was purified by ion-exchange chromatography, and detected by SDS-PAGE and ELISA. The PBMCs were activated by TAC and activated by T lymphocyte subsets. Changes, MTT assay CTL and supernatant cytotoxic activity. Results Heat shock treatment can increase the expression of HAC-70 in GLC-82 cells, and HAC-70 can be purified by ion exchange chromatography in GLC-82 cells. The positive rate of CTL in CD3 +, CD4 +, CD8 + cells and control group of HAC-70-induced T lymphocytes was significantly increased after heat shock treatment, and the CTL cytotoxicity was significantly increased , And its CTL culture supernatant tumor killing activity highest. Conclusion Ion exchange chromatography can purify HAC-70 from GLC-82. HAC-70 can upregulate CD 4 / CD 8+ in CTL cells. Activity, provide experimental evidence for the preparation and clinical application of lung cancer vaccine.