Experimental Study on Purification of Heat Shock Protein 70 Antigen Peptide Complex from Human Lung Adenocarcinoma GLC-82 Cell and the Anti-tumor Effect of Cytotoxic T Lymphoc-yte acted by Heat Shock Protein

Objective 　To evaluate the purification methods of heat shock protein 70 antigen peptide complex ( HAC-70) f rom human lung adenocarcinoma GLC-82 cell, and to explore the phenotype varieties and anti-tumor function of CTL as well as its suspension induced by HAC-70. Methods 　GLC-82 cells were cultured with 43 ℃for 30 minutes to induce over-expression of HSP. The induced HAC-70 was purified with ion exchange chromatography. The purified HAC-70 was analyzed quantity and quality with SDS-PA GE and EL ISA methods. PBMC were activated by HAC-70. CTL phenotypes were determined by lymphocyte subgroup test kit . The anti-tumor effects of CTL and it s suspension on GLC-82 were determined with MTT method. Results 　Ion exchange chromatography was successfully applied to purify HAC-70 from GLC-82. HAC-70 f rom heat-treated GLC-82 were over expressed with a rate of 75μg/ ml GLC-82. The positive rates of CD3 + 、CD4 + 、and CD4 + / CD8 + T cells induced by heat-treated group were much higher than cont rols. The best tumor-killer effect s of CTL were obtained in HAC-70 group while the target2effect ratio was 50∶1. Apoptosis of GLC-82 were induced by suspension of CTL in all groups. Conclusion 　Purification of HAC-70 from lung adenocarcinoma GLC-82 cells by ion exchange chromatography is simple and feasible. CD3 + and CD8 + T cells are increased and CD4 + / CD8 + ratio is reversed by activated HAC-70. The tumor-killer function of HAC-70-induced CTL and it s suspension is confirmed and the result s will be of valuable to the preparation of tumor vaccine as well as it s clinical application.