Acquirement Of Human Heat Shock Protein 70-tumor Peptide Complex(HSP70-PCs) Vaccine By Genetic Engineering And Its Evaluation In Vitro

Aim To express human heat shock protein 70(HSP70)in E.coli and acquire HSP70-tumor peptide complex(HSP70-PCs) vaccine,then evaluate its efficacy in vitro to explore new strategies of treating cancer. Methords The cDNA fragment encoding HSP70 was amplified by PCR ,the PCR products were cloned into vector pET30a.The recombinant plasmid, pET30a -HSP70 was transformed to E.coli strain BL21,then the product of HSP70 was purified. LS 174 T human colon cancer cells were cultured and were disrupted to acquire the tumor mixed peptide, HSP70 and the tumor mixed peptide were conncted in vitro to aquire HSP-PCs.Lymphocytes were stimulated by DCs plused with HSP70-PCs,DCs plused with HSP70,DCs plused with tumor mixed peptide and HSP70-PCs directly,the exosomatic specific cytotoxin against LS 174 T were measured by MTT. Results 1. HSP70 was successfully acquired though genetic engineering ,and the purity of HSP70 after purification was more than 90%. 2. The tumor mixed peptide and HSP-PCs Were acquired.3. The CTL immunized with HSP70-PCs-DC were remarkably higher than the others (p<0.01),while the others have no significant difference(p>0.05). Conclusion 1.Vaccine HSP70-PCs can induce specific CTL cellular immune response,while establish the experimental base for its treatment for cancer. 2. The effective and broad of application of HSP70-PCs which was limited by HSP70 preparation could be solved by this study,and it could be offer a new strategy to cancer immunotherapy.