HLA-A0201 restricted epitopes were screened for the hTERT protein, and the dominant epitopes were obtained. Then the immunogenicity of the dominant epitopes were detected, finally 1-2 stronger epitopes were acquired which could induced stronger immune response.

applied bioinformatics software BIMAS and SYFPEITHI to predict hTERT protein HLA-A0201 restricted CTL epitopes. Then screened dominant epitopes according to the score of two kinds of software. Next, applied peptide affinity experiment, lactate dehydrogenase(LDH) release assay as well as human interfron gamma(IFN-γ) ELISPOT epitope screening experiments to veriify dominant epitopes. Finally, acquired the epitopes which had ability to elicit stronger immune respons.

Comprehensive analysis of SYFPEITHI software and BIMAS software for HLA-A0201 restricted CTL antigen epitope prediction of hTERT protein showed that 3 peptides were better than others. They were ILAKFLHWL, ELLRSFFYV and ILSTLLCSL. The result of peptide affinty experiment showed that the fluorescence index(FI) of each peptide was ILAKFLHWL 0.67, ELLRSFFYV 0.66 and ILSTLLCSL 0.90; The result of LDH releasing assay showed that the killing rate of ILAKFLHWL was significantly higher than other peptides and negative peptide(P<0.05); The result of human IFN-γ ELISPOT test showed that the number of spots elicited by ILAKFLHWL epitope was significantly higher than other epitopes(P<0.05).

The PBMCs cells induced by epitope ILAKFLHWL had a most significant killing effect. The epitope ILAKFLHWL could be used to make lung cancer poly-peptide vaccine.