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In vitro anti tumor activity of altered tumorassociated antigen CEA-610Dpeptide
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Abstract:
Objective:To investigated the in vivo efficacy of altered CEA610D peptide vaccine against tumor, so as to provide a basis for its clinical use.Methods:Altered CEA peptide CEA610D, natural CEA peptide CEA605613 and MAGE3 peptide were synthesized by polypeptide solidphase synthesis system. The cytotoxicity T lymphocytes (CTLs) were induced by autologous mixed lymphocytes reaction implused with above peptides; proliferation and cytotoxicity of different CTLs were measured by MTT; phenotypes of the CTLs were detected by flow cytometry; expression of perforin in different CTLs were assayed by RTPCR; IFNγ levels in the supernatants of different CTLs were detected by ELISA.Results: CEA610D peptide more efficiently induced CTL than CEA605613 and MAGE3 peptide did (P<0.05). The number of CD8+T cells in CEA610D group was significantly larger than those in CEA605613 and MAGE3 groups (P<005). When the E∶T was 40∶1, the cytotoxicity of CTLs induced by CEA610D and CEA605613 against CEA+HLAA2+ T84 cells were (56.7±373)% and (51.2±1.86)%, respectively. But the CTL cytotoxicities induced by the three peptides against CEA+HLAA2Lovo cells were all at the background level. Perforin expression and IFNγ level of CTLs in CEA610D group were significantly higher than those in the other two groups (P<0.01).Conclusion:Compared with natural CEA605613 peptide, altered CEA610D peptide can effectively break the immune tolerance to self peptide, and thus has a stronger antitumor activity in vitro.
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Supported by the Research Projects of Shandong Academy of Medical Science (No.200625)
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