Study On The Expression Of MAGE-3 And NY-ESO-1 In Human HCC And The Corresponding Synthetic Peptide As Anti-tumor Vaccine

Liver cancer is one of the most detrimental cancer and with a relatively high morbidity in Asian country. Now the treatment of liver cancer is mainly dependent on surgical resection. Despite the rapid development of hepatectomy and the improvement on the 5-year-surval rate of small cancer with a diameter less than 3 centimeter in the past 50 years, the outcome of liver cancer is unsatisfactory. The poor prognosis is mainly due to recurrence and metastasis post-operation. For these patients and those who can not take the operation for different reasons, immunotherapy rise the hope to improve the treatment.Since the discovery of tumor specific antigen by Foley and his collegers in 50's of 20 century, the development of basic immunology is more rapidly in past 20 years, especially on the identification of different tumor specific antigen and well understanding of the mechanism of immune response. Many research work and clinical trials on tumor vaccines on the base of epitopes about tumor antigen have been conducted, and cancer/testis (CT) antigens are pay much more attention for their best tumor specific and high immunogenicity.CT antigens are expressed in variety cancers, include melanoma, breast cancer, lung cancer, ovarian cancer but few report about hepatocellular carcinoma. We study the expression of two CT antigen MAGE-3 and NY-ESO-1 in hepatocellular carcinoma, and analyzed the clinical features. Further more we usesynthetic peptide coding by the epitope of these antigens and load on DC to induce special CTL witch lyses the target cells loading with the same antigen.Part I Study on the expression of MAGE-3 and NY-ESO-1 in humanHCC and analysis with clinical featuresObjective: To investigate the expression of cancer/testis antigen MAGE-3 and NY-ESO-1 in human hepatocellular carcinoma and evaluate the positive expression with the clinical features. Method: 60 specimen by hepatectomy with liver cancer were detected using RT-PCR. Results: MAGE-3 mRNA were detected in 64.81% of 54 HCC, and NY-ESO-1 in 32% of 25 HCC. The expression of such genes has no responding with clinical-pathological features. Conclusion:MAGE-3 and NY-ESO-1 express highly in HCC and the expression of these genes can not indicate the prognosis.Part II .Culture and function examination ofdendritic cells in vitroObjective: culture the dendritic cells in vitro as antigen presenting cell of peptide vaccine. Method: Culture the PBMC with 1640 containing GM-CSF and IL-4,identify the phenotype with flow cytometry and exam the bioactivity with IL-12/INF-r secretion. Results: DC can be cultured from monocyte and has the bioactivity. Conclusion: mature DC can be cultured from monocyte and use as vector of peptide vaccine.Part lH.Synthetic peptide coding with the epitope of MAGE-3and NY-ESO-1 to induce special CTL in vitroObjective: To evaluate the synthetic peptide coding with the epitope of MAGE-3 and NY-ESO-1 inducing special CTL in vitro. Method: Mature DC loading with the synthetic peptide coding with the epitope of MAGE-3 and NY-ESO-1 to stimulate autologous T cells in order to induce special CTL and detect the special lysis of corresponding target cells by LDH. Results: synthetic peptide can induce special CTL and lysis the corresponding target cells bearing the same epitope. Conclusion: Synthetic peptide coding with the epitope of MAGE-3 and NY-ESO-1 can induce special CTL and take as the candidate of anti-tumor peptide vaccine.