Induction of cytotoxic T lymphocytes from the hepatocellular carcinoma patients by using MAGE-3- gene-derived and HLA-A2-restricted peptide / 中国普通外科杂志

ABSTRACT

Objective To explore the possibility of immune therapy for hepatocellular carcinoma (HCC) with MAGE-3 antigen peptide (FLWGPRALV). Methods The expression of HLA-A2 in HCC patients were examined with microcytotoxicity assay .The mRNA of MAGE-3 genes in the tumor tissues were detected by RT-PCR, the CD8 positive T lympocytes were separated as effector cells with immunobeads from peripheral blood mononuclear cells (PBMCs) of those patients. The irradiated autologous CD8 negative PBMC were pulsed with MAGE-3 peptide as APC. Some effector cells were co-cultured with APC, and some not as the control. After 14 days′ culture with IL-2, the frequency of effector cells secreting IFN-? in response to MAGE-3 peptide were monitored by IFN-? secretion assay. The hepatocellular carcinoma cell line HLE was used as the target cells.ResultsNine HCC patients were HLA-A2 positive in 25 HCC patients, and 3 of them were MAGE-3 mRNA positive. After two weeks′ culture, the effector cells increased by 4-6 folds. The frequency of CD8 positive cells secreting IFN-? in response to MAGE-3 peptide was 22.0% in 3 MAGE-3 mRNA positive patients(n=3),and 0.5% in 6 negative patients( n=6) (?