Aim: To construct the superantigen SEA (D227A) expression vector modulated by cis-acting element of AFP and express it specifically on AFP(+) hepatocellular carcinoma (HCC) cells.
Methods: The enhancer and promoter of AFP gene, SEA (D227A) cDNA and linker-CD80tm were amplified by PCR. The gene segments mentioned above were cloned into the multiple cloning sites of retrovirus vector pLXSN to construct an attenuated superantigen expression vector pLXSN-SEA (D227A)-linker-CD80tm modulated by cis-acting element of AFP gene. AFP(+) or AFP(-) HCCs were transfected by the superantigen expression vector through lipofectamine mediation. SEA expression was detected by RT-PCR and indirect immunofluorescence staining.
Results: Promoter, enhancer, SEA (D227A) and linker-CD80tm had been successfully cloned into the multiple cloning site of retrovirus vector pLXSN RT-PCR and indirect immunofluorescence assay were used to prove that SEA (D227A) could specifically express in the AFP(+) HCCs.
Conclusion: The successful construction and expression of hepatocellular carcinoma-specific superantigen SEA (D227A) gene expression vector lay the foundation for enhancing immunotherapy of hepatocellular carcinoma.