Anti-cervical Cancer Immunological Effect Induced By Dendritic Cell Transfected With HPV16E6 Antigen Gene

Objective To construct pcDNA3.1/Myc-His(-)A-HPV16E6 recombinant plasmid, transfect it into DC derived from human peripheral blood and prepare HPV16E6-DC vaccine, detect its biological characters, observe the anti-cervical cancer immunological effect of cytotoxicity T lymphocytes (CTL) activated by HPV16E6-DC vaccines in vitro, and to study the immunotherapeutic and immunopreventive effects of HPV16E6-DC vaccines on human CaSki cells in SCID mice.Methods 1. HPV16 DNA was abstracted from the CaSki cells infected with HPV16, HPV16E6 DNA was amplified by PCR, and the pcDNA3.1/Myc-His (-)A-HPV16E6 plasmid with HPV16E6 was constructed by gene recombination technique. 2. Peripheral blood mononuclear cells (PBMC) from healthy volunteers were harvested by Ficoll-Hypaque (ρ=1.077g/mL) gradient centrifugation method, adherent PBMC were isolated after two hours' adhesion, and further incubated in RPMI 1640 medium supplemented with 10% fetal calf serum containing 300ng/mL rhGM-CSF and 25ng/mL rhIL-4. Some of DC on day 7 were stored in liquid nitrogen for two weeks...