Anti-tumor Immune Response Of Human Dendritic Cell Vaccine Encoding With EBV-LMP2

Objective 1. To generate human peripheral blood mononuclear cells (PBMC) derived dendritic cell (DC) induced by cytokine combinations of rhGM-CSF, IL-4 and TNF-α, and develop a DC-based vaccine (rAd-LMP2-DC) by adenovirally transfection with EBV-LPM2 gene. Study its biological charactetistics and detect its anti-tumor immune functions of cytotoxity T lymphocytes (CTL) activated by rAd-LMP2-DC vaccine in vitro, and observe the the immunotherapeutic and immunopreventive effects of rAd-LMP2-DC vaccines on human NPC cells (CNE-2) in SCID mice. 2. To explore an efficient method to induce mature DC by calcium ionophore in serum-free medium and compare its anti-tumor activity with that of DC induced by conventional cytokine combinations.Method 1. Preparation of rAd-LMP2-DC vaccine: The monocytes were induced by GM-CSF and IL-4 for 7 days as immature DCs and then transfected with LMP2 gene by the vector of recombinant adenoviruses. The cells were cultured with GM-CSF and TNF-a for additional 2 days and then harvested as mature rAd-LMP2-DC vaccine. The morphology of DC was observed dynamically. The morphological features of rAd-LMP2-DCs were observed under inverted phase contrast microscope and scanning electron microscope. The expression of DC typical phenotypes of CD80, CD86, CD83 and positive expression of LMP2 in the...