INDUCTION OF HUMAN HEPATOMA-SPECIFIC IMMUNE RESPONSE BY HEPATOMA-DENDRITIC CELL HYBRIDOMAS

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1) Circulating dendritic cell subsets were analyzed by flow cytometry in patients with various liver diseases. Freshly-isolated peropheral blood mononuclear cells (PBMNCs) were stained with a lineage (lin) cocktail (anti-CD3, CD14, CD16, CD19, CD20, CD56) conjugated with fluorescein isothiocyanate (FITC), phycoerythrin (PE)-conjugated CD11c or CD123, PerCP-conjugated anti-HLA-DR, and APC-conjugated CD40 or CD86. The cells were analyzed on a FACS calibur. DCs were defined as lin-HLA-DR+ cells, and the percentages and absolute numbers of both CD11c+DC (DC1) and CD123+DC (DC2) were counted. The numbers and percentages of circulating dendritic cell subsets (both DC1 and DC2) were fewer in patients with liver diseases compared with nornal controls. Moreover, as liver disease progress from chronic hepatitis to liver cirrhosis, DC1 tended to decrease and DC2 increased. Hepatitis with higher activity showed fewer DC1 cells in peripferal blood and more those cells in the liver, suggesting that DC1 are actively recruited to the liver, possibly chemoattracted by the inflammatory response in the liver. 2)All the attempts to establish the human hepatoma-dendritic cell hybridomas have been failed. We are stll trying to establish the hybridomas using other techniques such as an electroric fusion. 3) Reconstruction of human immune system in the mice was attempted by infusion of 1×10 ^8 cells into NOD/SCID mice, and we found that more than 60% of mononuclear cells in peripheral blood of the infused mice were human PBL 2 weeks after infusion, indicated by CD45 positivity. Human T and B cells were mainly found in lymph nodes in mesenteruim, and human IgG, IgM and IgA were detected in the sera of those mice. However, vaccination in those mice has not been successfully performed, yet.