AIM: To develop atumor vaccine by fusion of H22 hepatocarcinoma cells and DC, and to study its protective and therapeutical effect against H22 cell.

METHODS: H22-DC vaccine was produced by PEG fusion of H22 and DC induced by cytokine released from splenic mononuclear cells, sorted by CD11c magnetic microbead marker. It was injected through the tail vein of the mice and the H₂₂-DC oncogenesis was detected in the liver, spleen and lung. In order to study the therapeutical and protective effect of H₂₂-DC against tumor H₂₂, two groups were divided: immune group and therapeutic group. Immune group was further divided into P, D, HD and H subgroups, immunized by PBS, DC, H₂₂-DC and inactivated H₂₂, respectively, and attacked by H₂₂ cell. The tumor size, tumor weight, mice survival time and tumor latent period were recorded and statistically analyzed; Therapeutical group was divided into three subgroups of P, D and HD, and attacked by H₂₂, then treated with PBS, DC, and H₂₂-DC, respectively. Pathology and flow cytometry were also applied to study the mechanism how the H₂₂-DC vaccine attacked on the H₂₂ cell.

RESULTS: 1. No oncogenesis was found in spleen, lung and liver after H22-DC injection. 2. Hybrid vaccine immunized mice had strongest CTL activity. 3. In the immune group, latent period was longer in HD subgroup than that in P, H and D subgroup; and tumor size and weight were smaller in HD subgroup than that in P, H and D subgroup. 4. In therapeutic group, tumor size was smaller in HD subgroup than that in P, D subgroup.

CONCLUSION: 1. H22-DC tumor vaccine is safe without oncogenesis in vivo. 2. Hybrid vaccine can stimulate potent specific CTL activity against H22. 3. H22-DC vaccine has distinctive prophylatic effect on tumor H22 and can inhibit the tumor growth.

• Animals
• Cancer Vaccines/therapeutic use
• Carcinoma, Hepatocellular/pathology
• Cell Fusion
• Cell Line
• Dendritic Cells
• Immunotherapy/methods
• Liver Neoplasms/pathology
• Mice
• Mice, Inbred BALB C
• Neoplasms/prevention & control
• Neoplasms/therapy

AIM: To prepare a cancer vaccine (H₂₂-DC) expressing high levels of costimulatory molecules based on fusions of hepatocarcinoma cells (H₂₂) with dendritic cells (DC) of mice and to analyze the biological characteristics and induction of specific CTL activity of H₂₂-DC.

METHODS: DCs were isolated from murine spleen by metrizamide density gradient centrifugation, purified based on its characteristics of semi-adhesion to culture plates and FcR^-, and were cultured in the medium containing GM-CSF and IL-4. A large number of DC were harvested. DCs were then fused with H₂₂ cells by PEG and the fusion cells were marked with CD11c MicroBeads. The H₂₂-DC was sorted with Mimi MACS sorter. The techniques of cell culture, immunocytochemistry and light microscopy were also used to test the characteristics of growth and morphology of H₂₂-DC in vitro. As the immunogen, H₂₂-DC was inoculated subcutaneously into the right armpit of BALB/C mice, and their tumorigenicity in vivo was observed. MTT was used to test the CTL activity of murine spleen in vitro.

RESULTS: DC cells isolated and generated were CD11c+ cells with irregular shape, and highly expressed CD80, CD86 and CD54 molecules. H₂₂ cells were CD11c^- cells with spherical shape and bigger volume, and did not express CD80, CD86 and CD54 molecules. H₂₂-DC was CD11c⁺ cells with bigger volume, being spherical, flat or irregular in shape, and highly expressed CD80, CD86 and CD54 molecules, too. H₂₂-DC was able to divide and proliferate in vitro, but its activity of proliferation was significantly decreased as compared with H₂₂ cells and its growth curve was flatter than H₂₂ cells. After subcutaneous inoculation over 60 d, H₂₂-DC showed no tumorigenecity in mice, which was significantly different from control groups (P < 0.01). The spleen CTL activity against H₂₂ cells in mice implanted with fresh H₂₂-DC was significantly higher than control groups (P < 0.01).

CONCLUSION: H₂₂-DC could significantly stimulate the specific CTL activity of murine spleen, which suggests that the fusion cells have already obtained the function of antigen presenting of parental DC and could present H₂₂ specific antigen which has not been identified yet, and H₂₂-DC could induce antitumor immune response; although simply mixed H₂₂ cells with DC could stimulate the specific CTL activity which could inhibit the growth of tumor in some degree, it could not prevent the generation of tumor. It shows that the DC vaccine is likely to become a helpful approach in immunotherapy of hepatocarcinoma.

• cancer vaccine
• dendritic cells
• hepatocarcinoma cells
• cell fusion
• spleen
• mouse

• Animals
• Antigens, CD/analysis
• B7-1 Antigen/analysis
• B7-2 Antigen
• Cancer Vaccines
• Cell Fusion
• Dendritic Cells/chemistry
• Dendritic Cells/cytology
• Integrin alphaXbeta2/analysis
• Intercellular Adhesion Molecule-1/analysis
• Liver Neoplasms, Experimental/pathology
• Liver Neoplasms, Experimental/prevention & control
• Male
• Membrane Glycoproteins/analysis
• Mice
• Mice, Inbred BALB C
• Spleen/cytology