Preparation And Pre-clinical Study Of Human Fusion Hepatoma Vaccine

Hepatocellular carcinoma (HCC) is one of the most frequent malignancies in Asian country. Its mortality takes the second place among malignant tumors in China. Although the surgical excision is considered the most effective therapy for early-stage HCC patients, only a very small proportion of patients with an operable primary lesion may transiently benefit from surgical treatment because of a high recurrence rate of cancer after operation. To study and search for an approach of anti-recurrence of RCC after operation is expected. In this study the purifed HCC cells and autologuous activated B cells from HCC tissue and autologous spleen were used. Hybrid cells were generated by fusion of human hepatocellular carcinoma cells (HCC) with autologous activated B cells of HCC patients and were irradiated by 60Co. Hybrid cells were determined by flow cytometry and the fused proportions were 66.84%,74.43% and 76.55%, respectively. The hybrid cells were able to express both gp75 molecules from human HCC cells ,and MHC-I,MHC-II and B7 from B cells. 60Co-radiated fusion cells lose proliferative capacity ,but they have no apparent changes in immunogenicity-related phenotype MHC-I,MHC-II and B7 in subsequent 3 d. Asepsis, pyrogen2and endotoxin accord with the criterion of the national preparation and testing of bioproduction. Remnant PEG,glycerol and gentamicin were tested. The results were: PEG was not detected, remnant glycerol and gentamicin were less than 0.02% and 1mg/i, respectively. RemnantPEG, glycerol and gentamicin in the sample are much less than the harmful dose in vivo. These results suggest the safty and efficiency of the hybrid cell vaccine.