| Objective:To prepare hepatoma carcinoma cells HepG-2 antigen in vaccine and to investigate the antitumor effects of hepatoma carcinoma cells HepG-2 antigen in vaccine accompanied by CpG cantaining oligodeoxynucleotide (CpG ODN) adjuvant in vitro.Methods:Peripheral blood monouclear cells was co-cultuered in 96 well cell culture plates with hepatoma carcinoma cells HepG-2 antigen or CpG 2006 or both hepatoma carcinoma cells HepG-2 antigen and CpG 2006,to detecte the activations of the peripheral blood monouclear cells and cyto toxincity of the peripheral blood monouclear cells to hepatoma carcinoma cells HepG-2 in virto,there are six groups in this experiment:control group:on antigen and CpG ODN added;antigen group:on CpG ODN added,numbers of PBMC and numbers of hepatoma carcinoma cells HepG-2 as antigen is 20:1;CpG ODN group: no antigen added, the CpG ODN's final concentration added is 1μl.ml-1;CpG ODN and antigen group 1:antigen added as the antigen group, CpG ODN added as the CpG ODN group; CpG ODN and antigen group 2:antigen added as theantigen group, the CpG ODN's final concentration added is 2μl.ml-1; CpG ODN and antigen group 3:antigen added as the antigen group, the CpG ODN's final concentration added is 5μl.ml-1.To regulate all groups to 200μl with RPMI 1640 cell culture fluid. each group has 3 wells.all groups are co-cultured three days .then to detect IFN-gama's concentration of each well's supernatant and co-culture the PBMC and hepatoma carcinoma cells HepG-2 for 48 hours.after 48hours ,to detect the LDH concentration of each mixed cell's supernatant to observe the degree of the injure of the PBMC to hepatoma carcinoma cells HepG-2.Results:At present,it is presumed that the carcimoma immunotherapy has intimate relationship with Th1 type immunological reaction ,and Th1 type immunological reaction has intimate relationship with IFN-gama,so IFN-gama is detected .to compare the control group,antigen group , CpG ODN group , antigen and CpG ODN group 1,find that PBMC of all group secrete IFN-gamma , the capability of stimulating PBMC to secrete IFN-gamma than the other groups as follows (from higher to lower ): antigen and CpG ODN group,CpG ODN group ,antigengroup,control group,the difference is obvious (p<0.05,Figure 1).illustrate that both antigen and CpGODN has higher capability to stimulate PBMC to generate Th1 type immunological reaction .to compare the antigen and CpG ODN group 1, antigen and CpG ODN group2, antigen and CpG ODN group 3,find that while the antigen is invariable ,between 0-5μg.ml-1,the capability to stimulate PBMC to secrete IFN-gamma is higher when the concentration of CpG ODN is higher,the difference is obvious (p<0.05). To compare the control group,antigen group , CpG ODN group , antigen and CpG ODN group 1's injury to the hepatoma carcinoma cells HepG-2 ,find that the capability of the PBMC to injury the heaptoma carcinoma cells HepG-2 as follows(from higher to lower ): antigen and CpG ODN group,CpG ODN group ,antigen group,control group,the difference is obvious (p<0.05). illustrate that both antigen and CpGODN has higher capability to stimulate PBMC to generate the capability to injury carcinoma cells .to compare the antigen and CpG ODN group 1, antigen and CpG ODN group2, antigen and CpG ODN group 3,find that while the antigen is invariable ,between 0-5μg.ml-1,the capability to stimulate PBMC to generate the capability to injury ishigher when the concentration of CpG ODN is higher,the difference is obvious.Conclusions:Hepatoma carcinoma cell antigen or CpG ODN alone can enhance the anti-tumur activity of PBMC , but the enhancement of the combination of hepatoma carcinoma and CpG ODN to the anti-tumur activity of PBMC is unique . And while CpG ODN combining with hepatoma carcinoma cell antigen stimulate the anti-tumur activity of PBMC , and the concentration of CpG ODN lower than 5μg.ml-1 , the enhancement of the antitumur activity of PBMC has an positive relevance with the concentration of CpG ODN...
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