Activity Of CTL Induced By DC Transfected By Recombinant Adeno-associated Virus Mediatedcarcinoembryonic Antigen(CEA) Gene

Objective:To investigate the surface marker、CEA expression of DC and the antitumor efficiency of the special cytotoxic T lymphocytes (CTLs) activated by dendritic cells of colorectal cancer patients transfected by adeno-associated virus mediated CEA gene by flow cytometry,we can arrive the conclusion that this method can lift the surface marker、CEA expression of DC and the antitumor efficiency of the special cytotoxic T lymphocytes (CTLs).Methods:Using density gradient centrifugation, the mononuclear cells were isolated from peripheral blood of colorectal cancer patients,then they were cultured in AIM medium containing rhGM-CSF、rhIL-4and rhTNF-a to induce DC. It was divided into experimental and control groups,experimental group was added recombinant adeno-associated virus mediated CEA gene to transfect DC, while control group was adding the same amount of PBS. The surface marker and CEA expression of DC、specific killing of CTL cells were detected by flow cytometry. Statistical analysis was performed by using the software SPSS16.0.Results:1DC has an irregular change of shape on the second day,then its cell volume is gradually becoming bigger and emerging various Synaptic after infusing of rhTNF-a on the fourth day,which compared with normal DC, it is suggesting that the rAAV transfected virus does not affect the growth of DC.2the expression rates of CD40、CD80、CD86in the experimental group is (81.95±1.45)%(80.10±1.03)%、(58.93±1.84)%,while control group is (72.27±1.59)%、(70.59±1.04)%、(48.66±1.97)%,respectively. The difference was statistically significant, P<0.05.3The expression rate of CEA in the experimental group is (59.04±1.68)%,while control group is (46.03±1.23)%.The difference was statistically significant, P<0.05.4The effector cells to target cells is12.5:1、25:1、50:1、100:1,specific killing of CTL is (12.65±0.30)%、(25.77±2.53)%、(43.36±0.83)%and(61.03±1.83)%,repectively,significantly higher than those of control group((6.59±0.21)%、(8.35±1.12)%、(12.79±0.23)%and (23.35±0.69)%. The difference was statistically significant, P<0.05.Conclusion1The rAAV transfected virus does not affect the growth of DC.2The expression rates of CD40、CD80、CD86in the experimental group is higher than those of control group.3The expression rates of CEA in the experimental group is higher than those of control group.4The special cytotoxic T lymphocytes (CTLs) activated by dendritic cells (DC) transfected by recombinant adeno-associated virus mediated CEA gene have higher antitumor activity.