| ObjectiveThe transcatheter arterial chemoembolization(TACE) is the main treatment for people with inoperable primary hepatocellular carcinoma(HCC), it can prolong people's survival, but there often showed uncomplete necrosis in lesions and immune function damage in patients after TACE. In recent years, with the development of cellular immunology and molecular biology, cellular immunotherapy has been applicatied in HCC,and has achieved good results, in which DC and CIK cell-based anti-tumor immunotherapy has became a'hot research at home and abroad for its unique advantages.Dendritic cells (DC) is regarded as a powerful antigen-presenting cell (APC), the cytokine-induced killer cells (CIK) is a novel immune active cells, both with strong anti-tumor activity of T lymphocytes and restricted killing tumor characteristics of major histocompatibility complex (MHC) of natural killer cells(NK) cells. DC and CIK is an important part of tumor immunotherapy, the former identify pathogen and activate the acquired immune system, the later can kill tumor cells through cytotoxicity and secretion of cytokines. When CIK cells and DC cells were co-cultured,it not only improves the ability of CIK cells proliferation and cytotoxicity, but also can significantly improve the DC cells'function.In this study, we co-cultured the HepG2-antigen pulsed autologous DC cells with CIK cells, and no-antigen pulsed autologous DC cells with CIK cells, respectively.We observated the variation of phenotype,proliferative activity and cytotoxicity of the culture. The patients were treated with Ag-DC-CIK after TACE therapy,immune function,quality of life,clinical efficacy,AFP,HBV-DNA and adverse were observed and it provided the basis for clinical bioimmunotherapy of HCC.Methods1. DC cells and CIK cells were generated from culturing peripheral blood mononuclear cells(PBMC), Phenotype of these effector cells were identified by flow cytometry analysis. HepG2 soluble antigen was prepared.We co-cultured the HepG2-antigen pulsed DC with CIK(Ag-DC-CIK) and non HepG2-antigen pulsed DC with CIK.The proliferation and the killing activity of cells were observed dynamicly.2. A total of 41 patients with HCC were randomly divided into two groups:20 patients in treatment group, received TACE first then were given Ag-DC-CIK treatment,21 patients in control group,only received TACE therapy.3. The level of T cell subsets,CD4+CD25+ regulatory T cell and NK cells were detected by flow cytometry analysis before and after the treatment. The quality of life,tumor,AFPand HBV-DNA changes,and side reactions were aslo analyzed.Results1. CIK, DC-CIK and Ag-DC-CIK began proliferating on day 3, and into the fast-breeder period on day 12. The proliferation of Ag-DC-CIK group faster than CIK and DC-CIK group (p<0.05).2. Three groups of cells expressed CD3+CD8+, CD3+CD56+double positive cells, the percentage of CD3+CD8+, CD3+CD56+double positive cells of three groups was increasing with the incubation time, the percentage of CD3+CD8+, CD3+CD56+ double positive cells in Ag-DC-CIK group were significantly higher than CIK and DC-CIK group that of the same culture time, (p<0.05).3. The killing activity of antigen pulsed DC-CIK was the strongest in the E:T range of 20:1,and it had significant difference compared with CIKand DC-CIK group (P<0.05). 4. In the treatment group, the percentage of CD3+,CD4+,CD16+CD56+ effect cells and the proportion of CD4+/CD8+ increased and the percentage of CD4+CD25+,CD8+ effect cells decreased significantly after the treament(p<0.05).5. The complete response rate(CR),partial response rate(CR)and response rate(RR)of treatment group were higher than the control group, but there were no significant difference (P> 0.05). The general condition of patients in treatment group were significantly improved compared with the control group(p<0.05).6. There is 69.2% of patients in the the treatment group, AFP down to 400ug/L from more than 400ug/L, significantly higher than the control group(p<0.05).7. There were 60% patients whose HBV-DNA become negative in treatment group, significantly higher than the control group(p<0.01)8. In the course of treatment, two cases in treatment group appeared chills fever, usually around 38, all can be alleviated after symptomatic treatment.We didn't observed other serious adverse reactions, such as allergy, shock, myelosuppression, liver and kidney failure.Conclusions1. The main effector cells of co-cultuerd antigen-pulsed DC and CIK are the CD3+CD8+ and CD3+CD56+ double positive cells; antigen-pulsed DC cells can enhance the proliferation activity of CIK cells; antigen-pulsed DC co-cultured with CIK can achieved strongest killing activity.2. After treatment of the Ag-DC-CIK cells, the ordinary condition of patients with advanced hepatocellular carcinoma had been significantly improved, the imbalance of T cell subsets has been improved to some extent, tumor has been effectively controlled, the quality of life has been improved obviously, and no significant adverse reactions were obsered. |
PDF Full Text Request