People From Hepatoma Cell Lysates Pulsed Dendritic Cell Vaccine To Prevent Recurrence And Metastasis Of Hepatocellular Carcinoma After Surgery

Objective : To study the method of induction, proliferation and biological character of dendritic cells (DCs) derived from peripheral blood mononuclear cell of hepatocellular carcinoma (HCC) patients, and further to prepare the hepatoma cell lysates pulsed DC vaccines in vitro. It will provide the experimental foundation for clinical immunotherapy of the hepatoma cell lysates pulsed DC vaccines. Method: 1.The induction and biological character of DCs derived from peripheral blood mononuclear cell of HCC patients. DCs isolated from peripheral blood mononuclear cell of HCC patients were cultured and proliferated in vitro by using recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) and interleukin-4 (rhIL-4),and then were analyzed on morphology under optics microscope; the phenotypes of DCs are detected by FACS analysis including specific CD1a, and function-related molecules CD40, CD86, HLA-DR; the assay of mixed lymphocyte reaction(MLR)is performed by using 3H-TdR penetration method to observe the ability of DC stimulating lymphocytes proliferation. 2. Preparation and identification of the hepatoma cell lysates pulsed dendritic cell vaccines 2.1 Hepatoma cells were isolated from resectable hepatoma tissue in patients with HCC, and then the hepatoma cell lysates were prepared from hepatoma cells. 2.2 DCs were isolated from peripheral blood mononuclear cell of HCC patients in a similar way as above,and then were pulsed with autologous hepatoma cell lysates. 2.3 The hepatoma cell lysates pulsed dendritic cell vaccines were identified through morphology, FACS and MLR. Result: 1. The identification of DCs and its biological character Typical DCs could be induced when peripheral blood mononuclear cell of HCC patients were cultured in medium with rhGM-CSF and rhIL- 4 for 7-14 days. It showed typical morphological of dendritic cell under optics microscope;It highly expressed the specific cellular phenotype CD1a and other functional-related molecules such as CD40, CD86, HLA-DR; Theassay of MLR showed that DCs could stimulate strong T lymphocytes proliferation. 2. Preparation and identification of the hepatoma cell lysates pulsed dendritic cell vaccines 2.1 Hepatoma cells and their lysates were prepared from resectable hepatoma tissue in patients with HCC. 2.2 We succeeded in preparing the hepatoma cell lysates pulsed dendritic cell vaccines. 2.3 The hepatoma cell lysates pulsed DC vaccines Led to up-regulation of CD1a, CD40, CD86 and HLA-DR. Autologous lymphocytes proliferation was markedly enhanced in hepatoma cell lysates pulsed DC vaccines group than that in unplused DCs group, hepatoma cell lysates group and control group (P<0.01). Conclusion: 1. Typical DCs could be induced when peripheral blood mononuclear cell of HCC patients were cultured in medium with rhGM-CSF and rhIL- 4 for 7-14 days. It had potent antigen presenting functions. 2. We succeeded in preparing the hepatoma cell lysates pulsed dendritic cell vaccines. 3. The hepatoma cell lysates pulsed dendritic cell vaccines enhanced the antigen-presenting functions of DCs, which provided the experimental foundation for clinical immunotherapy of the hepatoma cell lysates pulsedDC vaccines. Objective:To investigate T cell-mediated antitumor effects in vitroelicited by dendritic cells pulsed with tumor cell lysates in patients withhepatocellular carcinoma. It will offered a new opportunity for clinicalimmunotheray of HCC. Method: 1. DCs were isolated from peripheral blood mononuclear cell of HCCpatients in a similar way as above,and then were pulsed with autologoushepatoma cell lysates. 2. The phenotypes of the hepatoma cell lysates pulsed dendritic cellvaccines were assayed by FACS . 3. The concentration of IFN-γ and IL-12 were measured in culturesupernatants by ELISA. 4. The ability of DCs pulsed with hepatoma cell lysates to stimulate proliferation of autologous T lymphocytes was tested by 3H-TdR penetration method. 5. The cytotoxicity of CTL stimulated by hepatoma cell lysates pulsed DC vac...