Experimental Study On The Anti-tumor Role Of Natural Killer T Cells With Dendritic Cells

[Objective] To establish purified highly natural killer T cell in vitro and applie flow cytometry to carry on the analysis to its phenotype. On this basis, to study the Anti-tumor Role and mechanism of natural killer T cells and dendritic cells.[Results] Peripheral Blood was collected under sterile condition and the Peripheral Blood Mononuclear Cells (PBMC) were Separated by centrifugal in density gradient. PBMC were cultured with IL-4+GM-CSF+TNF-αand cell phenotype was analyzed with CD1a,CD80,CD86,CD83 antibody by using indirect immunofluorescence assays. T cells from the donor cultured with nature dentritic cells in the tweleve days can induce the proliferation of NKT, and cell phenotype was analyzed with CD3,CD161 antibody by using indirect immunofluorescence assays. strongestcytotoxicity was observed to all tumor cell lines AGZY 83-a and Anip973.[Results] PBMC Separated from the human Peripheral Blood were cultured for 10 days, we can see DCs of representative configuration structure under light microscope, cell phenotype was analyzed with CD1a,CD80,CD86,CD83 antibody by using indirect immunofluorescence assays. The experiment demonstrated that, DC can succeed induces proliferation of the NKT cell, the DC low density stimulation group is control group's 0.82times in the NKT multiplication degree; the DC median density stimulation group is control group's 1.49 times in the NKT multiplication degree; the DC high density stimulation group is control group's 2.51times in the NKT multiplication degree. Indicated that, along with proportion enlargement of the dentritic cells and NKT cell, NKT cell multiplication ability strengthens gradually, simultaneously the result also showed, regardless of what kind of density DC, its ability in stimulating the NKT cell multiplication strongly always in the cell factor stimulation group. DC raises which using this testing method may effective activate normal healthy person circumference blood NKT cell and promotes its multiplication, and may strengthen the NKT cell the cytotoxin function., When effect: target is 5:1, the suppression rate of the NKT cell to the AGZY 83-a tumor cell is 16.9±12.4%, to Anip 973 tumor cell s is 19. 6±13.8%; When effect: target is 10:1, the suppression rate of the NKT cell to the AGZY 83-a tumor cell is 38.8±9.6%, to Anip 973 tumor is 52.8±9.4%; When effect: target is 20:1, the suppression rate of the NKT cell to the AGZY 83-a tumor cell is71. 6±11. 2%, to Anip 973 tumor cell is 80. 3±8. 0%. Indicated that, anti-tumor activeness of NKT cell increases along with the effect and target proportion unceasing ascension, similarly explained, killing activeness of NKT cell to Anip 973 tumor cell is stronger than AGZY 83-a tumor cell. DCs has the ability to expand massively increases the NKT cell in vitro, and may activate the NKT cell and play the anti-tumor role. For further developed the NKT cell the foundation and the clinical practice research has laid the foundation andAlso will open a new effective way for the tumor and the tumor shift diagnosis and the prevention.[Conclusion] 1. Peripheral Blood Mononuclear Cells (PBMC) were produced DC with IL-4 + GM-CSF + TNF-α. 2. DC can succeed induces proliferation of the NKT cell, and activation of NKT cell can play the anti-tumor role. 3. The NKT cell is stronger to the Anip973 killing activeness and may suppress the tumor shift...