| [Objection] : To estabilish the method of inducing the dendritic cells from human peripheral blood monocytes(PBMC) and cord blood (CB)CD34+ cells, and study their function on the anti-tumor immunity. [Methods] Mononuclear cells were separated from peripheral blood on Ficoll hypague gradient, monocytes were obtained by adherent treatment and were cultured in the midia with GM-CSF, IL-4 and TNF-α. CD34+cells were isolated from umbilical cord blood by Magnetic Activated Cell Sorting(MACS) system. CD34+ cells were induced to DC with cytokines:GM-CSF, IL-4, TNF-α, SCF, FL, EPO. The morphology of DCs were observed under the electronic microscope and optical microscope.The phenotype of DCs were analysed by Flow Cytometry . T lymphcytes proliferation ability was tested with allo mixed lymphocyte react ion (allo-MLR). Abilities of cytokines TNF-a, IL-lp, IFN-y and CDSmAb to stimulate DC to mature were also analysed and the abilities of the mature DCs to stimulate CTL to kill the leukaemia cell line K562 were tested. [Results] DCs wre obtained from PBMC by using GM-CSF, IL-4, TNF-α , and also can be induced from CB-CD34+ by using GM-CSF, IL-4, TNF-α , SCF, FL, TPO. Cord Blood (CB)CD34+ cells had strong expansion ability in this culture condition. The expensive fold reach to 115.7+ 24.5 times after being induced for two weeks. PBDCs expressed CD1a47. 6%, CD14 5. 2%, CD80 8. 87%, CD83 25. 9%, CD86 59. 5 and HIA-DR 97. 3% in day 7. CBDCs expressed CD34 2. 6,CDla 31.3%,CD14 16. 0%, CD80 13. 9%, CD83 14. 6%, CD86 18. 1% and HLA-DR 71. 5% in day 14. The DCs from different sources could stimulate allogeneic lymphocytes proliferation reaction. TNF-α, IL-1β and IFN-r could stimulate DCs' maturation and DCs could stimulate CTL to kill the tumor cells effectively after loaded with the tumor antigens. [Conclusion]DCs could be obtained from both the peripheral blood monocytes and the cord blood CD34+ cells in vitro and they had the same phenotypes and functions on stimulating the lymphocyte proliferation.
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