| Objective:With human peripheral blood mononuclear cells used as effector cells, and chronic myelogenous leukemia K562 cell line as target cells, this experiment is aimed at studying synergistic effect on human peripheral blood mononuclear cells (PBMC) of PPS combined with IL-15, as well as further clarifying the effect on its killing activity and probable function to fight back the chronic myeloid leukemia K562 cells.Methods:Firstly, this experiment uses Ficoll density gradient method to separate PBMC. Secondly it applies respectively IL-2, IL-15, PPS, PPS+IL-2 and PPS+IL-15 to stimulate the proliferation of peripheral blood mononuclear cells, in order to acquire the effector cells. Thirdly, it need to detect activated Immunophenorype of T cells and NK cells in PBMC by flow cytometry. Besides, this experiment uses double antibody sandwich ELISA method to detect the level of IFN-y and TNF-α in cell culture supernatant, then after extracting chronic myeloid leukemia K562 cell line in logarithmic growth phase and cultivating with effector cells, it detect the cytotoxic effect of effector cells to target cells by MTT colorimetry.Results:Compare with the control group, the combine of PPS and IL-15 can stimulate the proliferation of the PBMC as well as raise the level of IFN-γ and TNF-α in effector cell culture supernatant; and the effector cells stimulated by PPS and IL-15 have conspicuous killing activity to chronic myeloid leukemia K562 cells. All of the above conclusion are statistically significant (P <0.05).Conclusion:PPS and IL-15 can stimulate the proliferation of T cells, NK cells in PBMC. It has synergistic effect on promoting the proliferation of NK cells. It also raise the level of cytokines IFN-γ and TNF-α in effector cell culture supernatant, which have obvious cytotoxicity to chronic myeloid leukemia K562 cells, and its cell killing mechanism may be related to the level improvement of cytokines IFN-γ and TNF-α in effector cell culture supernatant. |
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