The Effects Of CIK Cells On PD-1 And Lymphocyte Subpopulation In CIK Adoptive Immunotherapy

Objective: 1.To analysis if PD-1 pathway is involved in the anti-tumor process of CIK cells by investingating the influence of CIK cells induction to PD-1. 2.Inquiry the variation of lymphocyte subpopulation in CIK cells which obtained after immunotherapy to determine the CIK treatment has any impacts on the immune cells of cancer patients.Methods: 1.36 pathologically confirmed malignant tumor patients who were voluntary committed for CIK adoptive immunotherapy and had not previously recieved similar treatment. They were screened as experimental group. We also randomly selected 14 normal adults who had no tumor or immune diseases as control group. 2. Before the first and the third cycle of CIK treatment, we extracted 50-100 ml peripheral blood of patients through elbow vain and 2ml of them was used for flow cytometry testing. The rest blood was induced with other cytokines in vitro and transformed into CIK cells after cultivation. Collected 2ml CIK cells for flow cytometry testing and the rest was instilled to the patients. 3. We use flow cytometry to compare the different expression levels of PD-1 before and after immunotherapy or induction respectively, to analysis CIK cell lymphocyte subpopulation expression level differences before and after CIK treatment, to analysis the relationship between the expression of PD-1, CD3+ CD56+, CD4+ / CD8+ expression and treatment,clinical stage before and after CIK treatment.Results: 1. The expression of PD-1 after induction was higher than those before, which was more obvious after immunetherapy(P<0.05). After CIK treatment,PD-1 in peripheral blood was lower while that in CIK cells was higher, but the differences above had no statistical significance(P>0.05). The PD-1 expression levels in peripheral blood has no statistical difference between healthy people and cancer patients before CIK treatment(P>0.05). 2.Expression of CD3+CD56+ in CIK cells was higher after immunotherapy and the difference had statistical significance(P<0.05). CD3+, CD4+, percentage of lymphocytes and the ratio of CD4+/CD8+ all elevated while CD8+ decreased after CIK treatment. Differences above had no statistical significance(P>0.05). 3.Initial expression levels of PD-1 and CD4+/CD8+ were lower in patients of stage III~IV than those in patients of stage I~II while initial expression of CD3+CD56+ was higher in patients of stage III~IV than those in patients of stage?~?, but the differences had no statistical significance(P>0.05). After CIK treatment, PD-1 expression level in patients of stage III~IV was elevated while in patients of I-II was reduced, but the CD3+CD56+, CD4+/ CD8+ expression levels were elevated in patients of all sages. Rangeability of CD3+CD56+ in CIK cells and PD-1 in peripheral blood was wider in patients of stage I~II while that of PD-1 and CD4+/ CD8+ in CIK cells was wider in patients of stage III~IV, the differences above had no statistical significance(P>0.05). 4. The expression of PD-1, CD3+CD56+ and CD4+/CD8+ changed after CIK treatment, but the variation was not influencd by surgery, chemotherapy, radiotherapy and clinical stage.Conclusion: 1. The induction and cultivation process of CIK cells in vitro can up-regulate the expression of PD-1. 2. CIK adoptive immunotherapy can suppress the expression of PD-1 in early stage cancer patients. 3.CIK adoptive immunotherapy can improve the proliferation capacity of CD3+CD56+ precursor cells in vitro.