The Research Of The Factors In The Expansion In Vitro And The Killing Activity Of The CIK And NK Cells Derived From Cancer Patients' Peripheral Blood

Objective:To study the factors of cancer patients on the expansion ability of their own CIK and NK cells in vitro and detect the antitumor activity of the CIK and NK cells. In order to provide a theroretical basis for the treatment of CIK and NK cells against the malignant tumors.Materials and methods:The first part, we analyzed the patiens with malignant tumors who had received CIK and NK autoimmune therapy, to explore the effects of age, gender, tumor type, stage and the distribution of the lymphocyte sub-group on the expansion of CIK and NK cells. we counted the number of the cells everday and observed the morphological changes before and after culture by inverted microscope, and the changes of CD3+CD56+ and CD3-CD56+ cells,and we harvest the CIK and NK cells at the 14th days. The second part, we detected the secretion level in the supernatant of the culture system by ELISA test kit. And we detect the killing activity of CIK and Nk against K562 cell line by MTT and Calcein-AM/PI fluorescence staining method.Results:1. The expansion of the patients'CIK and NK cells was lower than the normal,but had no relevance with age, gender and the turmor type. However, the stage and the distribution of the lymphocyte sub-group were the factors in the expansion of CIK and NK cells. The expansion of the CIK and NK cells derived from stageⅢ-Ⅳpatients Was lower than the CIK and NK cells derived from stageⅠ-Ⅱpatients. After induction and expansion, the percentage of NK cells had a positive correlation with the percentage of NK cells before culture, and had a negtive correlation with the percentage of CD8+ cells before culture.2. After induction and expansion in vitro for 14 days,the percentage of CIK and NK cells raised to 30.78±15.29% and 58.32±10.13%, from 2.03±0.17% and 11.32±3.42%, respectively. The secretion level of IFN-γraised to 10.08±0.44pg/mL,14.76±0.42pg/mL,were lower than the normal CIK and NK cells.3. Before culture, the killing activity of PBMC from the patients and the normal was simililar at any effectors to targeted ratio. But after induction and expansion,the killing activity of CIK and NK cells against K562 cell line enhanced greatly compared with the PBMC. With the increase in effectors to targeted cells, the antitumor activity also increased at some degree. And the combining of CIK and NK cells had the most cytotoxicity,When the effectors to targeted cells was 25:1, the CIK and NK cells could kill the 80% targeted cells.Conclusion:First, the expansion of the CIK and NK cells derived from cancer patients was lower than the normal. But had no relevance with the patients' age, gender and turmor type. But, the stage and the distribution of the lymphocyte sub-group were the factors in the expansion of CIK and NK cells in vitro. Second, we can obtain a large number of CIK and NK cells by induction and expansion in vitro, and the secretion level of IFN-γand the killing activity against K562 cell line were enhanced, and the combining of CIK and NK cells had the most cytotoxicity. Consequently, CIK and NK cells can be used to treat malignant patients in clinical as a new type of adoptive immuno effector cells.