STUDIES ON PHENOTYPE OF CIK AND ITS BIOLOGICAL ACTIVITIES ON TUMOR CELLS / 解放军医学杂志

ABSTRACT

Objective To generate cytokine induced killer (CIK) by inducing lymphocytes cells from peripheral blood in vitro , and to study the phenotype and biological activities of CIK. Methods Lymphocytes cells were isolated fresh from peripheral blood of healthy donors by Ficoll Hypaque density centrifugation, and the cells obtained were resuspended in RPMI medium consisting of 10% fetal calf serum at 37℃, 5%CO 2 for 2h. On day 0, the nonadherent cells were activated with IFN ? (1 000U/ml) and the following days were stimulated with CD3mAb (3 75?g/ml) and rhIL 2 (600U/ml). Phenotype of CIK were analysed by FACS. The proliferation of CIK was tested using 3 H Thymidine, and the killing experiment using MTT tests. Results The proliferation peak of CIK was at day 20, and declined at day 30. The percentages of the cytotoxicity of CIK cells for stomach cancer cell line MGC803 and liver cancer cell line Bel7402 was 65 45%?5 68% and 68 37%?3 93% respectively. CIK cells expressed the phenotypes of CD3CD56, CD3, CD54, CD28CD54, CD11a, HLA DR and CD28. Conclusion IFN ?, CD3mAb and rhIL 2 can induce strong proliferation of peripheral lymphocytes to produce CIK cells expressing the phenotype of CD3CD56, CD3, CD54, CD28CD54, CD11a, HLA DR, CD28, which have strong cytotoxicity on tumor cells.