J Fang1, Z Tian, C Zhang. 1. Shandong Cancer Biotherapy Center, Basic Medical Institute, Shandong Academy of Medical Sciences, Jinan 250062, China.
Abstract
OBJECTIVE: This work was to do preliminary study on the characteristics and preparation of rhIL-15 induced adherent human natural killer cells (A-NK). METHODS: Natural killer cells (NK cells) were first separated by centrifugation on Ficoll-Hypaque gradients, plastic adherence and nylon wool column adherence. Then, they were further purified by Percoll discontinuous density gradient centrifugation and T cell panning. Fluorescence-activated cell scan (FACScan) was applied to evaluate the natural killer cells and assess their degree of purification. Then, the purified NKs were incubated in the presence of rhIL-2 (6,000 U/ml) or rhIL-15 (6,000 U/ml) and changed into adherent NK cells. In the next step, the adherent kinetics, proliferation and cytotoxicity of A-NKs obtained by two different cytokines were analyzed by cell counting, MTT and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The adherent percentage, cytotoxity and proliferation of A-NKs generated in rhIL-15 culture were higher than those in rhIL-2. CONCLUSION: IL-15 is a better stimulator than IL-2 to induce adherent natural killer cells.
OBJECTIVE: This work was to do preliminary study on the characteristics and preparation of rhIL-15 induced adherent human natural killer cells (A-NK). METHODS: Natural killer cells (NK cells) were first separated by centrifugation on Ficoll-Hypaque gradients, plastic adherence and nylon wool column adherence. Then, they were further purified by Percoll discontinuous density gradient centrifugation and T cell panning. Fluorescence-activated cell scan (FACScan) was applied to evaluate the natural killer cells and assess their degree of purification. Then, the purified NKs were incubated in the presence of rhIL-2 (6,000 U/ml) or rhIL-15 (6,000 U/ml) and changed into adherent NK cells. In the next step, the adherent kinetics, proliferation and cytotoxicity of A-NKs obtained by two different cytokines were analyzed by cell counting, MTT and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The adherent percentage, cytotoxity and proliferation of A-NKs generated in rhIL-15 culture were higher than those in rhIL-2. CONCLUSION:IL-15 is a better stimulator than IL-2 to induce adherent natural killer cells.