IFN-γ诱导THP-1源巨噬细胞活化的条件优化

Optimization of IFN-γ-Induced THP-1 Macrophage Activation

目的摸索利用IFN-γ诱导THP-1源巨噬细胞活化的方法，筛选用于检测巨噬细胞活化状态的最佳细胞因子和最佳检测时间。方法PMA（佛波酯）诱导THP-1分化成为巨噬细胞，两次加入IFN-γ诱导巨噬细胞活化（两步法），ELISA法检测不同浓度和不同刺激时间培养上清中细胞因子TNF-γ、IL-10、IL-12、IL-23的水平。结果IFN-γ诱导的巨噬细胞可大量分泌TNF-a，分泌量大于1000ng／mL，而IL-10、IL-12、IL-23分泌量少或未见明显分泌；两次IFN—y诱导巨噬细胞活化的最低浓度均为2．5ng／mL，初次最佳刺激时间为8～16h，再次最佳刺激6～18h。结论IFN—y可诱导巨噬细胞活化，两次IFN-γ刺激的效果比单次刺激效果好，TNF-γ可作为巨噬细胞活化的理想检测指标。

Objective To find out the optimal cytokines and detection time when the activation state of macrophages derived from THP-1 induced by IFN-T was detected. Methods THP-1 cells which differentiated into macrophages induced by phorbol ester （PMA）, IFN-y were added to induce the activation of macrophages for twice. ELISA was used to detect the concentration of TNF-a, IL-10, IL-12 and IL-23 in cultured supematant in different stimulation concentration and stimulation time. Results The macrophages induced by IFN-y could secrete a mass of TNF-a which could increase to over 1 000 ng/mL. At the same time, there were no obvious concentration changes of IL-10, IL-12 and IL-23. The minimum concentration of IFN-y to induce macrophages activation was 2.5 ng/mL for each time. The optimal first stimulation time was 8 to 16 hours while the optimal second stimulation time was 6 to 18 hours. Conclusions Macrophages can be induced by IFN-y. Stimulation by IFN-y for twice is more effective than once, therefore, it can be an ideal detection index for macrophage activation.