摘要
目的 证实 NK细胞存在与 T细胞类似的 Th1/Th2两类亚群。 方法 从健康人外周血分离纯化 NK细胞，利用促漂移和 RT-PCR的技术，检测外周血静止的 NK细胞及在不同极化条件下培养的 NK细胞表达Ⅰ和Ⅱ类细胞因子的情况。 结果 采用补体裂解法＋单抗铺皿法分离纯化 15例健康人外周血 NK细胞， RT-PCR方法检测 NK细胞表达的两类因子，发现外周血静止 NK细胞中Ⅰ类细胞因子的表达较强，主要是 IFNγ，Ⅱ类细胞因子中主要是 IL-10、 IL-13。在促 Th1极化条件下， NK细胞中表达的Ⅰ类细胞因子 IFNγ 明显增强，Ⅱ类细胞因子表达减弱，而在促 Th2极化状态下， NK细胞表达的 IFNγ水平下降，Ⅱ类细胞因子的表达水平升高。静止和Ⅱ类细胞因子状态下 NK细胞均表达极低水平或不表达 IL-4。 结论 NK细胞根据其产生的细胞因子可以分化为两个亚群，暂命名为 NKh1和 NKh2。外周血中静止的 NK细胞主要以 NKh1表型为主，两个亚群的 NK细胞均表达较高水平的 IFNγ，这与 NK细胞的杀伤功能是密切相关的。
Objective To verify the presence of functional subsets of natural killer cells based on the cytokine production. Methods NK cells were purified and cultured in complete RPMI1640 medium in the presence of either IFNγ＋ anti-IL-4(classical Th1 polarization) or IL-4＋ anti-IFNγ (classical Th2 polarization) for three days, and then were collected and detected for typeⅠ /typeⅡ cytokines by RT-PCR method. Results NK cells were purified from 15 healthy donors,over 70％ purity of NK cells were determined by flow cytometry. NK cells in peripheral blood expressed high level of typeⅠ cytokines,mainly IFNγ , but low level of typeⅡ cytokines such as IL-10 and IL-13, IL-4 was not produced by NK cells. Cells cultured in IFNγ＋ anti-IL-4 condition exhibited significantly increased level of IFNγ ,unchanged IL-2,and decreased typeⅡ cytokines. Cells grew in IL-4＋ anti-IFNγ condition exhibited increased IL-10 and IL-13, and decreased IFNγ expressions. Conclusions Based on the cytokine production,NK cells may be divided into two functional subsets in the same manner as that of T lymphocytes(e.g. Th1/Th2):NKh1 and NKh2. The biological characterization and phenotypic marker are under investigate.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2001年第2期132-136,共5页
Acta Academiae Medicinae Sinicae
基金
国家自然科学基金! (39870729
39970694
39970828)&&
