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目的:建立一种检测小鼠Ⅱ型NKT细胞的方法。方法:将硫酸脑苷酯(sulfatide)与生物素连接的CD1d单体按摩尔比3∶1混合室温静置过夜,之后加入PE标记的链酶亲和素,室温孵育4 h制成sulfatide/CD1d四聚体。然后应用流式细胞术(FCM)检测正常小鼠脾脏及肺脏单个核细胞中Ⅱ型NKT细胞的百分比(TCR-β+sulfatide/CDld四聚体+),以及小鼠脾细胞经sulfatide刺激后单个核细胞中的Ⅱ型NKT细胞的百分比。结果:正常小鼠脾脏及肺脏单个核细胞中Ⅱ型NKT细胞的比例分别为(0.875±0.096)%和(1.175±0.263)%;小鼠脾细胞经sulfatide刺激后的Ⅱ型NKT细胞的比例为(2.75±0.603)%,与正常相比明显增加(P<0.01)。结论:硫酸脑苷酯负载的CDld四聚体是检测小鼠Ⅱ型NKT细胞的有效方法。
Objective: To establish a method for detecting type Ⅱ NKT cells in mice. Methods: The CD1d monomers with sulfatide and biotin were mixed at the molar ratio of 3: 1 overnight at room temperature. Then PE labeled streptavidin was added and incubated for 4 h at room temperature to prepare sulfatide / CD1d Tetramers. The percentage of type II NKT cells (TCR-β + sulfatide / CDld tetramer +) in spleen and lung mononuclear cells of normal mice was detected by flow cytometry (FCM) Percentage of type II NKT cells in nucleated cells. Results: The percentage of type Ⅱ NKT cells in spleen and lung of normal mice were (0.875 ± 0.096)% and (1.175 ± 0.263)%, respectively. The percentage of type Ⅱ NKT cells in spleen cells stimulated by sulfatide was (2.75 ± 0.603)%, significantly increased compared with the normal (P <0.01). Conclusion: The cerebroside-loaded CDld tetramer is an effective method for the detection of type II NKT cells in mice.