Sulfatide-loaded CD1d tetramer to detect type Ⅱ NKT cells in miceChinese Full Text
ZHANG Gu-qin1,NIE Han-xiang2,YANG Jiong1,YU Hong-ying2 1Department of Respiratory Diseases,Zhongnan Hospital,Wuhan University,Wuhan 430071;2Department of Respiratory Diseases,People’ s Hospital of Wuhan University,Wuhan 430060,China
Abstract: AIM: To create a method of detecting typeⅡ natural killer T（NKT） cells of mice.METHODS: Biotinylated mouse CD1d monomers were mixed with sulfatide at a molar ratio of 1∶ 3（protein∶ lipid） and incubated at room temperature overnight,and then 80 μg of streptavidin-PE was added into 200 μg of the CD1d-sulfatide mixture and incubated at room temperature for 4 h to get sulfatide/CD1d tetramer.Flow cytometry was used to detect the percentage of type Ⅱ NKT cells in mononuclear cells（MNCs） of lung and spleen of normal mice,as well as the percentage of type Ⅱ NKT cells in spleen MNCs of mice after stimulated with sulfatide.RESULTS: In normal mice,the percentage of typeⅡ NKT cells accounted for（0.875±0.096）% and（1.175±0.263）% in MNCs of spleen and lung;the percentage in spleen MNCs after activated with sulfatide was（2.75±0.603）%,which significantly increased as compared with that in normal mice（P＜0.01）.CONCLUSION: Sulfatide-loaded CD1d tetramer is an effective method of detecting typeⅡ NKT cells in mice.
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- DOI：
10.13423/j.cnki.cjcmi.006486
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- Classification Code：
R392-33
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