中文摘要：

目的研究体外扩增小鼠CD8^＋NK1.1^＋NKT细胞的表面分子标志、分化途径及细胞属性。方法获取超抗原SEB活化后体外扩增的小鼠效应细胞,用抗CD3-PerCP、CD4-FITC、CD8-PE、NK1.1-APC、TcRVβ8-FITC和CD69-FITC荧光抗体染色后,用流式细胞仪（FCMCalibueBD,USA）鉴定CD8^＋NK1.1^＋NKT细胞的表面分子标志和分化途径。用逆转录聚合酶链反应测定细胞内各种细胞因子和Foxp3基因转录水平。结果体外扩增的细胞中91.92%是CD8^＋T细胞,其中22.75%的细胞是CD8^＋NN1.1＋NKT细胞,高于正常值（0.21±0.19,n=12）108倍以上。19.61%NKT细胞是TcRVβ8＋NN1.1＋NKT细胞。CD69分子的表达由原始0.11%增加到85.95%。CD4＋T、CD3＋T细胞亚群和CD4＋NKT、CD3＋NKT及CD4-CD8-NKT细胞亚群没有增加。扩增细胞TGF-β的mRNA表达呈阳性,不表达Foxp3和细胞因子IL-2、4、5、6、10及IFN-γ。CD8^＋NKT细胞直接由CD8^＋T细胞分化而来。结论 CD8^＋NK1.1^＋NKT细胞的分子特征为CD69＋Foxp3-TcRVβ8＋TGF-β＋CD8^＋NK1.1^＋;它们既不是CD8^＋T调节细胞（Treg.）,也不是CD4^＋NKT细胞,直接由CD8^＋T细胞分化而来,带有TCRVβ8受体,应属于T细胞亚群。

英文摘要：

Objective To study the surface markers, differentiation pathways and other properties of mice CD8^＋NK1.1^＋NKT cells amplified in vitro. Methods In vitro amplified mice CD8^＋NK1.1^＋NKT cells were obtained by treating mice spleen cells with Staphylococcus enterotoxin B （SEB）. After the cells were stained with fluorescent antibodies, their surface markers and differential pathways were detected with a flow cytometer（Calibue BD, USA）. Gene transcription levels in cytokines and Foxp3 of the cells were measured by RT-PCR. Results Of the in vitro –amplified cells, CD8^＋T cells accounted for 91.92% with 22.75% being CD8^＋NK1.1^＋ NKT cells, which was 108 times higher than that in normal lymphocytes, and NKT cells accounted for 19.61% of TcRVβ8＋ NN1.1＋NKT cells. The expression rate of CD69 was increased from 0.11% to 85.95%. The expression of CD4^＋T, CD8^＋T, CD3＋NKT, CD4^＋NKT and CD4-CD8-NKT cell subsets did not increase. The mRNA was positively expressed in amplified TGF-βcells which did not express Foxp3 and cytokines including IL-2, 4-6, 10 and IFN-γ. CD8^＋NKT cells were directly differentiated from CD8^＋T cells. Conclusion CD8^＋NK1.1^＋NKT cells, characterized by CD69＋Foxp3-TcRVβ8＋ TGF-β＋CD8^＋NK1.1^＋, are neither CD8^＋ regulatory cells（Treg,） nor CD4^＋NKT cells. They are directly differentiated from CD8^＋T cells and carry TCRVβ8 receptors, thus belonging to a subset of T lymphocytes.