摘要
目的进一步认识Vα24 NKT细胞在抗肿瘤免疫中的作用。方法从正常人外周血中扩增并纯化得到Vα24 NKT细胞,通过流式细胞术检测其表型、活化后的细胞因子、细胞坏死相关因子的表达情况,采用DIOC18染色及流式细胞术测定Vα24 NKT细胞对肿瘤细胞的杀伤率,并利用不同的阻断剂进行阻断实验。结果Vα24 NKT细胞分泌高水平的IFN-γ和IL-4,并表达TNF-α、穿孔素(perforin)、FasL、TRAIL等细胞坏死相关因子。Vα24 NKT细胞对表面表达CD1d分子的肿瘤细胞株的杀伤率高于不表达CD1d分子的肿瘤细胞株,同时在α-GalCer存在时,Vα24 NKT细胞的杀伤活性增强。对Perforin、FasL、TNF-α、TRAIL进行阻断后,ConA对Vα24 NKT细胞的杀伤作用的阻断效果最为明显。结论Vα24 NKT对肿瘤的杀伤作用是CD1d分子限制性和抗原特异性的。Vα24 NKT通过穿孔素途径和TNF家族成员发挥其细胞毒作用,而穿孔素途径是其肿瘤杀伤的最主要途径。
Objective：To further recogrige the role of the Vα24 NKT cells in anti-tumor immunity. Methods： The Vα24 NKT cells were proliferated and purified from normal human peripheral blood. The phenotypes, the expressions of both activated cytokines and necrosis-related factors on Vα24 NKT cells were determined by flow cytometry. The cytotoxicity of Vα24NKT cells to tumor cells was measured by DIOC18 staining and flow cytometry. Different inhibitors were used in the blocking experiments. Results： Vα24 NKT cells secreted high levels of interferon-γ（IFN-γ） and interleukin-4 （IL-4） and expressed necrosis-related factors such as TNFα, perforin, FasL, and TRAIL. In addition, Vα24NKT cells had increased cytotoxic effect on CD1d-positive tumor cells than CD1 d-negative tumor cells. And the killing activity was enhanced in the presence of αGalCer. When Perforin, FasL, TNFα, and TRAIL were blocked, the blocking effect of ConA against the cytotoxic effects of Vα24 NKT cells were most apparent. Conclusion ： The tumor-killing activity of Vα24NKT cells was CD1 d-dependent and antigen-specific. The cytotoxic effect of Vα24NKT cells is achieved via perforin and TNF family members pathway. However, perforin is the main pathway for its anti-tumor effect.
出处
《肿瘤》
CAS
CSCD
北大核心
2007年第2期83-87,共5页
Tumor
基金
上海市自然科学基金资助项目(01ZB14057)
