Department of Ophthalmology, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, PR China.
Mol Med Rep. 2011 Jan-Feb;4(1):163-7. doi: 10.3892/mmr.2010.397. Epub 2010 Nov 17.
B7-H1, a recently described B7 family member, has been reported to negatively regulate T-cell function in most cancer cells. In this study, we sought to investigate B7-H1 expression in four uveal melanoma (UM) cells (OCM1, SP6.5, OM431 and VUP) to determine the functional significance of B7-H1 expression in T-cell immune response. Using flow cytometry (FCM), we demonstrated that SP6.5 cells had high B7-H1 protein expression, while the other three UM cell lines had none. However, all four UM cell lines expressed B7-H1 mRNA, as confirmed by reverse transcription-polymerase chain reaction. In co-culture experiments using B7-H1-expressing UM cells with T-cells, FCM to determine CD69 expression in T-cells revealed that SP6.5 cell-related B7-H1 inhibited T-cell activation. This effect was eliminated by B7-H1-targeted RNA interference. An Annexin V/PI double staining assay further showed that B7-H1 expressed by SP6.5 cells did not increase the apoptosis of T-cells, though it was found in a variety of other solid tumors. In conclusion, all the UM cell lines constitutively expressed B7-H1 mRNA, while B7-H1 protein was expressed at different levels. UM-related B7-H1 expression negatively regulated T-cell immune response through the inhibition of T-cell activation, and not through the promotion of T-cell apoptosis. This provides new insight into anti-tumor immunity against B7-H1-expressing UM cells.
B7-H1 是一种新发现的 B7 家族成员,据报道在大多数癌细胞中可负向调控 T 细胞功能。在本研究中,我们试图检测 4 株葡萄膜黑色素瘤(UM)细胞(OCM1、SP6.5、OM431 和 VUP)中 B7-H1 的表达,以明确 B7-H1 表达对 T 细胞免疫应答的功能意义。采用流式细胞术(FCM),我们证实 SP6.5 细胞具有高水平的 B7-H1 蛋白表达,而其它 3 株 UM 细胞系则无表达。然而,通过逆转录-聚合酶链反应(RT-PCR)证实 4 株 UM 细胞系均表达 B7-H1 mRNA。在与表达 B7-H1 的 UM 细胞共培养的 T 细胞中,通过 FCM 检测 T 细胞 CD69 的表达,我们发现 SP6.5 细胞相关的 B7-H1 抑制 T 细胞的激活。该作用可被 B7-H1 靶向的 RNA 干扰消除。Annexin V/PI 双染实验进一步显示,SP6.5 细胞表达的 B7-H1 虽未增加 T 细胞的凋亡(此现象在多种其它实体瘤中存在),但可抑制 T 细胞的激活。综上所述,所有 UM 细胞系均持续表达 B7-H1 mRNA,但其 B7-H1 蛋白的表达水平存在差异。UM 相关的 B7-H1 表达通过抑制 T 细胞的激活,而非促进 T 细胞的凋亡,负向调控 T 细胞免疫应答。这为针对表达 B7-H1 的 UM 细胞的抗肿瘤免疫提供了新的见解。