WT1特异性 CD8+ T 细胞治疗乳腺癌的实验研究

摘要： 目的 通过检测 Wilms’肿瘤基因 1 （WT1） 特异性 CD8+ T 细胞对乳腺癌细胞的杀伤活性， 探讨正常外周血中提取的 WT1 特异性 CD8+ T 细胞用于治疗乳腺癌的可行性。方法 运用流式细胞仪检测 20 例人白细胞抗原（HLA） -A2 血清阳性健康志愿者外周血中 WT1 特异性 CD8+ T 细胞的浓度， 通过 WT1/主要组织相容性复合体（MHC） 连锁状多聚体磁珠分选 WT1 特异性 CD8+ T 细胞并进行培养， 细胞毒性实验检测 WT1 特异性 CD8+ T 细胞的功能。结果 20 例 HLA-A2 血清阳性健康志愿者外周血中均可检测到 WT1 特异性 CD8+ T 细胞， 12 例健康外周血中 WT1 特异性 CD8+ T 细胞的浓度＞0.5%， 其中 4 例＞1%。WT1/MHC 连锁状多聚体磁珠分选后， 其浓度明显增加至 80%左右。WT1 特异性 CD8+ T 细胞可特异性杀伤 WT1 多肽负载的乳腺癌细胞株。结论 健康志愿者外周血中可检测到 WT1 特异性 CD8+ T 细胞， 该细胞可能对乳腺癌细胞具有特异性杀伤作用， 提示 WT1 特异性 CD8+ T 细胞用于乳腺癌过继性免疫治疗的可能性。

Experimental study of WT1 specific CD8+ T cells in the treatment of breast cancer

Abstract: Objective To investigate the feasibility of Wilms’tumor gene 1 (WT1)- specific CD8+ T cells from periph⁃ eral blood for the treatment of breast cancer by detecting the killing activity of WT1 specific CD8+ T cells on breast cancer cells. Methods Flow cytometry was used to detect WT1-specific CD8+ T cells in the peripheral blood of 20 samples from HLA-A2 seropositive healthy donors, which were isolated by WT1/MHC streptamer magnetic beads and cultured. The func⁃ tion of WT1-specific CD8+ T cells were analysis by cytotoxicity assay. Results Twelve of 20 healthy donors had naive WT1-specific CD8+ T-cell frequencies of ＞0.5%, and 4 of 20 even ＞1.0% of all CD8+ T cells. After positive selection by magnetic cell separation, a purity of up to 80% can be achieved. WT1 specific CD8+ T cells can specifically kill breast can⁃ cer cell line with WT1 polypeptide. Conclusion WT1 specific CD8+ T cells can be detected in peripheral blood of healthy volunteers. WT1 specific CD8+ T cells have killing effect on breast cancer cells, suggesting the feasibility of adoptive immu⁃ notherapy for breast cancer.