Department of Hematology, Leiden University Medical Center, Leiden, the Netherlands.
Cancer Res. 2010 Nov 15;70(22):9073-83. doi: 10.1158/0008-5472.CAN-10-1832. Epub 2010 Nov 9.
Patients with malignant diseases can be effectively treated with allogeneic hematopoietic stem cell transplantation (allo-SCT). Polymorphic peptides presented in HLA molecules, the so-called minor histocompatibility antigens (MiHA), play a crucial role in antitumor immunity as targets for alloreactive donor T cells. Identification of multiple MiHAs is essential to understand and manipulate the development of clinical responses after allo-SCT. In this study, CD8+ T-cell clones were isolated from leukemia patients who entered complete remission after allo-SCT, and MiHA-specific T-cell clones were efficiently selected for analysis of recognition of a panel of EBV-transformed B cells positive for the HLA restriction elements of the selected T-cell clones. One million single nucleotide polymorphisms (SNP) were determined in the panel cell lines and investigated for matching with the T-cell recognition data by whole genome association scanning (WGAs). Significant association with 12 genomic regions was found, and detailed analysis of genes located within these genomic regions revealed SNP disparities encoding polymorphic peptides in 10 cases. Differential recognition of patient-type, but not donor-type, peptides validated the identification of these MiHAs. Using tetramers, distinct populations of MiHA-specific CD8+ T cells were detected, demonstrating that our WGAs strategy allows high-throughput discovery of relevant targets in antitumor immunity after allo-SCT.
患有恶性疾病的患者可以通过异体造血干细胞移植(allo-SCT)进行有效治疗。在 HLA 分子中呈递的多态性肽,即所谓的次要组织相容性抗原(MiHA),作为同种异体反应性供体 T 细胞的靶标,在抗肿瘤免疫中发挥着关键作用。鉴定多种 MiHA 对于理解和操纵 allo-SCT 后临床反应的发展至关重要。在这项研究中,从接受 allo-SCT 后进入完全缓解的白血病患者中分离出 CD8+T 细胞克隆,并有效地选择 MiHA 特异性 T 细胞克隆,以分析对一组 EBV 转化的 B 细胞的识别,这些 B 细胞阳性表达所选 T 细胞克隆的 HLA 限制元件。在面板细胞系中确定了 100 万个单核苷酸多态性(SNP),并通过全基因组关联扫描(WGAs)调查它们与 T 细胞识别数据的匹配情况。发现与 12 个基因组区域存在显著关联,对这些基因组区域内基因的详细分析显示,在 10 个病例中存在编码多态性肽的 SNP 差异。对患者型而不是供体型肽的差异识别验证了这些 MiHA 的鉴定。使用四聚体,检测到 MiHA 特异性 CD8+T 细胞的独特群体,证明我们的 WGAs 策略允许在 allo-SCT 后抗肿瘤免疫中高通量发现相关靶标。