Induction of cytotoxic T lymphocytes specific for minor histocompatility antigen through transferringThell receptor gene

在hla匹配供体的造血SCT中，供体和受体之间微小组织相容性抗原（mHas）的差异诱导了GVHD和GVL效应等同种异体免疫反应。根据对同种异体SCT结果的分析，CD62L已被报道为mHa。为了鉴定可能参与GVL效应的CD62L衍生肽，我们制备了所有可能的18种不同的CD62L衍生的9-mer肽，其中9个位置中有一个包含密码子213，并在CD62L不同的SCT患者中测试了这些肽特异性的细胞毒性t细胞前体。将5例移植后发生GVHD的I-ILA-A*2402患者的PBMC与自体单核细胞来源的dc或HLA-A*2402转染的T2细胞孵育，用9-mer肽脉冲，或仅用9-mer肽孵育，孵育16小时后检测PBMC分泌的1fn - γ。每个患者的两到四种不同的受体类型肽诱导的ifn - γ分泌比其他患者高。用肽脉冲的T2-A24细胞刺激PBMC，获得4种肽特异性的t细胞系。培养的T细胞有效地裂解肽脉冲的T2-A24细胞以及HLA-A*2402阳性的与CD62L不同的淋巴母细胞系细胞。加入抗hla I类的单克隆抗体阻断了这种细胞毒性。WIC稳定实验表明，这4种肽结合到细胞表面和HLA复合物稳定。这些结果表明，这些肽可能是能够诱导GVL作用的mHas，并且识别cd62l衍生多态性肽的ctl可能在治疗SCT后复发性白血病方面具有治疗价值。

In hematopoietic SCT from HLA-matched donors, allogeneic immune reactions such as GVHD and GVL effect are induced by disparities in minor histocompatibility antigens (mHas) between donor and recipient. CD62L has been reported to act as an mHa based on analysis of the outcome of allogeneic SCT. To identify peptides derived from CD62L that may be involved in GVL effect, we prepared all possible 18 different 9-mer peptides derived from CD62L that include codon 213 in one of the 9 positions, and tested for cytotoxic T-cell precursors specific to any of these peptides in patients after CD62L-disparate SCT. PBMC obtained from 5 patients with I-ILA-A*2402 who developed GVHD following transplantation were incubated with autologous monocyte-derived DCs or HLA-A*2402-transfected T2 cells which were pulsed with the 9-mer peptides, or incubated with the peptides only, and 1FN-gamma secretion from PBMC after 16 hours incubation was examined. Two to 4 different recipient-type peptides for each patient induced higher IFN-gamma secretion than the others. By stimulating PBMC with the peptide-pulsed T2-A24 cells, T-cell lines specific for 4 peptides were obtained. The cultured T cells efficiently lysed peptide-pulsed T2-A24 cells as well as HLA-A*2402-positive lymphoblastic cell line cells that were disparate with CD62L. Addition of mAbs against HLA-class I blocked this cytotoxicity. WIC stabilization assay demonstrated that the 4 peptides bound to the cell surface and HLA complex stabilized. These results suggest these peptides may serve as mHas capable of inducing GVL effect, and CTLs recognizing CD62L-derived polymorphic peptides may have therapeutic value in treating relapsed leukemia after SCT.