Identification of breast cancer peptide epitopes presented by HLA-A*0201
Identification of breast cancer peptide epitopes presented by HLA-A*0201
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DOI：
10.1021/pr700761w
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发表时间：
2008-04-01
影响因子：
4.4
通讯作者：
Hildebrand, William H.
中科院分区：
文献类型：
--
作者：
Hawkins, Oriana E.;VanGundy, Rodney S.;Hildebrand, William H.
细胞免疫机制通过人白细胞抗原（HLA）I类分子检测和破坏癌细胞和感染细胞，所述人白细胞抗原（HLA）I类分子在所有有核细胞的表面上呈递细胞内来源的肽。新的肿瘤特异性表位的鉴定是乳腺癌免疫治疗药物开发的关键步骤。为了直接鉴定癌细胞特有的肽表位，通过缺失HLA A*0201的跨膜和胞质结构域来构建分泌的人I类HLA分子（sHLA）。将得到的sHLA-A*0201转移并在乳腺癌细胞系MCF-7、MDA-MB-231和BT-20以及永生的非致瘤细胞系MCF 10A中表达。将稳定的转染子接种到生物反应器中以产生>25 mg的sHLA-A*0201。通过质谱分析从亲和纯化的sHLA洗脱的肽。HLA-A*0201肽的比较分析揭示了乳腺癌细胞上独特呈现的5个先前未表征的表位。这些肽衍生自在乳腺癌发生和进展中具有明确定义或推定作用的细胞内蛋白：细胞周期蛋白依赖性激酶2（Cdk 2）、鸟氨酸脱羧酶（ODC 1）、动粒相关2（KNTC 2或HEC 1）、巨噬细胞迁移抑制因子（MIF）和外泌体组分6（EXOSC 6）。通过四聚体染色和IFN-γ ELISPOT证明了循环CD 8+细胞对MIF、KNTC 2、EXOSC 6和Cdk 2肽的细胞识别。对I类乳腺癌细胞特有的肽的鉴定和表征为免疫诊断工具和治疗剂的开发提供了推定的靶点。
Cellular immune mechanisms detect and destroy cancerous and infected cells via the human leukocyte antigen (HLA) class I molecules that present peptides of intracellular origin on the surface of all nucleated cells. The identification of novel, tumor-specific epitopes is a critical step in the development of immunotherapeutics for breast cancer. To directly identify peptide epitopes unique to cancerous cells, secreted human class I HLA molecules (sHLA) were constructed by deletion of the transmembrane and cytoplasmic domain of HLA A*0201. The resulting sHLA-A*0201 was transferred and expressed in breast cancer cell lines MCF-7, MDA-MB-231, and BT-20 as well as in the immortal, nontumorigenic cell line MCF10A. Stable transfectants were seeded into bioreactors for production of >25 mg of sHLA-A*0201. Peptides eluted from affinity purified sHLA were analyzed by mass spectroscopy. Comparative analysis of HLA-A*0201 peptides revealed 5 previously uncharacterized epitopes uniquely presented on breast cancer cells. These peptides were derived from intracellular proteins with either well-defined or putative roles in breast cancer development and progression: Cyclin Dependent Kinase 2 (Cdk2), Ornithine Decarboxylase (ODC1), Kinetochore Associated 2 (KNTC2 or HEC1), Macrophage Migration Inhibitory Factor (MIF), and Exosome Component 6 (EXOSC6). Cellular recognition of the MIF, KNTC2, EXOSC6, and Cdk2 peptides by circulating CD8+ cells was demonstrated by tetramer staining and IFN-gamma ELISPOT. The identification and characterization of peptides unique to the class I of breast cancer cells provide putative targets for the development of immune diagnostic tools and therapeutics.