Enzymatic discovery of a HER-2/neu epitope that generates cross-reactive T cells.
Enzymatic discovery of a HER-2/neu epitope that generates cross-reactive T cells.
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DOI：
10.4049/jimmunol.1201264
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发表时间：
2013-01-01
期刊：
影响因子：
--
通讯作者：
Knutson KL
中科院分区：
文献类型：
--
作者：
Henle AM;Erskine CL;Benson LM;Clynes R;Knutson KL
表达 HER-2/neu 的乳腺癌患者在标准治疗后仍面临复发风险。针对 HER-2/neu 预防复发的疫苗正处于临床测试的各个阶段。许多疫苗都含有 HER-2/neu HLA-A2 结合肽 p369-377 (KIFGSLAFL)，因为研究表明，对该表位具有特异性的细胞毒性 T 淋巴细胞 (CTL) 可以直接杀死 HER-2/neu 过度表达的乳腺癌细胞。因此，了解肿瘤如何处理该表位对于识别将从免疫接种中受益的患者可能很重要。蛋白酶体制剂用于确定 p369-377 是否由较大的 HER-2/neu 衍生片段加工而成。使用 HPLC、质谱、细胞毒性测定、IFN-γ ELIspot 和人乳腺癌细胞系来评估蛋白水解片段。纯化的 20S 蛋白酶体和免疫蛋白酶体未检测到 p369-377 的加工，这表明肿瘤细胞可能无法加工 HER-2/neu 蛋白中的该抗原并将其呈递在 HLA I 类环境中。相反，我们表明其他胞外结构域 HER-2/neu 肽序列始终由蛋白酶体加工。其中一个序列 p373-382 (SLAFLPESFD) 与 HLA-A2 的结合强于 p369-377。 p373-382 特异的 CTL 可识别与 HLA-A2 复合的 p373-382 和 p369-377。 p373-382 特异性 CTL 也比 p369-377 特异性 CTL 更高水平地杀死人乳腺癌细胞系。相反，p369-377 特异的 CTL 识别 p373-382。肽 p373-382 是乳腺癌疫苗的候选表位，因为它由蛋白酶体加工并结合 HLA-A2。
Patients with HER-2/neu-expressing breast cancer remain at risk for relapse following standard therapy. Vaccines targeting HER-2/neu to prevent relapse are in various phases of clinical testing. Many vaccines incorporate the HER-2/neu HLA-A2 binding peptide p369–377 (KIFGSLAFL), since it has been shown that cytotoxic T lymphocytes (CTLs) specific for this epitope can directly kill HER-2/neu overexpressing breast cancer cells. Thus, understanding how tumors process this epitope may be important for identifying the patients that would benefit from immunization. Proteasome preparations were used to determine if p369–377 was processed from larger HER-2/neu derived fragments. HPLC, mass spectrometry, cytotoxicity assays, IFN-γ ELIspot, and human breast cancer cell lines were used to assess the proteolytic fragments. Processing of p369–377 was not detected by purified 20S proteasome and immunoproteasome, indicating that tumor cells may not be capable of processing this antigen from the HER-2/neu protein and presenting it in the context of HLA class I. Instead, we show that other extracellular domain HER-2/neu peptide sequences are consistently processed by the proteasomes. One of these sequences, p373–382 (SLAFLPESFD), bound HLA-A2 stronger than p369–377. CTLs specific for p373–382 recognized both p373–382 and p369–377 complexed with HLA-A2. CTL specific for p373–382 also killed human breast cancer cell lines at higher levels than p369–377 specific CTL. Conversely, CTLs specific for p369–377 recognized p373–382. Peptide p373–382 is a candidate epitope for breast cancer vaccines as it is processed by proteasomes and binds HLA-A2.
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