Jaramillo André, Narayanan Kishore, Campbell Lacey G, Benshoff Nicholas D, Lybarger Lonnie, Hansen Ted H, Fleming Timothy P, Dietz Jill R, Mohanakumar T
Department of Surgery, Washington University School of Medicine, St. Louis, MO 63110, USA.
Breast Cancer Res Treat. 2004 Nov;88(1):29-41. doi: 10.1007/s10549-004-8918-1.
A breast cancer-associated antigen, mammaglobin-A, is specifically expressed in 80% of primary breast tumors. The definition of immune responses against this highly expressed breast cancer-specific antigen should be of great value in the development of new therapeutic strategies for breast cancer. Thus, the purpose of this study was to identify HLA-A2-restricted mammaglobin-A-derived epitopes recognized by CD8+ cytotoxic T lymphocytes (CTL). We identified seven mammaglobin-A-derived candidate epitopes that bind the HLA-A2 molecule (Mam-A2.1-7) by means of a HLA class I-peptide binding computer algorithm from the Bioinformatics & Molecular Analysis Section of the National Institutes of Health. Subsequently, we determined that CD8+ CTLs from breast cancer patients reacted to the Mam-A2.1 (83-92, LIYDSSLCDL), Mam-A2.2 (2-10, KLLMVLMLA), Mam-A2.3 (4-12, LMVLMLAAL), Mam-A2.4 (66-74, FLNQTDETL), and Mam-A2.7 (32-40, TINPQVSKT) epitopes using an IFN-gamma ELISPOT assay. Interestingly, healthy individuals also showed high reactivity to the Mam-A2.2 epitope. Two CD8+ CTL lines generated in vitro against TAP-deficient T2 cells loaded with the candidate epitopes showed significant cytotoxic activity against the Mam-A2.1-4 epitopes. These CD8+CTL lines recognized a HLA-A2+breast cancer cell line expressing the Mam-A2.1 epitope. In addition, DNA vaccination of HLA-A2+/human CD8+ double-transgenic mice with a DNA construct encoding the Mam-A2.1 epitope and the HLA-A2 molecule induced a significant expansion of epitope-specific CD8+ CTLs that recognize the same HLA- A2+/Mam-A2.1+ breast cancer cell line. In conclusion, these results demonstrate the immunotherapeutic potential of mammaglobin-A for the treatment and prevention of breast cancer.
一种与乳腺癌相关的抗原，乳腺珠蛋白-A，在80%的原发性乳腺肿瘤中特异性表达。针对这种高表达的乳腺癌特异性抗原的免疫反应的定义，对于开发新的乳腺癌治疗策略具有重要价值。因此，本研究的目的是鉴定被CD8+细胞毒性T淋巴细胞（CTL）识别的HLA-A2限制性乳腺珠蛋白-A衍生表位。我们通过美国国立卫生研究院生物信息学与分子分析部门的HLA I类肽结合计算机算法，鉴定出七个与HLA-A2分子结合的乳腺珠蛋白-A衍生候选表位（Mam-A2.1-7）。随后，我们使用干扰素-γ ELISPOT检测法确定，来自乳腺癌患者的CD8+ CTL对Mam-A2.1（83-92，LIYDSSLCDL）、Mam-A2.2（2-10，KLLMVLMLA）、Mam-A2.3（4-12，LMVLMLAAL）、Mam-A2.4（66-74，FLNQTDETL）和Mam-A2.7（32-40，TINPQVSKT）表位有反应。有趣的是，健康个体对Mam-A2.2表位也表现出高反应性。在体外针对负载候选表位的TAP缺陷型T2细胞产生的两条CD8+ CTL系，对Mam-A2.1-4表位显示出显著的细胞毒性活性。这些CD8+ CTL系识别表达Mam-A2.1表位的HLA-A2+乳腺癌细胞系。此外，用编码Mam-A2.1表位和HLA-A2分子的DNA构建体对HLA-A2+/人CD8+双转基因小鼠进行DNA疫苗接种，诱导了识别相同HLA-A2+/Mam-A2.1+乳腺癌细胞系的表位特异性CD8+ CTL的显著扩增。总之，这些结果证明了乳腺珠蛋白-A在乳腺癌治疗和预防中的免疫治疗潜力。
