Molecular Urology Laboratory, Division of Urology, and the Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.
Proteomics Clin Appl. 2007 May;1(5):476-85. doi: 10.1002/prca.200700012. Epub 2007 Apr 19.
Cancer sera contain antibodies that react with autologous cellular antigens. Here, we report the use of a novel native antigen-based platform, the "reverse capture" autoantibody microarray, for identification of autoantigens against which autoantibody expression may be used to differentiate between patients with prostate cancer and benign prostate hyperplasia (BPH). Serum samples were collected at our institution from patients with BPH and patients with prostate carcinoma with similar blood prostate-specific antigen levels. IgG was purified from individual prostate cancer sera, differentially labeled with fluorescent dyes, and competitively hybridized against purified IgG from a group of well-characterized BPH control patients on our reverse capture microarray platform. For each experiment, we performed a two-slide dye-swap. Using this platform, we identified 28 unique antigen-autoantibody reactivities that have the potential to discriminate prostate cancer from BPH. These autoantigens, with p-values ≤0.01, can be placed into the following general categories: protein kinases, cell-cycle regulators, cancer-growth factors, apoptosis mediators, and transcription factors. In addition, only 1 of the 28 autoantigens remained differentially targeted by autoantibodies in post-surgical prostate cancer patients after a minimum 1-year follow-up. Autoantibody profiling using this platform may be a useful tool for differentiating between malignant and benign diseases of the prostate.
癌症患者的血清中含有与自身细胞抗原发生反应的抗体。在这里，我们报告了一种新型的基于天然抗原的平台——“反向捕获”自身抗体微阵列，用于鉴定自身抗原，通过检测针对这些自身抗原的自身抗体的表达，可能有助于区分前列腺癌患者和良性前列腺增生（BPH）患者。我们从本机构的 BPH 患者和前列腺癌患者中收集了血清样本，这些患者的血液前列腺特异性抗原水平相似。从个别前列腺癌血清中纯化 IgG，用荧光染料进行差异标记，并在我们的反向捕获微阵列平台上与一组经过充分特征描述的 BPH 对照患者的纯化 IgG 进行竞争性杂交。对于每个实验，我们进行了两次双色荧光染料交换实验。使用该平台，我们鉴定出了 28 种独特的抗原-自身抗体反应，这些反应有可能将前列腺癌与 BPH 区分开来。这些自身抗原的 p 值≤0.01，可以分为以下几类：蛋白激酶、细胞周期调节剂、癌症生长因子、细胞凋亡介质和转录因子。此外，在经过至少 1 年的随访后，只有 1 种自身抗原在接受手术后的前列腺癌患者中仍然被自身抗体特异性靶向。使用该平台进行自身抗体分析可能是区分前列腺良恶性疾病的有用工具。
