Department of Gastroenterological Surgery II, Hokkaido University Graduate School of Medicine, and Department of Thoracic Surgery, Sapporo-Minamisanjo Hospital, Sapporo, Hokkaido 060-8638, Japan.
Oncol Rep. 2013 Apr;29(4):1315-21. doi: 10.3892/or.2013.2275. Epub 2013 Feb 5.
This report proposes a concept for the standardization of immunohistochemical evaluation. Immunohistochemical staining has several problems associated with the sensitivity of the technical process and standardization of the assessment of potent staining. We provided data focusing on this concept through immunostaining for CD154 in non-small cell lung cancer (NSCLC). We used two types of anti-CD154 antibody as primary antibodies in immunohistochemical staining, as previously reported. Western blot analysis confirmed strong CD154 expression in the cultured cell line PC10, but not in LK2. We also assessed CD154 expression in SCID mouse xenografts of these cell lines. SCID xenograft data on western blot analysis were consistent with those of cultured cell lines. These xenografts could thus be used as positive or negative tissue controls for CD154 immunostaining. Primary antibodies should therefore be confirmed as recognizing target lesions, while control tissue specimens should be objectively confirmed as having target products using another experimental method. Our method would allow results to be unified at more than one laboratory and could act as an objective control assessment method in immunohistochemistry.
本报告提出了免疫组织化学评估标准化的概念。免疫组织化学染色存在一些问题，与技术过程的敏感性和有效染色评估的标准化有关。我们通过非小细胞肺癌（NSCLC）中 CD154 的免疫染色提供了关注这一概念的数据。我们使用了两种类型的抗 CD154 抗体作为免疫组织化学染色的一抗，如前所述。Western blot 分析证实，在培养的细胞系 PC10 中 CD154 表达较强，但在 LK2 中没有表达。我们还评估了这些细胞系在 SCID 小鼠异种移植中的 CD154 表达。SCID 异种移植的 Western blot 分析数据与培养细胞系一致。因此，这些异种移植物可作为 CD154 免疫染色的阳性或阴性组织对照。因此，一抗应确认为识别靶病变，而对照组织标本应使用另一种实验方法客观确认为具有靶产物。我们的方法将允许在多个实验室统一结果，并可作为免疫组织化学中的客观控制评估方法。
