Garrido M C, Cordell J L, Becker M H, Key G, Gerdes J, Jones M, Gatter K C, Mason D Y
Nuffield Department of Pathology, John Radcliffe Hospital, Oxford.
J Clin Pathol. 1992 Oct;45(10):860-5. doi: 10.1136/jcp.45.10.860.
To characterise a newly developed mouse monoclonal antibody JC1 which recognises a nuclear antigen present in proliferating cells in normal tissues and neoplastic lesions, and which is absent in resting cells.
The methodology was established using a representative range of frozen sections from normal tissues and from certain tumours which were immunostained with antibodies Ki67 and JC1. The molecular weight of the antigen recognised by JC1 was obtained by western blot analysis and this was compared with that of Ki67. IM-9 cell lysates containing Ki67 derived plasmids were also tested with JC1 antibody.
Biochemical investigation indicated that the antigen recognised by JC1 gives two molecular weight bands of 212 and 123 kilodaltons, which is distinct from the well characterised anti-proliferation monoclonal antibody Ki67 (395-345 kilodaltons). In addition recombinant Ki67 protein is not recognised by JC1. Immunohistological reactivity was seen in areas known to contain numerous proliferating cells such as lymphoid germinal centres, splenic white matter, cortical thymocytes and undifferentiated spermatogonia. In tumours many cells from adenocarcinomas, oat cell carcinomas, squamous cell carcinomas of lung, and seminomas were labelled by JC1 with a distribution and proportion similar to that seen with Ki67. In normal tissues the only apparent difference was in testis where JC1 stained a considerably greater number of cells than Ki67. In all cases studied the new antibody showed nuclear reactivity only. JC1 did not show any cytoplasmic crossreactivity with squamous cells as is frequently seen with Ki67.
Antibody JC1, which recognises a nuclear antigen present in proliferating cells, should provide a useful adjunct to Ki67 as a marker of proliferation especially in those cases such as squamous cell carcinomas where a Ki67 index cannot be determined.
鉴定一种新开发的小鼠单克隆抗体JC1，该抗体可识别正常组织和肿瘤性病变中增殖细胞内存在的一种核抗原，而静止细胞中不存在该抗原。
使用来自正常组织和某些肿瘤的一系列代表性冰冻切片建立方法，并用抗体Ki67和JC1进行免疫染色。通过蛋白质印迹分析获得JC1识别的抗原的分子量，并与Ki67的分子量进行比较。还使用JC1抗体检测了含有Ki67衍生质粒的IM-9细胞裂解物。
生化研究表明，JC1识别的抗原产生两条分子量分别为212和123千道尔顿的条带，这与特征明确的抗增殖单克隆抗体Ki67（395-345千道尔顿）不同。此外，重组Ki67蛋白不被JC1识别。在已知含有大量增殖细胞的区域，如淋巴生发中心、脾白质、皮质胸腺细胞和未分化精原细胞中观察到免疫组织学反应性。在肿瘤中，许多来自腺癌、燕麦细胞癌、肺鳞状细胞癌和精原细胞瘤的细胞被JC1标记，其分布和比例与Ki67所见相似。在正常组织中，唯一明显的差异在于睾丸，JC1染色的细胞数量比Ki67多得多。在所研究的所有病例中，新抗体仅显示核反应性。JC1与鳞状细胞没有任何细胞质交叉反应，而Ki67经常出现这种情况。
识别增殖细胞中存在的核抗原的抗体JC1，应作为Ki67的有用辅助物，作为增殖的标志物，尤其是在无法确定Ki67指数的鳞状细胞癌等病例中。
