Peled A, Shezen E, Schwartz D, Shav-Tal Y, Kushtai G, Lee B C, Gothelf Y, Krupsky M, Zipori D
Department of Neurology, Hadassah-Hebrew University Hospital, Jerusalem, Israel.
Hybridoma. 1997 Aug;16(4):325-34. doi: 10.1089/hyb.1997.16.325.
We describe a novel mouse monoclonal antibody (PRA-72) that recognizes a nuclear antigen associated with cell proliferation. The monoclonal antibody stained the nuclei of logarithmically growing cultured stromal cells. The nuclear staining disappeared when these cells entered Gzero phase of the cell cycle. Western blot analysis revealed a nuclear protein which appeared as a doublet at 35-40 KD, which was undetectable in extracts from confluent cells. Immunocytological study of purified cell populations from various cell cycle phases revealed peripheral nuclear staining in all stages except mitosis, when the chromosomes were observed enveloped with the antigen. In co-cultures of quiescent stromal cells and proliferating hemopoietic precursors, only the latter showed nuclear staining by PRA-72 monoclonal antibody. Further indications for selective expression of the antigen by proliferating cells were found by an immunohistochemical study of various tissues including newborn mouse bone marrow and its surrounding connective tissue, mouse tongue epithelium, and human carcinoma of the colon. This antibody may, therefore, prove useful in the evaluation of human tumors.
我们描述了一种新型小鼠单克隆抗体(PRA - 72),它可识别一种与细胞增殖相关的核抗原。该单克隆抗体可对对数生长期培养的基质细胞核进行染色。当这些细胞进入细胞周期的G0期时,核染色消失。蛋白质印迹分析显示一种核蛋白,其在35 - 40 KD处呈现为双峰,在汇合细胞的提取物中无法检测到。对来自不同细胞周期阶段的纯化细胞群体进行免疫细胞学研究发现,除有丝分裂阶段(此时观察到染色体被该抗原包裹)外,在所有阶段均可见核周染色。在静止基质细胞与增殖造血前体细胞的共培养中,只有后者显示出被PRA - 72单克隆抗体进行核染色。通过对包括新生小鼠骨髓及其周围结缔组织、小鼠舌上皮和人类结肠癌在内的各种组织进行免疫组织化学研究,发现了增殖细胞选择性表达该抗原的进一步证据。因此,这种抗体可能在人类肿瘤评估中证明有用。