Fernandes D M, Baird A M, Berg L J, Rock K L
Department of Pathology, University of Massachusetts Medical Center, Worcester, MA 01655, USA.
J Immunol. 1999 Aug 1;163(3):1306-14.
E710.2.3 is a murine thymic lymphoma cell line with an immature phenotype (CD4-CD8-) that proliferates in response to thymocytes or PMA when cultured at low density and proliferates spontaneously when grown at high density. To identify functional molecules on this cell line, we screened for mAbs that could block its proliferation. A hamster mAb, DMF10.62.3, inhibited the spontaneous, thymocyte-induced, and PMA-stimulated proliferation of E710.2.3 in vitro and induced these cells to undergo apoptosis. The mAb also caused homotypic aggregation of E710.2.3, which was inhibited by cytochalasin B, trifluoperazine, a combination of sodium azide and 2-deoxyglucose, EDTA, incubation at 4 degrees C, or treatment with paraformaldehyde. The DMF10 62.3 mAb stained a number of immortalized murine and human cell lines and, where tested, blocked their proliferation and caused death to varying extents by apoptosis. The molecule recognized by the mAb DMF10.62.3 was expressed on day 14 fetal thymus Thy1.2-positive cells. However, it was not detected on adult murine thymocytes, splenocytes, or bone marrow cells or on splenic LPS-activated B cells or Con A-activated T cells. The Ab immunoprecipitated a 40-kDa molecule from E710.2.3 that was not glycosylphosphatidylinositol linked. The data suggest that the molecule recognized by DMF62.3 is a novel cell surface molecule that may be involved in cell proliferation and/or cell death.
E710.2.3是一种具有未成熟表型(CD4-CD8-)的小鼠胸腺淋巴瘤细胞系,当在低密度培养时,它会对胸腺细胞或佛波酯(PMA)作出反应而增殖,而在高密度生长时则会自发增殖。为了鉴定该细胞系上的功能分子,我们筛选了能够阻断其增殖的单克隆抗体。一种仓鼠单克隆抗体DMF10.62.3在体外抑制了E710.2.3的自发增殖、胸腺细胞诱导的增殖以及PMA刺激的增殖,并诱导这些细胞发生凋亡。该单克隆抗体还导致E710.2.3发生同型聚集,而细胞松弛素B、三氟拉嗪、叠氮化钠和2-脱氧葡萄糖的组合、乙二胺四乙酸(EDTA)、4℃孵育或多聚甲醛处理均可抑制这种聚集。DMF10 62.3单克隆抗体可对多种永生化的小鼠和人类细胞系进行染色,并且在测试中,它在不同程度上阻断了这些细胞系的增殖并通过凋亡导致细胞死亡。单克隆抗体DMF10.62.3识别的分子在胎龄14天的胸腺Thy1.2阳性细胞上表达。然而,在成年小鼠胸腺细胞、脾细胞或骨髓细胞上,以及在脾脏脂多糖激活的B细胞或刀豆蛋白A激活的T细胞上均未检测到该分子。该抗体从E710.2.3免疫沉淀出一个40 kDa的分子,该分子不是糖基磷脂酰肌醇连接的。数据表明,DMF62.3识别的分子是一种新型细胞表面分子,可能参与细胞增殖和/或细胞死亡。