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表达普通急性淋巴细胞白血病抗原(CALLA)的胎儿造血细胞的纯化与特性分析

Purification and characterization of fetal hematopoietic cells that express the common acute lymphoblastic leukemia antigen (CALLA).

作者信息

Hokland P, Rosenthal P, Griffin J D, Nadler L M, Daley J, Hokland M, Schlossman S F, Ritz J

出版信息

J Exp Med. 1983 Jan 1;157(1):114-29. doi: 10.1084/jem.157.1.114.

Abstract

Fetal hematopoietic cells that express the common acute lymphoblastic leukemia antigen (CALLA) were purified from both fetal liver and fetal bone marrow by immune rosetting with sheep erythrocytes coated with rabbit anti-mouse immunoglobulin and by fluorescence-activated cell sorting. Dual fluorescence techniques disclosed that these cells were heterogenous with respect to the expression of a series of differentiation and activation antigens defined by monoclonal antibodies. Thus, whereas all CALLA+ cells were Ia+ and expressed two activation antigens, J2 and T10, only 30-50% expressed B1 antigen. Furthermore, using methanol-fixed cells, it could be shown that approximately 20% contained intracytoplasmic mu chains (cyto-mu) and that approximately 15% were positive for the terminal transferase enzyme (TdT) marker. The CALLA+ fetal cells thus closely resemble the childhood acute lymphoblastic leukemia cell with respect to surface marker phenotype. A population of CALLA- cells devoid of mature erythroid and myeloid surface markers was found to contain higher numbers of TdT+ cells but lower numbers of cyto-mu, B1, and Ia+ cells than the CALLA+ subset. In vitro analysis of normal, purified CALLA+ cells demonstrated that incubation at 37 degrees C with J5 monoclonal antibody specific for CALLA resulted in the specific modulation of surface antigen. Similar results have previously been obtained with CALLA+ tumor cells. Although phenotypic analysis of CALLA+ cells suggests that these cells are relatively immature lymphoid cells, CALLA+ cells do not appear to contain either myeloid precursor cells (CFU-G/M) or the earliest lymphoid stem cells.

摘要

通过用包被有兔抗小鼠免疫球蛋白的绵羊红细胞进行免疫玫瑰花结试验以及荧光激活细胞分选,从胎儿肝脏和胎儿骨髓中纯化出表达普通急性淋巴细胞白血病抗原(CALLA)的胎儿造血细胞。双荧光技术显示,这些细胞在由单克隆抗体定义的一系列分化和激活抗原的表达方面是异质的。因此,虽然所有CALLA+细胞都是Ia+并表达两种激活抗原J2和T10,但只有30 - 50%表达B1抗原。此外,使用甲醇固定的细胞可以显示,大约20%含有胞质内μ链(胞质μ),大约15%的末端转移酶(TdT)标记呈阳性。因此,CALLA+胎儿细胞在表面标志物表型方面与儿童急性淋巴细胞白血病细胞非常相似。发现一群缺乏成熟红细胞和髓细胞表面标志物的CALLA - 细胞比CALLA+亚群含有更多的TdT+细胞,但胞质μ、B1和Ia+细胞数量较少。对正常纯化的CALLA+细胞进行的体外分析表明,在37℃下用针对CALLA的J5单克隆抗体孵育会导致表面抗原的特异性调节。以前用CALLA+肿瘤细胞也得到了类似的结果。虽然对CALLA+细胞的表型分析表明这些细胞是相对不成熟的淋巴细胞,但CALLA+细胞似乎既不包含髓系前体细胞(CFU - G/M)也不包含最早的淋巴干细胞。

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