McKenzie J L, Fabre J W
J Immunol. 1981 Mar;126(3):843-50.
BALB/c mice were immunized with purified human brain Thy-1 and used in cell fusion experiments to derive an anti-human Thy-1 monoclonal antibody. Specificity was proven by a) showing that the antigen recognized had precisely the tissue distribution expected of human Thy-1 from previous studies, and b) demonstrating that the molecule purified from human brain by monoclonal antibody affinity columns 1) had the same mobility on SDS PAGE as pure rat Thy-1 and 2) could inhibit an assay previously shown to be directed at the human-rat cross-reactive component of Thy-1. Studies with the fluorescence-activated cell sorter confirmed that Thy-1 is absent from human blood lymphocytes and cell suspensions from human lymphoid organs, except that very weak fluorescence could be detected on 7% of thymus cells in suspension. However, fluorescence studies on frozen sections showed bright staining restricted mainly to the periphery of the thymus lobule, the marginal zone and some periarteriolar lymphocytes in the spleen, and the post-capillary venules of lymph node. In some lymph nodes, a halo of Thy-1 positive cells was seen around lymphatic nodules or germinal centers, but this was not a constant finding. This distribution is different from the known distribution of Thy-1 in the lymphoid tissues of the mouse and rat. Our studies suggest that human Thy-1 might be involved in lymphocyte recirculation, and that it is a marker for early T lymphocytes in man. The studies also show that the few Thy-1 positive cells in human lymphoid organs are selectively lost in the preparation of single-cell suspensions.
用纯化的人脑 Thy-1 免疫 BALB/c 小鼠,并将其用于细胞融合实验以获得抗人 Thy-1 单克隆抗体。通过以下方式证明其特异性:a) 表明所识别的抗原具有先前研究中预期的人 Thy-1 精确组织分布;b) 证明通过单克隆抗体亲和柱从人脑中纯化的分子 1) 在 SDS-PAGE 上与纯大鼠 Thy-1 具有相同的迁移率,并且 2) 能够抑制先前显示针对 Thy-1 的人-大鼠交叉反应成分的检测。荧光激活细胞分选仪研究证实,人血淋巴细胞和人淋巴器官的细胞悬液中不存在 Thy-1,不过在悬浮的 7% 的胸腺细胞上可检测到非常微弱的荧光。然而,对冰冻切片的荧光研究显示,明亮染色主要局限于胸腺小叶周边、脾脏的边缘区和一些动脉周围淋巴细胞以及淋巴结的毛细血管后微静脉。在一些淋巴结中,在淋巴小结或生发中心周围可见 Thy-1 阳性细胞晕,但这并非恒定现象。这种分布与已知的 Thy-1 在小鼠和大鼠淋巴组织中的分布不同。我们的研究表明,人 Thy-1 可能参与淋巴细胞再循环,并且它是人类早期 T 淋巴细胞的标志物。研究还表明,人淋巴器官中少数 Thy-1 阳性细胞在单细胞悬液制备过程中会选择性丢失。