Zwerner R K, Barstad P A, Acton R T
J Exp Med. 1977 Oct 1;146(4):986-1000. doi: 10.1084/jem.146.4.986.
The Thy-l.1 molecule was isolated from the BW5147 murine lymphoblastoid cell line. The initial step in purification was the preparation of a crude plasma membrane fraction followed by acetone precipitation. The acetone pellet was solubilized using deoxycholate (DOC) and Thy-1.1 was purified by use of a Lens culinaris lectin affinity column and an AcA-34 gel filtration column. The purified glycoprotein with Thy-1.1 activity had a mol wt of approximately 25,000 daltons. The isolation of this molecule was effected by detecting Thy-I activity utilizing rabbit anti- mouse brain serum tested on rat thymocytes. Congenic anti-Thy-l.1 serum was ineffective in detecting Thy-l.1 after DOC solubilization. An antiserum prepared in rabbits to the purified Thy-1.1 was found to be cytotoxic to mouse and rat thymocytes. The cytotoxic activity of this antisera could be completely absorbed with AKR/Jax brain and thymus but was not absorbed by liver. In addition, AKR/Jax thymocytes totally absorbed all cytotoxic activity of the rabbit anti-purified Thy-1 serum for BW5147 cells suggesting that the cell line shares identical specificities with normal thymocytes. The purified Thy-1.1 molecule was able to totally absorb the cytotoxic activity of mouse congenic anti-Thy-1. These studies serve as a model for the isolation of other murine lymphoid cell surface components in quantities for detailed structural and functional analysis.
Thy-l.1分子是从BW5147小鼠淋巴母细胞系中分离出来的。纯化的第一步是制备粗制质膜组分，然后进行丙酮沉淀。用脱氧胆酸盐（DOC）溶解丙酮沉淀，并用扁豆凝集素亲和柱和AcA - 34凝胶过滤柱纯化Thy-1.1。具有Thy-1.1活性的纯化糖蛋白的分子量约为25,000道尔顿。该分子的分离是通过利用在大鼠胸腺细胞上检测的兔抗小鼠脑血清来检测Thy-I活性来实现的。同基因抗Thy-l.1血清在DOC溶解后检测Thy-l.1无效。发现用兔制备的针对纯化的Thy-1.1的抗血清对小鼠和大鼠胸腺细胞具有细胞毒性。这种抗血清的细胞毒性活性可以被AKR/Jax脑和胸腺完全吸收，但不能被肝脏吸收。此外，AKR/Jax胸腺细胞完全吸收了兔抗纯化的Thy-1血清对BW5147细胞的所有细胞毒性活性，这表明该细胞系与正常胸腺细胞具有相同的特异性。纯化的Thy-1.1分子能够完全吸收小鼠同基因抗Thy-1的细胞毒性活性。这些研究为大量分离其他小鼠淋巴细胞表面成分以进行详细的结构和功能分析提供了一个模型。
