BIOCHEMICAL AND BIOLOGICAL CHARACTERIZATION OF LYMPHOCYTE REGULATORY MOLECULES .1. PURIFICATION OF A CLASS OF MURINE LYMPHOKINES
BIOCHEMICAL AND BIOLOGICAL CHARACTERIZATION OF LYMPHOCYTE REGULATORY MOLECULES .1. PURIFICATION OF A CLASS OF MURINE LYMPHOKINES
复制标题
DOI：
10.1084/jem.150.4.849
复制
发表时间：
1979-01-01
影响因子：
15.3
通讯作者：
SMITH, KA
中科院分区：
文献类型：
--
作者：
WATSON, J;GILLIS, S;SMITH, KA
小鼠脾细胞经刀豆球蛋白（Con）A激活后，可产生一类淋巴因子分子，在许多淋巴细胞反应试验中具有生物学活性。根据凝胶过滤研究估计，MW为30，000的α-phokines可分解为2种电荷不同的组分，等电点（pI）值分别为4.3和4.9。这两种成分刺激建立的T[胸腺衍生]细胞系在培养中的生长，在Con A存在下的胸腺细胞的增殖，其中Con A单独是非促有丝分裂的培养条件下，在无胸腺（裸）脾培养物中诱导对异源红细胞抗原的抗体应答，在胸腺细胞培养物中产生细胞毒性T淋巴细胞（CTL），在裸脾培养物中产生CTL。在这些培养系统中的每一个中，所述测定显然检测直接作用于活化的T细胞的单一类别的淋巴因子。未活化的T细胞在对淋巴因子产生应答之前必须被抗原或有丝分裂原刺激，但不需要抗原或有丝分裂原与淋巴因子一起继续生长。这两种分子种类，可通过等电聚焦分离，被称为T细胞生长因子（TCGF）。一种大小（30，000道尔顿）与TCGF相似但电荷异质（pI 3.0-4.0）的淋巴因子在T细胞耗尽的脾培养物中刺激对红细胞抗原的免疫应答，但在所述的其他淋巴细胞测定系统中没有刺激活性。小鼠TCGF的两种分子形式（pI 4.3和4.9）显然负责许多淋巴因子活动，如胸腺细胞有丝分裂因子，非特异性T细胞替代因子和杀伤辅助因子或共刺激因子。另一种淋巴因子，通过电荷与TCGF分离，显然具有真正的T细胞替代活性。
Murine spleen cells activated by concanavalin (Con)A in culture produced a class of lymphokine molecules which possess biological activity in a number of lymphocyte response assays. Lymphokines with a MW of 30,000, as estimated from gel filtration studies, could be resolved into 2 components which differ by charge, with isoelectric point (pI) values of 4.3 and 4.9, respectively. Both components stimulated growth of established T[thymus-derived] cell lines in culture, proliferation of thymocytes in the presence of Con A under culture conditions where Con A alone is nonmitogenic, induction of antibody responses to heterologous erythrocyte antigens in athymic (nude) spleen cultures, generation of cytotoxic T lymphocytes (CTL) in thymocyte cultures and generation of CTL in nude spleen cultures. In each of these culture systems the assays were apparently detecting a single class of lymphokine which acts directly on activated T cells. Nonactivated T cells must be stimulated by antigen or mitogen before becoming responsive to lymphokine, but do not require antigen or mitogen for continued growth with lymphokine. The 2 molecular species, separable by isoelectric focusing, were referred to as the T cell growth factor (TCGF). A lymphokine similar in size (30,000 daltons) to TCGF but heterogeneous in charge (pI 3.0-4.0) stimulated immune responses to erythrocyte antigens in T cell-depleted spleen cultures but had no stimulatory activity in other lymphocyte assay systems described. The 2 molecular forms of murine TCGF (pI 4.3 and 4.9) were apparently responsible for many lymphokine activities described elsewhere as thymocyte mitogenic factor, nonspecific T cell-replacing factor and killer helper factor or costimulator. The other lymphokine, separable from TCGF by charge, apparently had true T cell-replacing activity.