CHARACTERIZATION OF HUMAN-TUMOR NECROSIS FACTOR PRODUCED BY PERIPHERAL-BLOOD MONOCYTES AND ITS SEPARATION FROM LYMPHOTOXIN
CHARACTERIZATION OF HUMAN-TUMOR NECROSIS FACTOR PRODUCED BY PERIPHERAL-BLOOD MONOCYTES AND ITS SEPARATION FROM LYMPHOTOXIN
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DOI：
10.1002/ijc.2910360112
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发表时间：
1985-01-01
影响因子：
6.4
通讯作者：
VILCEK, J
中科院分区：
文献类型：
--
作者：
KELKER, HC;OPPENHEIM, JD;VILCEK, J
用植物血凝素（PHA）和佛波酯12-0-十四烷酰佛波酯13-乙酸酯（TPA）诱导人外周血白细胞（PBL）培养物产生两种细胞毒蛋白，在以小鼠L 929细胞为靶细胞的体外试验中，这两种蛋白不能区分。其中一种蛋白质具有光毒素（LT）的抗原性和理化性质。另一种细胞毒素被鉴定为肿瘤坏死因子（TNF），主要是根据与TNF抗血清的抗原交叉反应性，以及其特征性的理化性质和细胞来源。与LT不同，PBL衍生的TNF不与伴刀豆球蛋白A-Sepharose或其他几种特异于糖蛋白中常见的碳水化合物部分的凝集素-Sepharose柱结合。通过凝胶过滤测定的天然TNF的MW约为40，000，而SDS-PAGE显示16，500 ± 1的单个尖峰。500.当用PHA和/或TPA刺激通过淘洗分离的单核细胞和淋巴细胞的培养物时，单核细胞是TNF的主要来源。相反，只有淋巴细胞产生LT。TNF和LT的抗血清的混合物中和了所有的细胞毒性的粗品人淋巴因子制剂的L 929细胞，这表明TNF和LT是唯一的，或主要的，细胞毒性蛋白存在于这样的粗品淋巴因子制剂证明在本试验中。
Cultures of human peripheral blood leukocytes (PBL) induced with phytohemagglutinin (PHA) and the phorbol ester 12-0-tetradecanoylphorbol 13-acetate (TPA) produced 2 types of cytotoxic proteins, indistinguishable in the in vitro assay employing murine L 929 cells as targets. One of these proteins had the antigenic and physicochemical properties of lymphotoxin (LT). The other cytotoxin was identified as tumor necrosis factor (TNF), mainly on the basis of antigenic cross-rectivity demonstrated with antiserum to TNF, and also by its characteristic physicochemical properties and cell source. Unlike LT, PBL-derived TNF did not bind to Concanavalin A-Sepharose or to several other agglutinin-Sepharose columns specific for carbohydrate moieties common in glycoproteins. The MW of native TNF determined by gel filtration was approximately 40,000 while SDS-PAGE revealed a single sharp peak of 16,500 .+-. 500. When cultures of monocytes and lymphocytes separated by elutriation were stimulated with PHA and/or TPA, monocytes were the major source of TNF. In contrast, only lymphocytes produced LT. A mixture of antisera to TNF and LT neutralized all cytoxicity of crude human lymphokine preparations for L 929 cells, suggesting that TNF and LT are either the only, or the major, cytotoxic proteins present in such crude lymphokine preparations demonstrable in this assay.