DISSOCIATION OF TNF-ALPHA CYTOTOXIC AND PROINFLAMMATORY ACTIVITIES BY P55 RECEPTOR-SELECTIVE AND P75 RECEPTOR-SELECTIVE TNF-ALPHA MUTANTS
DISSOCIATION OF TNF-ALPHA CYTOTOXIC AND PROINFLAMMATORY ACTIVITIES BY P55 RECEPTOR-SELECTIVE AND P75 RECEPTOR-SELECTIVE TNF-ALPHA MUTANTS
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DOI：
10.1002/j.1460-2075.1994.tb06327.x
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发表时间：
1994-02-15
期刊：
影响因子：
11.4
通讯作者：
LOPEZ, AF
中科院分区：
文献类型：
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作者：
BARBARA, JAJ;SMITH, WB;LOPEZ, AF
人肿瘤坏死因子（tnf - α）是一种多效性细胞因子，能够在体外和体内杀死哺乳动物肿瘤细胞，并增强白细胞和内皮细胞的促炎活性，后者的作用限制了其作为抗肿瘤药物在人类中的使用。利用具有选择性结合TNF- p55受体（TNFR55）或p75受体（TNFR75）的能力的TNF- α突变体，我们在这里表明，TNF- α的这两种主要活性可以分离。tnfr55选择性突变体（R32W， E146K和R32WS86T）与TNFR75结合不良，在引起人类喉癌衍生细胞系（Hep-2）的细胞毒性和人类白血病细胞系（U937）的细胞停滞方面显示出与野生型TNF相似的效力。然而，这些tnfr55选择性突变体的促炎活性低于野生型TNF。具体来说，tnf - α启动人中性粒细胞产生超氧化物和抗体依赖细胞介导的细胞毒性，血小板活化因子合成和内皮粘附减少了170倍。以人脐静脉内皮细胞（HUVEC）对中性粒细胞的粘附性、e -选择素表达、中性粒细胞转运和IL-8分泌为代表的人内皮细胞功能的激活也降低了高达280倍。另一方面，单独或与tnfr55选择性突变体联合测试的tnfr75选择性突变体D143F，尽管能够在tnfr75转染的PC60细胞中引起细胞因子的产生，但并未刺激这些活性。这些结果表明：(i) TNFR55除了介导抗肿瘤活性外，还介导中性粒细胞和内皮细胞的活化，（ii） TNFR75增强TNFR55在中性粒细胞和内皮细胞活化中的作用，以及（iii）可能构建新的TNF分子，其保留充分的抗肿瘤活性，同时表现出降低的促炎活性。
Human tumour necrosis factor alpha (TNF-alpha) is a pleiotropic cytokine capable of killing mammalian tumour cells in vitro and in vivo, and of enhancing the proinflammatory activity of leucocytes and endothelium, the latter effects limiting its usage as an antitumour agent in humans. Using TNF-alpha mutants with a selective capacity to bind to the TNF p55 receptor (TNFR55) or to the p75 receptor (TNFR75) we show here that these two major activities of TNF-alpha can be dissociated. The TNFR55-selective mutants (R32W, E146K and R32WS86T) which bind poorly to TNFR75 displayed similar potency to wild-type TNF in causing cytotoxicity of a human laryngeal carcinoma-derived cell line (Hep-2) and cytostasis in a human leukaemic cell line (U937). However, these TNFR55-selective mutants exhibited lower proinflammatory activity than wild-type TNF. Specifically, TNF-alpha's priming of human neutrophils for superoxide production and antibody-dependent cell-mediated cytotoxicity, platelet-activating factor synthesis and adhesion to endothelium were reduced by up to 170-fold. Activation of human endothelial cell functions represented by human umbilical venular endothelial cell (HUVEC) adhesiveness for neutrophils, E-selectin expression, neutrophil transmigration and IL-8 secretion were also reduced by up to 280-fold. On the other hand, D143F, a TNFR75-selective mutant tested either alone or in combination with TNFR55-selective mutants, did not stimulate these activities despite being able to cause cytokine production in TNFR75-transfected PC60 cells. These results demonstrate that (i) TNFR55, in addition to mediating antitumour activity, also mediates neutrophil and endothelial cell activation, (ii) TNFR75 potentiates the role of TNFR55 in neutrophil and endothelial cell activation, and (iii) novel TNF molecules may be constructed which retain full antitumour activity whilst exhibiting reduced proinflammatory activities.