Augmentation of spontaneous macrophage-mediated cytolysis by eosinophil peroxidase.
Augmentation of spontaneous macrophage-mediated cytolysis by eosinophil peroxidase.
复制标题
DOI：
10.1084/jem.155.5.1291
复制
发表时间：
1982-05-01
期刊：
影响因子：
--
通讯作者：
Klebanoff SJ
中科院分区：
文献类型：
--
作者：
Nathan CF;Klebanoff SJ
嗜酸性粒细胞过氧化物酶（EPO）是一种从马血中纯化的阳离子蛋白质，它粘附在四种不同类型的肿瘤细胞上，通过预先形成或酶促产生的H（2）O（2）显著增强它们的溶解作用（在不含补充卤化物的含血清组织培养基中测定，可达76倍）。同样，与未涂覆的肿瘤细胞相比，EPO涂覆的肿瘤细胞与PMA触发呼吸爆发的巨噬细胞或粒细胞一起孵育时，对溶解的敏感性高达32倍。然而，在不存在外源性触发剂的情况下，EPO包被的肿瘤细胞也容易被卡介苗激活的巨噬细胞裂解。这种自发的细胞溶解是快速的（在2小时时50%）和有效的（在巨噬细胞/肿瘤细胞比率为1.5至4.6时50%溶解），并且在过氧化物敏感性肿瘤（TLX 9）和过氧化物抗性肿瘤（NK淋巴瘤）中均观察到。在所使用的条件下，单独的EPO和单独的巨噬细胞都不能自发地溶细胞。EPO和EPO包被的肿瘤细胞都没有引发巨噬细胞释放H（2）0（2）的可检测增量。尽管如此，巨噬细胞介导的EPO包被的肿瘤细胞的自发性细胞溶解完全可被过氧化氢酶（50%抑制，23 U/ml）抑制，尽管不能被加热的过氧化氢酶抑制，表明需要H（2）0（2）。叠氮化物（50%抑制，2.6 × 10 - 5 M）也可完全抑制细胞溶解，表明EPO的酶活性是必需的。因此，嗜酸性粒细胞的嗜细胞过氧化物酶和活化的巨噬细胞自发释放的H（2）0（2）在生理介质中协同作用，破坏肿瘤细胞。
Eosinophil peroxidase (EPO), a cationic protein purified from horse blood, adhered to four different types of tumor cells, markedly potentiating their lysis by preformed or enzymatically generated H(2)0(2) (up to 76-fold, as assayed in serum-containing tissue culture medium without supplemental halide). Similarly, compared with uncoated tumor cells, EPO-coated tumor cells were up to 32 times more sensitive to lysis when incubated with macrophages or granulocytes whose respiratory burst was triggered by PMA. However, EPO-coated tumor cells were also readily lysed by bacillus Calmette- Guerin-activated macrophages in the absence of exogenous triggering agents. This spontaneous cytolysis was rapid (50 percent at 2 h) and potent (50 percent lysis at macrophage/tumor cell ratios of 1.5 to 4.6), and was observed with both a peroxide-sensitive tumor (TLX9) and a peroxide-resistant tumor (NK lymphoma). Under the conditions used, neither EPO alone nor macrophages alone were spontaneously cytolytic. Neither EPO nor EPO-coated tumor cells triggered a detectable increment in H(2)0(2) release from macrophages. Nonetheless, spontaneous macrophage-mediated cytolysis of EPO- coated tumor cells was completely inhibitable by catalase (50 percent inhibition, 23 U/ml), although not by heated catalase, indicating a requirement for H(2)0(2). Cytolysis was also completely inhibitable by azide (50 percent inhibition, 2.6 X 10 -5 M), indicating a requirement for enzymatic activity of EPO. Thus, a cytophilic peroxidase from eosinophils and H(2)0(2) spontaneously released from activated macrophages interacted synergistically in a physiologic medium to destroy tumor cells.