ADAM10 inhibition of human CD30 shedding increases specificity of targeted immunotherapy in vitro
ADAM10 inhibition of human CD30 shedding increases specificity of targeted immunotherapy in vitro
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DOI：
10.1158/0008-5472.can-06-2470
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发表时间：
2007-01-01
期刊：
影响因子：
11.2
通讯作者：
Hansen, Hinrich P.
中科院分区：
文献类型：
--
作者：
Eichenauer, Dennis A.;Simhadri, Vijaya Lakshmi;Hansen, Hinrich P.
CD30是一种在许多人淋巴瘤细胞上选择性过表达的跨膜蛋白，因此是基于抗体的免疫治疗的一个有趣靶点。然而，治疗性抗体的结合刺激CD30的近膜切割，导致靶抗原的丢失和CD30可溶性外结构域（sCD30）的释放增强。在这里，我们发现sCD30结合CD30配体（CD153）表达的非靶细胞。由于抗体与sCD30结合，这导致不想要的抗体通过sCD30桥接与这些细胞结合。为了克服cd30特异性免疫治疗中正常细胞的脱落依赖性损伤，我们分析了cd30释放的机制。CD30的脱落可以通过蛋白激酶C （PKC）的激活而增强，这涉及到崩解素金属蛋白酶ADAM17，而不是释放胞质钙。然而，抗体诱导的CD30脱落是钙依赖和PKC独立的。这种脱落涉及相关的金属蛋白酶ADAM10，如使用优先的ADAM10抑制剂GI254023X和从胚胎致死性ADAM10（-/-）小鼠产生的ADAM10缺陷细胞系所示。在共培养实验中，抗体诱导的sCD30从人霍奇金淋巴瘤细胞系L540向cd30阴性但cd153表达的人肥大细胞系HMC-1转移被GI254023X抑制。这些发现表明，选择性金属蛋白酶抑制剂阻断抗体诱导的靶抗原脱落可能具有治疗价值，可以提高单克隆抗体免疫治疗的特异性并减少副作用。
CD30 is a transmembrane protein selectively overexpressed on many human lymphoma cells and therefore an interesting target for antibody-based immunotherapy. However, binding of therapeutic antibodies stimulates a juxtamembrane cleavage of CD30 leading to a loss of target antigen and an enhanced release of the soluble ectodomain of CD30 (sCD30). Here, we show that sCD30 binds to CD30 ligand (CD153)-expressing non-target cells. Because antibodies bind to sCD30, this results in unwanted antibody binding to these cells via sCD30 bridging. To overcome shedding-dependent damage of normal cells in CD30-specific immunotherapy, we analyzed the mechanism involved in the release. Shedding of CD30 can be enhanced by protein kinase C (PKC) activation, implicating the disintegrin metalloproteinase ADAM17 but not free cytoplasmic calcium. However, antibody-induced CD30 shedding is calcium dependent and PKC independent. This shedding involved the related metalloproteinase ADAM10 as shown by the use of the preferential ADAM10 inhibitor GI254023X and by an ADAM10-deficient cell line generated from embryonically lethal ADAM10(-/-) mouse. In coculture experiments, the antibody-induced transfer of sCD30 from the human Hodgkin's lymphoma cell line L540 to the CD30-negative but CD153-expressing human mast cell line HMC-1 was inhibited by GI254023X. These findings suggest that selective metalloproteinase inhibitors blocking antibody-induced shedding of target antigens could be of therapeutic value to increase the specificity and reduce side effects of immunotherapy with monoclonal antibodies.