Complement-mediated cell death induced by rituximab in B-cell lymphoproliferative disorders is mediated in vitro by a caspase-independent mechanism involving the generation of reactive oxygen species
Complement-mediated cell death induced by rituximab in B-cell lymphoproliferative disorders is mediated in vitro by a caspase-independent mechanism involving the generation of reactive oxygen species
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DOI：
10.1182/blood.v98.9.2771
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发表时间：
2001-11-01
期刊：
影响因子：
20.3
通讯作者：
Montserrat, E
中科院分区：
文献类型：
--
作者：
Bellosillo, B;Villamor, N;Montserrat, E
对利妥昔单抗在55例B细胞淋巴增殖性疾病患者细胞中的体外作用机制进行了研究。当细胞仅与利妥昔单抗一起孵育时，未观察到细胞毒性作用，但在有人AB血清存在的情况下，利妥昔单抗诱导补体依赖性细胞死亡（R - CDC）。在33例B细胞慢性淋巴细胞白血病患者中的9例、16例套细胞淋巴瘤患者中的16例、4例滤泡性淋巴瘤患者中的4例以及2例毛细胞白血病患者中的2例的细胞中观察到细胞毒性作用。在每个细胞表达超过50×10³个CD20分子的患者细胞中观察到R - CDC，且与每个细胞的CD20分子数量直接相关。用抗 - CD59预孵育增加了利妥昔单抗的细胞毒性作用，并使非敏感病例的细胞敏感化。在R - CDC中既未观察到多聚ADP核糖聚合酶（PARP）的裂解，也未观察到半胱天冬酶 - 3的激活。此外，未检测到具有亚二倍体DNA含量的细胞，并且一种广谱半胱天冬酶抑制剂不能阻止R - CDC，这表明是一种不依赖半胱天冬酶的机制。在AB血清存在的情况下与利妥昔单抗一起孵育会诱导活性氧物质（ROS）快速且大量产生。通过用两种ROS清除剂N - 乙酰 - L - 半胱氨酸（NAC）或Tiron孵育细胞可阻断R - CDC，这表明细胞毒性作用是由于超氧阴离子（O₂⁻）自由基的产生。总之，本研究的结果表明，CD20、CD59和补体在利妥昔单抗的体外细胞毒性作用中起作用，这种作用是由一种涉及ROS产生的不依赖半胱天冬酶的过程所介导的。（C）2001年美国血液学会版权所有
Mechanisms involving the in vitro effect of rituximab in cells from 55 patients with B-cell lymphoproliferative disorders were investigated. No cytotoxic effect was observed when cells were incubated with rituximab alone, but in the presence of human AB serum rituximab induced complement-dependent cell death (R-CDC). A cytotoxic effect was observed in cells from 9 of 33 patients with B-cell chronic lymphocytic leukemia, 16 of 16 patients with mantle-cell lymphoma, 4 of 4 patients with follicular lymphoma, and 2 of 2 patients with hairy-cell leukemia. R-CDC was observed in cells from patients expressing more than 50 x 10(3) CD20 molecules per cell, and directly correlated with the number of CD20 molecules per cell. Preincubation with anti-CD59 increased the cytotoxic effect of rituximab and sensitized cells from nonsensitive cases. Neither cleavage of poly-ADP ribose polymerase (PARP) nor activation of caspase-3 was observed in R-CDC. In addition, no cells with a hypodiploid DNA content were detected and R-CDC was not prevented by a broad-spectrum caspase inhibitor, suggesting a caspase-independent mechanism. Incubation with rituximab in the presence of AB serum induced a rapid and intense production of reactive oxygen species (ROS). R-CDC was blocked by the incubation of cells with N-acetyl-L-cysteine (NAC) or Tiron, 2 ROS scavengers, indicating that the cytotoxic effect was due to the generation of superoxide (O-2(-)) radicals. In conclusion, the results of the present study suggest that CD20, CD59, and complement have a role in the in vitro cytotoxic effect of rituximab, which is mediated by a caspase-independent process that involves ROS generation. (C) 2001 by The American Society of Hematology.