Anti-GD(2) with an FC point mutation reduces complement fixation and decreases antibody-induced allodynia.
Anti-GD(2) with an FC point mutation reduces complement fixation and decreases antibody-induced allodynia.
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DOI：
10.1016/j.pain.2010.01.024
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发表时间：
2010-04
期刊：
影响因子：
7.4
通讯作者：
Yu AL
中科院分区：
文献类型：
--
作者：
Sorkin LS;Otto M;Baldwin WM 3rd;Vail E;Gillies SD;Handgretinger R;Barfield RC;Yu HM;Yu AL
针对GD 2神经节苷脂的单克隆抗体，如人-小鼠嵌合抗体ch14.18，已被证明可有效治疗神经母细胞瘤。然而，治疗与全身性、相对阿片类药物耐受性疼痛有关。我们研究了在ch14.18抗体（hu14.18K332A）中限制补体依赖性细胞毒性（CDC）的点突变是否会改善疼痛行为，同时保留抗体依赖性细胞毒性（ADCC）。在体外，CDC和ADCC使用铕-TDA测定。在体内，通过在将ch14.18或hu14.18K332注射到野生型大鼠或具有补体因子6缺陷的大鼠中后测量施加到后爪的von Frey细丝的阈值来评价异常性疼痛。其他大鼠用补体因子C5 a受体拮抗剂预处理，并在注射ch14.18后进行检测。该突变降低了抗体激活补体的能力，同时保持其ADCC能力。注射hu14.18K322（1或3 mg/kg）比注射ch14.18（1 mg/kg）产生的异常性疼痛更快消退。注射ch14.18（1 mg/kg）到C6补体缺乏的大鼠进一步减少抗体诱导的异常性疼痛，而补体因子C5 a受体拮抗剂的预处理完全消除ch14.18诱导的异常性疼痛。这些发现表明，突变体hu14.18 K322引起的异常性疼痛比ch14.18少，并且ch14.18引起的异常性疼痛是由于补体级联的激活：部分地，形成膜攻击复合物，但更重要的是释放补体因子C5 a。开发具有降低的补体依赖性裂解同时维持细胞细胞毒性的免疫抑制剂可以提供具有降低的不良副作用的治疗选择，从而允许治疗性抗体的剂量递增。
Monoclonal antibodies against GD2 ganglioside, such as ch14.18, the human–mouse chimeric antibody, have been shown to be effective for the treatment of neuroblastoma. However, treatment is associated with generalized, relatively opiate-resistant pain. We investigated if a point mutation in ch14.18 antibody (hu14.18K332A) to limit complement-dependent cytotoxicity (CDC) would ameliorate the pain behavior, while preserving antibody-dependent cellular cytotoxicity (ADCC). In vitro, CDC and ADCC were measured using europium-TDA assay. In vivo, allodynia was evaluated by measuring thresholds to von Frey filaments applied to the hindpaws after injection of either ch14.18 or hu14.18K332 into wild type rats or rats with deficient complement factor 6. Other rats were pretreated with complement factor C5a receptor antagonist and tested following ch14.18 injection. The mutation reduces the antibody’s ability to activate complement, while maintaining its ADCC capabilities. Injection of hu14.18K322 (1 or 3 mg/kg) produced faster resolving allodynia than that engendered by ch14.18 (1 mg/kg). Injection of ch14.18 (1 mg/kg) into rats with C6 complement deficiency further reduced antibody-induced allodynia, while pre-treatment with complement factor C5a receptor antagonist completely abolished ch14.18-induced allodynia. These findings showed that mutant hu14.18 K322 elicited less allodynia than ch14.18 and that ch14.18-elicited allodynia is due to activation of the complement cascade: in part, to formation of membrane attack complex, but more importantly to release of complement factor C5a. Development of immunotherapeutic agents with decreased complement-dependent lysis while maintaining cellular cytotoxicity may offer treatment options with reduced adverse side effects, thereby allowing dose escalation of therapeutic antibodies.
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