Wang Junxiang, Wang Lu, Xiang Ying, Ricklin Daniel, Lambris John D, Chen Gang
Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Institute of Organ Transplantation, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; Key Laboratory of Organ Transplantation, Ministry of Education, Wuhan, China; Key Laboratory of Organ Transplantation, Ministry of Public Health, Wuhan, China.
Clin Immunol. 2016 Jan;162:37-44. doi: 10.1016/j.clim.2015.11.002. Epub 2015 Nov 6.
Simple and reliable methods for evaluating the inhibitory effects of drug candidates on complement activation are essential for preclinical development. Here, using an immortalized porcine aortic endothelial cell line (iPEC) as target, we evaluated the feasibility and effectiveness of an in vitro xenoantibody-mediated complement-dependent cytotoxicity (CDC) model for evaluating the complement inhibitory activity of Cp40, a potent analog of the peptidic C3 inhibitor compstatin. The binding of human xenoantibodies to iPECs led to serum dilution-dependent cell death. Pretreatment of the human serum with Cp40 almost completely inhibited the deposition of C3 fragments and C5b-9 on the cells, resulting in a dose-dependent inhibition of CDC against the iPECs. Using the same method to compare the effects of Cp40 on complement activation in humans, rhesus and cynomolgus monkeys, we found that the inhibitory patterns were similar overall. Thus, the in vitro xenoantibody-mediated CDC assay may have considerable potential for future clinical use.
在临床前开发中，用于评估候选药物对补体激活抑制作用的简单可靠方法至关重要。在此，我们以永生化猪主动脉内皮细胞系（iPEC）为靶点，评估了一种体外异种抗体介导的补体依赖性细胞毒性（CDC）模型用于评估肽类C3抑制剂compstatin的强效类似物Cp40补体抑制活性的可行性和有效性。人异种抗体与iPEC的结合导致了血清稀释依赖性细胞死亡。用Cp40预处理人血清几乎完全抑制了C3片段和C5b-9在细胞上的沉积，从而导致对iPEC的CDC呈剂量依赖性抑制。使用相同方法比较Cp40对人、恒河猴和食蟹猴补体激活的影响，我们发现总体抑制模式相似。因此，体外异种抗体介导的CDC检测在未来临床应用中可能具有相当大的潜力。
