Neutralization of leukotriene C4 and D4 activity by monoclonal and single-chain antibodies.
Neutralization of leukotriene C4 and D4 activity by monoclonal and single-chain antibodies.
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通过单克隆和单链抗体中和白三烯 C4 和 D4 活性。
DOI：
10.1016/j.bbagen.2013.12.016
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Takahashi Y.
中科院分区：
文献类型：
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作者：
Kawakami Y;Hirano S;Kinoshita M;Otsuki A;Suzuki-Yamamoto T; Suzuki M;Kimoto M;Sasabe S;Fukushima M;Kishimoto K;Izumi T;Oga T;Narumiya S;Sugahara M;Miyano M;Yamamoto S;Takahashi Y.
背景半胱氨酰白三烯（LT）是炎症的关键介质。为了探索LTC4单克隆抗体（mAbLTC）的抗原识别位点结构，我们之前分离了抗体重链和轻链的全长cDNA，并制备了包含这两条链可变区的单链抗体（scFvLTC）。方法我们检查mAbLTC和scFvLTC是否通过竞争与其受体的结合来中和LTC4和LTD4的生物活性。结果mAbLTC和scFvLTC抑制 LTC4 或 LTD4 与中国仓鼠卵巢细胞中过表达的 CysLT1 受体 (CysLT1R) 和 CysLT2 受体 (CysLT2R) 的结合。在表达 CysLT1R 的人单核细胞白血病 THP-1 细胞中，LTD4 对单核细胞趋化蛋白 1 和白细胞介素 8 mRNA 的诱导不仅受到 mAbLTC 的剂量依赖性抑制，而且还受到 scFvLTC 的剂量依赖性抑制。 mAbLTC 或 scFvLTC 剂量依赖性地抑制 LTC4 和 LTD4 诱导的表达 CysLT2R 的小鼠血小板聚集。在小鼠哮喘模型中观察到，给予 mAbLTC 可以减少肺部嗜酸性粒细胞浸润和杯状细胞增生。此外，mAbLTC 与 CysLT2R 拮抗剂结合，但不与 CysLT1R 拮抗剂结合。结论这些结果表明，mAbLTC 和 scFvLTC 通过竞争 LT 与 CysLT1R 和 CysLT2R 的结合来中和 LT 的生物活性。此外，半胱氨酰LT受体拮抗剂与mAbLTC的结合表明该抗体的LT识别位点与这些受体的LT识别位点结构相似。一般意义mAbLTC可用于治疗炎症性疾病，例如哮喘。
BackgroundCysteinyl leukotrienes (LTs) are key mediators in inflammation. To explore the structure of the antigen-recognition site of a monoclonal antibody against LTC4(mAbLTC), we previously isolated full-length cDNAs for heavy and light chains of the antibody and prepared a single-chain antibody comprising variable regions of these two chains (scFvLTC).MethodsWe examined whether mAbLTC and scFvLTC neutralized the biological activities of LTC4and LTD4by competing their binding to their receptors.ResultsmAbLTC and scFvLTC inhibited their binding of LTC4or LTD4to CysLT1receptor (CysLT1R) and CysLT2receptor (CysLT2R) overexpressed in Chinese hamster ovary cells. The induction by LTD4of monocyte chemoattractant protein-1 and interleukin-8 mRNAs in human monocytic leukemia THP-1 cells expressing CysLT1R was dose-dependently suppressed not only by mAbLTC but also by scFvLTC. LTC4- and LTD4-induced aggregation of mouse platelets expressing CysLT2R was dose-dependently suppressed by either mAbLTC or scFvLTC. Administration of mAbLTC reduced pulmonary eosinophil infiltration and goblet cell hyperplasia observed in a murine model of asthma. Furthermore, mAbLTC bound to CysLT2R antagonists but not to CysLT1R antagonists.ConclusionsThese results indicate that mAbLTC and scFvLTC neutralize the biological activities of LTs by competing their binding to CysLT1R and CysLT2R. Furthermore, the binding of cysteinyl LT receptor antagonists to mAbLTC suggests the structural resemblance of the LT-recognition site of the antibody to that of these receptors.General significancemAbLTC can be used in the treatment of inflammatory diseases such as asthma.