Generation of antagonistic monoclonal antibodies against the neoepitope of active mouse interleukin (IL)-18 cleaved by inflammatory caspases
Generation of antagonistic monoclonal antibodies against the neoepitope of active mouse interleukin (IL)-18 cleaved by inflammatory caspases
复制标题
产生针对由炎性半胱天冬酶裂解的活性小鼠白细胞介素 (IL)-18 新表位的拮抗性单克隆抗体
DOI：
10.1016/j.abb.2022.109322
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发表时间：
2022
影响因子：
3.9
通讯作者：
Kamino Hiroki
中科院分区：
文献类型：
--
作者：
Uchida Yuki;Nariai Yuko;Obayashi Eiji;Tajima Yoshitsugu;Koga Tomohiro;Kawakami Atsushi;Urano Takeshi;Kamino Hiroki
白细胞介素18 （Interleukin 18, IL-18）是IL-1家族的一员，在先天和获得性免疫系统中都起着重要的作用。它在各种细胞类型中组成性表达为无活性前体（24 kDa），由炎性caspase-1/4切割的成熟IL-18 （18 kDa）与白细胞介素-18受体结合，从而激活下游信号通路。我们之前生成的抗人IL-18抗体特异性识别由炎性caspase-1/4切割的人IL-18新表位。由于人IL-18和小鼠IL-18新表位的n端氨基酸序列不同，抗人IL-18新表位抗体不能识别小鼠成熟IL-18。我们现在已经产生了新的抗小鼠IL-18新表位抗体。我们还证实了通过小鼠IL-18刺激P-815小鼠细胞分泌CXCL2，并建立了一种简单的方法来评估小鼠IL-18的活性。利用该评价体系，我们证实了抗小鼠IL-18新表位抗体能够抑制小鼠IL-18。通过在小鼠模型中证明本研究建立的抗小鼠IL-18新表位和功能阻断单抗的治疗效果，对应于IL-18可能参与的人类炎症性疾病，如炎症性肠病，我们可以提供先前建立的抗人IL-18新表位和功能阻断单抗在对应于小鼠模型的人类炎症性疾病中的作用的概念证明。
Interleukin 18 (IL-18) is a member of the IL-1 family and plays an important role in both the innate and acquired immune systems. It is constitutively expressed as an inactive precursor (24 kDa) in various cell types, and the mature IL-18 (18 kDa) cleaved by inflammatory caspase-1/4 binds to the interleukin-18 receptor, thereby activating downstream signaling pathways. We previously generated anti-human IL-18 antibodies that specifically recognize the human IL-18 neoepitope cleaved by inflammatory caspase-1/4. Because the N-terminal amino acid sequences of the neoepitopes are different between human IL-18 and mouse IL-18, the anti-human IL-18 neoepitope antibodies do not recognize mouse mature IL-18. We have now generated novel anti-mouse IL-18 neoepitope antibodies. We also confirmed CXCL2 secretion from P-815 mouse cells by mouse IL-18 stimulation, and established a simple assay to evaluate the activity of mouse IL-18. Using this evaluation system, we confirmed that the anti-mouse IL-18 neoepitope antibodies could inhibit mouse IL-18. By demonstrating the therapeutic efficacy of the anti-mouse IL-18 neoepitope and function-blocking mAbs established in the present study in mouse models, corresponding to human inflammatory diseases in which IL-18 may be involved, such as inflammatory bowel diseases, we can provide the proof-of-concept that the previously established anti-human IL-18 neoepitope and function-blocking mAbs work in human inflammatory disorders corresponding to mouse models.