Department of Protein Engineering, VLST Corporation, Seattle, Washington, United States of America.
PLoS One. 2012;7(8):e43332. doi: 10.1371/journal.pone.0043332. Epub 2012 Aug 17.
Chemokines play a key role in leukocyte recruitment during inflammation and are implicated in the pathogenesis of a number of autoimmune diseases. As such, inhibiting chemokine signaling has been of keen interest for the development of therapeutic agents. This endeavor, however, has been hampered due to complexities in the chemokine system. Many chemokines have been shown to signal through multiple receptors and, conversely, most chemokine receptors bind to more than one chemokine. One approach to overcoming this complexity is to develop a single therapeutic agent that binds and inactivates multiple chemokines, similar to an immune evasion strategy utilized by a number of viruses. Here, we describe the development and characterization of a novel therapeutic antibody that targets a subset of human CC chemokines, specifically CCL3, CCL4, and CCL5, involved in chronic inflammatory diseases. Using a sequential immunization approach, followed by humanization and phage display affinity maturation, a therapeutic antibody was developed that displays high binding affinity towards the three targeted chemokines. In vitro, this antibody potently inhibits chemotaxis and chemokine-mediated signaling through CCR1 and CCR5, primary chemokine receptors for the targeted chemokines. Furthermore, we have demonstrated in vivo efficacy of the antibody in a SCID-hu mouse model of skin leukocyte migration, thus confirming its potential as a novel therapeutic chemokine antagonist. We anticipate that this antibody will have broad therapeutic utility in the treatment of a number of autoimmune diseases due to its ability to simultaneously neutralize multiple chemokines implicated in disease pathogenesis.
趋化因子在炎症期间白细胞募集中发挥关键作用,并与许多自身免疫性疾病的发病机制有关。因此,抑制趋化因子信号转导一直是开发治疗药物的热点。然而,由于趋化因子系统的复杂性,这一努力受到了阻碍。许多趋化因子已被证明通过多种受体信号传递,相反,大多数趋化因子受体结合一种以上的趋化因子。克服这种复杂性的一种方法是开发一种能够结合并失活多种趋化因子的单一治疗性抗体,类似于许多病毒利用的免疫逃避策略。在这里,我们描述了一种新型治疗性抗体的开发和表征,该抗体针对一组人类 CC 趋化因子,特别是 CCL3、CCL4 和 CCL5,这些趋化因子与慢性炎症性疾病有关。通过顺序免疫接种方法,然后进行人源化和噬菌体展示亲和力成熟,开发了一种对三种靶向趋化因子具有高结合亲和力的治疗性抗体。在体外,该抗体可强烈抑制趋化性和趋化因子介导的通过 CCR1 和 CCR5 的信号转导,CCR1 和 CCR5 是靶向趋化因子的主要趋化因子受体。此外,我们在 SCID-hu 小鼠皮肤白细胞迁移模型中证明了该抗体的体内疗效,从而证实了其作为新型治疗性趋化因子拮抗剂的潜力。我们预计,由于该抗体能够同时中和多种与疾病发病机制相关的趋化因子,因此它将在治疗许多自身免疫性疾病方面具有广泛的治疗用途。