Both CC and CXC-class chemokines drive inflammatory disease. Tick salivary chemokine-binding proteins (CKBPs), or evasins, specifically bind subsets of CC- or CXC-chemokines, and could precisely target disease-relevant chemokines. Here we have used yeast surface display to identify two tick evasins: a CC-CKBP, P1243 from Amblyomma americanum and a CXC-CKBP, P1156 from Ixodes ricinus. P1243 binds 11 CC-chemokines with K_d < 10 nM, and 10 CC-chemokines with K_d between 10 and 100 nM. P1156 binds two ELR + CXC-chemokines with K_d < 10 nM, and four ELR + CXC-chemokines with K_d between 10 and 100 nM. Both CKBPs neutralize chemokine activity with IC₅₀ < 10 nM in cell migration assays. As both CC- and CXC-CKBP activities are desirable in a single agent, we have engineered "two-warhead" CKBPs to create single agents that bind and neutralize subsets of CC and CXC chemokines. These results show that tick evasins can be linked to create non-natural proteins that target subsets of CC and CXC chemokines. We suggest that "two-warhead" evasins, designed by matching the activities of parental evasins to CC and CXC chemokines expressed in disease, would achieve precision targeting of inflammatory disease-relevant chemokines by a single agent.

中文翻译：

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基因工程双弹头evasin提供了一种精确靶向疾病相关趋化因子子集的方法。

CC 和 CXC 类趋化因子都会导致炎症性疾病。蜱唾液趋化因子结合蛋白 (CKBP) 或 evasins 特异性结合 CC 或 CXC 趋化因子子集，并且可以精确地靶向与疾病相关的趋化因子。在这里，我们使用酵母表面展示来识别两种蜱逃避素：来自美洲钝蜱的CC-CKBP，P1243和来自蓖麻硬蜱的CXC-CKBP，P1156。P1243 结合 11 个 K _d  < 10 nM 的 CC 趋化因子，以及 10 个 K _d介于 10 和 100 nM 之间的 CC 趋化因子。P1156 结合两种 ELR + CXC 趋化因子，K _d  < 10 nM，并结合四种 ELR + CXC 趋化因子，K _d在 10 至 100 nM 之间。 在细胞迁移测定中，两种 CKBP 均能中和趋化因子活性，IC _{50 < 10 nM。}由于 CC- 和 CXC-CKBP 活性在单一药剂中是理想的，因此我们设计了“双弹头”CKBP 来创建结合并中和 CC 和 CXC 趋化因子子集的单一药剂。这些结果表明，蜱逃避蛋白可以连接起来产生针对 CC 和 CXC 趋化因子子集的非天然蛋白质。我们认为，通过将亲代evasins的活性与疾病中表达的CC和CXC趋化因子相匹配而设计的“双弹头”evasins将通过单一药物实现炎症性疾病相关趋化因子的精确靶向。