Dorgham Karim, Dejou Cécile, Piesse Christophe, Gorochov Guy, Pène Jérôme, Yssel Hans
Sorbonne Universités, Inserm U1135, UPMC Univ Paris 06 UMRS 1135, Centre d'Immunologie et des Maladies Infectieuses (Cimi-Paris), F-75013, Paris, France.
Institut de Recherche en Cancérologie, F-34090, Montpellier, France.
PLoS One. 2016 Jun 23;11(6):e0157740. doi: 10.1371/journal.pone.0157740. eCollection 2016.
Chemokines and their receptors play an important role in cell trafficking and recruitment. The CCR6 chemokine receptor, selectively expressed on leukocyte populations, has been shown to play a deleterious role in the pathogenesis of various chronic inflammatory diseases and, as such, may constitute a prime target in the development of immunotherapeutic treatment. However, to date no neutralizing mouse monoclonal antibodies (mAbs) specific for this chemokine receptor have been reported, whereas information on small molecules capable of interfering with the interaction of CCR6 and its ligands is scant. Here, we report the failure to generate neutralizing mouse mAbs specific for human (hu)CCR6. Immunization of mice with peptides mimicking extracellular domains, potentially involved in CCR6 function, failed to induce Abs reactive with the native receptor. Although the use of NIH-3T3 cells expressing huCCR6 resulted in the isolation of mAbs specific for this receptor, they were not able to block the interaction between huCCR6 and huCCL20. Investigation of the anti-huCCR6 mAbs generated in the present study, as well as those commercially available, show that all mAbs invariably recognize a unique, non-neutralizing, immunodominant region in the first part of its N-terminal domain. Together, these results indicate that the generation of potential neutralizing anti-huCCR6 mAbs in the mouse is unlikely to succeed and that alternative techniques, such as the use of other animal species for immunization, might constitute a better approach to generate such a potentially therapeutic tool for the treatment of inflammatory disease.
趋化因子及其受体在细胞迁移和募集过程中发挥着重要作用。CCR6趋化因子受体在白细胞群体中选择性表达，已证实在多种慢性炎症疾病的发病机制中起有害作用，因此可能成为免疫治疗开发的主要靶点。然而，迄今为止，尚未报道针对该趋化因子受体的中和小鼠单克隆抗体（mAb），而关于能够干扰CCR6及其配体相互作用的小分子的信息也很少。在此，我们报告未能产生针对人（hu）CCR6的中和小鼠mAb。用模拟可能参与CCR6功能的细胞外结构域的肽免疫小鼠，未能诱导出与天然受体反应的抗体。尽管使用表达huCCR6的NIH-3T3细胞分离出了针对该受体的mAb，但它们无法阻断huCCR6与huCCL20之间的相互作用。对本研究中产生的以及市售的抗huCCR6 mAb的研究表明，所有mAb均始终识别其N端结构域第一部分中的一个独特的、非中和性的免疫显性区域。总之，这些结果表明在小鼠中产生潜在的中和抗huCCR6 mAb不太可能成功，而其他技术，如使用其他动物物种进行免疫，可能是生成这种潜在治疗炎症性疾病工具的更好方法。
