CCL5 (formerly RANTES) belongs to the CC (or β) chemokine family and is associated with a plethora of inflammatory disorders and pathologic states. CCL5 is mainly produced and secreted by T cells, macrophages, epithelial cells, and fibroblasts and acts as a chemoattractant to recruit effector cells to the inflammation sites. Chicken CCL5 (chCCL5) protein is closely related to avian CCL5 orthologs but distinct from mammalian orthologs, and its modulatory roles in the immune response are largely unknown. The present work was undertaken to characterize the immunological properties of chCCL5 using the new sets of anti-chCCL5 mouse monoclonal antibodies (mAbs). Eight different mAbs (6E11, 6H1, 8H11, 11G1, 11G11, 12H1, 13D1, and 13G3) were characterized for their specificity and binding ability toward chCCL5. Two (13G3 and 6E11) of them were selected to detect native chCCL5 in chCCL5-specific antigen-capture ELISA. Using 13G3 and 6E11 as capture and detection antibodies, respectively, the ELISA system detected serum chCCL5 secretions in Clostridium perfringens- and Eimeria-infected chickens. The intracellular expressions of chCCL5 in primary cells or cell lines derived from chickens were validated in immunocytochemistry and flow cytometry assays using both 13G3 and 6E11 mAbs. Furthermore, 6E11, but not 13G3, neutralized chCCL5-induced chemotaxis in vitro using chicken PBMCs. These molecular characteristics of chCCL5 demonstrate the potential application of anti-chCCL5 mAbs and CCL5-specific antigen-capture detection ELISA for detecting native chCCL5 in biological samples. The availability of these new immunological tools will be valuable for fundamental and applied studies in avian species.

CCL5（以前称为 RANTES）属于 CC（或 β）趋化因子家族，与大量炎症性疾病和病理状态相关。CCL5 主要由 T 细胞、巨噬细胞、上皮细胞和成纤维细胞产生和分泌，并作为化学引诱剂将效应细胞募集到炎症部位。鸡 CCL5 (chCCL5) 蛋白与禽类 CCL5 直系同源物密切相关，但与哺乳动物直系同源物不同，其在免疫反应中的调节作用在很大程度上是未知的。目前的工作是使用新的抗 chCCL5 小鼠单克隆抗体 (mAb) 组来表征 chCCL5 的免疫学特性。对八种不同的 mAb（6E11、6H1、8H11、11G1、11G11、12H1、13D1 和 13G3）的特异性和对 chCCL5 的结合能力进行了表征。选择其中两个（13G3 和 6E11）在 chCCL5 特异性抗原捕获 ELISA 中检测天然 chCCL5。ELISA 系统分别以 13G3 和 6E11 作为捕获抗体和检测抗体，检测血清 chCCL5 分泌物产气荚膜梭菌和艾美球菌感染的鸡。使用 13G3 和 6E11 mAb 在免疫细胞化学和流式细胞术测定中验证了 chCCL5 在源自鸡的原代细胞或细胞系中的细胞内表达。此外，6E11，但不是 13G3，在体外使用鸡 PBMC中和了 chCCL5 诱导的趋化性。chCCL5 的这些分子特征证明了抗 chCCL5 mAb 和 CCL5 特异性抗原捕获检测 ELISA 在检测生物样品中天然 chCCL5 的潜在应用。这些新的免疫学工具的可用性对于鸟类物种的基础和应用研究将是有价值的。