Kim Woo H, Lillehoj Hyun S, Lim Yeaseul, Min Wongi, Sullivan Yvonne B, Kakach Laura, LaBresh Joanna W
Animal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, ARS, U.S. Department of Agriculture, Beltsville, MD 20705, USA.
Animal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, ARS, U.S. Department of Agriculture, Beltsville, MD 20705, USA.
Dev Comp Immunol. 2017 Jul;72:30-36. doi: 10.1016/j.dci.2017.02.008. Epub 2017 Feb 20.
Interleukin-8(IL-8)/CXCL8 is a CXC-family chemokine that attracts lymphocytes to sites of tissue damage and plays a role in the inflammatory response and wound healing. Chicken chemotactic and angiogenic factor was referred to as chCXCLi2 and has been studied as one of human CXCL8 homologue for more than 20 years. However, no monoclonal antibodies (mAbs) that specifically detect chCXCLi2 have been developed. Here, we developed and characterized mouse mAbs against chCXCLi2 to define its immunological properties. Two mouse mAbs against chCXCLi2 were generated and confirmed to display specific binding with not only recombinants, but endogenous chCXCLi2 by Western blot analysis, ELISA, and immunocytochemistry. Inhibition of chCXCLi2-induced chemotactic activity on peripheral blood lymphocytes, proliferation of chicken macrophage cells and expression of alpha smooth-muscle actin in chicken embryonic fibroblast cells by antibodies indicate that these antibodies are capable of blocking chCXCLi2 bioactivity. These chCXCLi2 mAbs will be useful reagents for future investigations of inflammation in poultry.
白细胞介素-8（IL-8）/CXCL8是一种CXC家族趋化因子，可将淋巴细胞吸引至组织损伤部位，并在炎症反应和伤口愈合中发挥作用。鸡趋化和血管生成因子被称为chCXCLi2，作为人类CXCL8的同源物之一已被研究了20多年。然而，尚未开发出特异性检测chCXCLi2的单克隆抗体（mAb）。在此，我们开发并鉴定了针对chCXCLi2的小鼠mAb，以确定其免疫学特性。通过蛋白质印迹分析、酶联免疫吸附测定和免疫细胞化学，产生了两种针对chCXCLi2的小鼠mAb，并证实它们不仅与重组体特异性结合，还与内源性chCXCLi2特异性结合。抗体对chCXCLi2诱导的外周血淋巴细胞趋化活性、鸡巨噬细胞增殖以及鸡胚成纤维细胞中α平滑肌肌动蛋白表达的抑制作用表明，这些抗体能够阻断chCXCLi2的生物活性。这些chCXCLi2 mAb将成为未来家禽炎症研究的有用试剂。
