Garcia-Morales Carla, Rothwell Lisa, Moffat Lindsey, Garceau Valerie, Balic Adam, Sang Helen M, Kaiser Pete, Hume David A
The Roslin Institute & Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian EH25 9RG, UK.
Dev Comp Immunol. 2014 Feb;42(2):278-85. doi: 10.1016/j.dci.2013.09.011. Epub 2013 Sep 29.
Macrophages contribute to innate and acquired immunity as well as many aspects of homeostasis and development. Studies of macrophage biology and function in birds have been hampered by a lack of definitive cell surface markers. As in mammals, avian macrophages proliferate and differentiate in response to CSF1 and IL34, acting through the shared receptor, CSF1R. CSF1R mRNA expression in the chicken is restricted to macrophages and their progenitors. To expedite studies of avian macrophage biology, we produced an avian CSF1R-Fc chimeric protein and generated a monoclonal antibody (designated ROS-AV170) against the chicken CSF1R using the chimeric protein as immunogen. Specific binding of ROS-AV170 to CSF1R was confirmed by FACS, ELISA and immunohistochemistry on tissue sections. CSF1 down-regulated cell surface expression of the CSF1R detected with ROS-AV170, but the antibody did not block CSF1 signalling. Expression of CSF1R was detected on the surface of bone marrow progenitors only after culture in the absence of CSF1, and was induced during macrophage differentiation. Constitutive surface expression of CSF1R distinguished monocytes from other myeloid cells, including heterophils and thrombocytes. This antibody will therefore be of considerable utility for the study of chicken macrophage biology.
巨噬细胞在天然免疫和获得性免疫以及体内平衡和发育的许多方面都发挥着作用。由于缺乏明确的细胞表面标志物,鸟类巨噬细胞生物学和功能的研究受到了阻碍。与哺乳动物一样,禽类巨噬细胞通过共同受体CSF1R对CSF1和IL34作出反应而增殖和分化。鸡体内CSF1R mRNA的表达仅限于巨噬细胞及其祖细胞。为了加快禽类巨噬细胞生物学的研究,我们制备了一种禽类CSF1R-Fc嵌合蛋白,并使用该嵌合蛋白作为免疫原产生了一种针对鸡CSF1R的单克隆抗体(命名为ROS-AV170)。通过流式细胞术、酶联免疫吸附测定和组织切片免疫组织化学证实了ROS-AV170与CSF1R的特异性结合。CSF1下调了用ROS-AV170检测到的CSF1R的细胞表面表达,但该抗体并未阻断CSF1信号传导。仅在无CSF1的培养基中培养后,骨髓祖细胞表面才检测到CSF1R的表达,且在巨噬细胞分化过程中诱导表达。CSF1R的组成型表面表达可将单核细胞与其他髓细胞区分开来,包括异嗜性粒细胞和血小板。因此,该抗体在鸡巨噬细胞生物学研究中将具有相当大的用途。