Steinbach Falko, Stark Robert, Ibrahim Sherif, Gawad Eman Abd-El, Ludwig Hanns, Walter Jakob, Commandeur Ulrich, Mauel Susanne
Institute for Zoo and Wildlife Research, Alfred-Kowalke-Str. 17, 10315 Berlin, Germany.
Vet Immunol Immunopathol. 2005 Oct 18;108(1-2):227-36. doi: 10.1016/j.vetimm.2005.07.015.
The myeloid cell system comprises of monocytes, macrophages (MPhi), dendritic cells (DC), Kupffer cells, osteoclasts or microglia and is also known as the mononuclear phagocytic system (MPS). Essential cytokines to differentiate or activate these cells include GM-CSF or IL-4. Important markers for characterization include CD1, CD14, CD68, CD163 and CD206. All these markers, however, were not cloned or further characterized in equids by use of monoclonal antibodies earlier. To overcome this problem with the present study, two approaches were used. First, we cloned equine cytokines and markers, and second we analyzed cross-reactivity of human homologues or anti-human monoclonal antibodies. For cloning of equine cytokines and markers, we used degenerate primers delineated from other species, or equine-specific primers based on previous information in Genbank. Flow cytometry was used to determine the expression of markers on myeloid cells. Cross-reactivity could be shown for anti-human CD14, CD163 and mannose receptor (CD206) mAbs. Surface markers such as CD1 and CD68 that distinguish MPhi and DC were cloned and sequenced. According to blast homology, equine CD1a and CD1b could be identified and distinguished. With the resulting information, dendritic cells and macrophages of horses may be characterized.
髓样细胞系统由单核细胞、巨噬细胞（MPhi）、树突状细胞（DC）、库普弗细胞、破骨细胞或小胶质细胞组成，也被称为单核吞噬细胞系统（MPS）。分化或激活这些细胞所需的关键细胞因子包括粒细胞-巨噬细胞集落刺激因子（GM-CSF）或白细胞介素-4（IL-4）。用于细胞鉴定的重要标志物包括CD1、CD14、CD68、CD163和CD206。然而，早期在马属动物中，尚未使用单克隆抗体克隆或进一步鉴定所有这些标志物。为了在本研究中克服这一问题，我们采用了两种方法。第一，我们克隆马属动物的细胞因子和标志物；第二，我们分析人同源物或抗人单克隆抗体的交叉反应性。为了克隆马属动物的细胞因子和标志物，我们使用了从其他物种中设计的简并引物，或基于Genbank中先前信息设计的马属动物特异性引物。采用流式细胞术测定髓样细胞上标志物的表达。抗人CD14、CD163和甘露糖受体（CD206）单克隆抗体可显示交叉反应性。区分MPhi和DC的表面标志物如CD1和CD68被克隆并测序。根据Blast同源性，可鉴定并区分马属动物的CD1a和CD1b。利用所得信息，可对马的树突状细胞和巨噬细胞进行鉴定。
