Koch A E, Burrows J C, Domer P H, Ashmun R A, Look A T, Leibovich S J
Northwestern University Medical School, Department of Medicine, Chicago, IL 60611.
Pathobiology. 1992;60(2):59-67. doi: 10.1159/000163699.
We have selected several monoclonal antibodies (mAbs) producing using human rheumatoid arthritis (RA) synovial macrophages (m phi s) as immunogen. Of these, mAbs 8H2, 10G7 and 10G9 showed cross reactivity with endothelium, suggesting common antigens between these cell types. We have determined the spectrum of reactivity of these mAbs on hematopoietic cell lines, peripheral blood cells, and inflammatory and non-inflammatory tissues by immunohistochemistry. MAb 8H2 does not react with the myeloid cell lines HL60 (myelocytic), U937 (histiocytic lymphoma), and K562 (erythroleukemia), or with peripheral blood cells. In normal and inflamed tissue sections, mAb 8H2 reacts with m phi s and endothelial cells. In contrast, mAb 10G7 does not react with peripheral blood cells, but reacts with HL60, U937, and K562 cell lines, as well as with m phi s and endothelial cells in inflamed and noninflamed tissues. MAb 10G9 does not react with myeloid cell lines, but reacts with monocytes and platelets in peripheral blood. In both normal and inflamed tissues, mAb 10G9 reacts with m phi s and endothelial cells. The antigens identified by these three mAbs were characterized biochemically, by enzymatic digestion of RA synovial tissue m phi s followed by a cellular ELISA, as well as by reactivity of the mAbs with NIH-3T3 cells genetically engineered to express known myeloid antigens. These mAbs reacted with protein or glycoprotein antigens distinct from the known myeloid antigens CD13, CD14, CD33, CD34, CD36, and c-fms. These mAbs should prove to be a valuable tool for studying m phi s and endothelial cells and their shared antigenic determinants.
我们选用了几种以人类类风湿性关节炎(RA)滑膜巨噬细胞(m phi s)作为免疫原制备的单克隆抗体(mAbs)。其中,单克隆抗体8H2、10G7和10G9与内皮细胞表现出交叉反应,提示这些细胞类型之间存在共同抗原。我们通过免疫组织化学确定了这些单克隆抗体在造血细胞系、外周血细胞以及炎性和非炎性组织上的反应谱。单克隆抗体8H2不与髓系细胞系HL60(髓细胞性)、U937(组织细胞淋巴瘤)和K562(红白血病)反应,也不与外周血细胞反应。在正常和炎症组织切片中,单克隆抗体8H2与m phi s和内皮细胞反应。相比之下,单克隆抗体10G7不与外周血细胞反应,但与HL60、U937和K562细胞系反应,也与炎症和非炎症组织中的m phi s和内皮细胞反应。单克隆抗体10G9不与髓系细胞系反应,但与外周血中的单核细胞和血小板反应。在正常和炎症组织中,单克隆抗体10G9都与m phi s和内皮细胞反应。通过对RA滑膜组织m phi s进行酶消化后进行细胞ELISA,以及通过这些单克隆抗体与经基因工程改造以表达已知髓系抗原的NIH-3T3细胞的反应性,对这三种单克隆抗体识别的抗原进行了生化特征分析。这些单克隆抗体与不同于已知髓系抗原CD13、CD14、CD33、CD34、CD36和c-fms的蛋白质或糖蛋白抗原反应。这些单克隆抗体将被证明是研究m phi s和内皮细胞及其共同抗原决定簇的有价值工具。