Mac-2, a novel 32,000 Mr mouse macrophage subpopulation-specific antigen defined by monoclonal antibodies.
Mac-2, a novel 32,000 Mr mouse macrophage subpopulation-specific antigen defined by monoclonal antibodies.
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DOI：
10.4049/jimmunol.128.3.1221
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发表时间：
1982-03
影响因子：
4.4
通讯作者：
M. Ho;T. Springer
中科院分区：
文献类型：
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作者：
M. Ho;T. Springer
通过将小鼠骨髓瘤细胞与免疫吸附纯化的巨噬细胞糖蛋白免疫的大鼠脾细胞融合，获得两种单克隆抗体 M3/31 和 M3/38。共沉淀实验表明，这两种抗体识别的抗原决定簇位于相同的分子种类上，称为 Mac-2，Mac-2 是一种 32,000 Mr 的抗原，由硫代乙醇酸诱导的巨噬细胞合成并在其表面表达，如 [35S]-蛋氨酸和 125I 标记所示。饱和结合实验表明，巯基乙酸引发的巨噬细胞表达 1.7 X 10(5) Mac-2 位点/细胞。硫代乙醇酸盐引发的巨噬细胞对 125I 标记的 M3/38 MAb 有很强的吸收能力。肾脏也有吸收能力；然而，有证据表明这种吸收的非特异性。淋巴结和胸腺呈阴性，而脾脏和骨髓的吸收能力较弱，可能是由于基质细胞的原因。非淋巴组织，如肺、肝、心脏和脑，表现出轻微或没有吸收能力。通过免疫荧光流式细胞术检测，来自脾脏、骨髓、胸腺和外周淋巴结的细胞悬液中 Mac-2- 含量大于 99%。相比之下，巯基乙酸引发的巨噬细胞中 Mac-2 强阳性率超过 96%。只有 20% 的蛋白胨引发的细胞呈弱阳性，而常驻腹膜巨噬细胞和单核细胞增生李斯特菌、Con A 或 LPS 引发的其他巨噬细胞的阴性率大于 98%。 [35S]-蛋氨酸标记的 Mac-2 的 SDS-PAGE 显示，硫代乙醇酸诱导的巨噬细胞合成的 Mac-2 比其他腹膜巨噬细胞亚群多 10 至 30 倍，而所有类型的腹膜巨噬细胞在其表面合成和表达相似量的 Mac-1 抗原。因此，巨噬细胞中的 Mac-2 抗原仅响应特定的分化信号而被诱导。
Two monoclonal antibodies, M3/31 and M3/38, were obtained by fusion of mouse myeloma cells with rat spleen cells immunized to immunoadsorbent-purified macrophage glycoproteins. Co-precipitation experiments show that antigenic determinants recognized by these two antibodies reside on the same molecular species, termed Mac-2, Mac-2, an antigen of 32,000 Mr, is synthesized by and expressed on the surface of thioglycollate-elicited macrophages as shown by [35S]-methionine and 125I labeling. Saturation binding experiments show that thioglycollate-elicited macrophages express 1.7 X 10(5) Mac-2 sites/cell. Thioglycollate-elicited macrophages are strongly absorptive for 125I-labeled M3/38 MAb. Kidneys are also absorptive; however, evidence is presented pointing to the nonspecificity of this absorption. Lymph node and thymus are negative, whereas spleen and bone marrow are weakly absorptive, probably due to stromal cells. Nonlymphoid tissues, such as lung, liver, heart, and brain, exhibit slight or no absorbing capacity. Cell suspensions from spleen, bone marrow, thymus, and peripheral lymph node are greater than 99% Mac-2- by immunofluorescent flow cytometry. In contrast, thioglycollate-elicited macrophages are greater than 96% strongly positive for Mac-2. Only 20% of peptone-elicited cells are weakly positive, whereas resident peritoneal macrophages and other macrophage elicited by Listeria monocytogenes, Con A, or LPS are greater than 98% negative. SDS-PAGE of [35S]-methionine-labeled Mac-2 shows that thioglycollate-elicited macrophages synthesize 10- to 30-fold more Mac-2 than other peritoneal macrophage subpopulations, whereas all types of peritoneal macrophages synthesize and express on their surfaces similar amounts of the Mac-1 antigen. Mac-2 antigen is therefore induced in macrophages only in response to specific differentiative signals.