A monoclonal antibody (3G5)-defined ganglioside antigen is expressed on the cell surface of microvascular pericytes.
A monoclonal antibody (3G5)-defined ganglioside antigen is expressed on the cell surface of microvascular pericytes.
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DOI：
10.1084/jem.167.3.1003
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发表时间：
1988-03-01
期刊：
影响因子：
--
通讯作者：
King GL
中科院分区：
文献类型：
--
作者：
Nayak RC;Berman AB;George KL;Eisenbarth GS;King GL
微血管周细胞的体外鉴定主要依赖于形态学特征和生长动力学，因为这些细胞的免疫化学标记物很少。因此，尝试鉴定有助于快速鉴定和富集血管细胞培养物中视网膜毛细血管周细胞的mAb试剂。用牛视网膜毛细血管周细胞的分离培养物筛选通过异种免疫接种具有各种组织的小鼠而产生的一组mAb的免疫反应性。通过间接免疫荧光观察到来自该组的两种抗体（3G 5和HISL-8）与周细胞反应。选择mAb 3G 5用于进一步研究。mAb 3G 5不与平滑肌细胞、内皮细胞或视网膜色素内皮细胞的解离培养物反应。周细胞3G 5抗原对胰蛋白酶的作用不敏感;因此，mAb 3G 5用于通过流式细胞术从胰蛋白酶化的混合视网膜细胞培养物中选择性地纯化周细胞。3G 5+周细胞（代表混合视网膜细胞培养物中8%的细胞）富集至少9倍以代表大于70%的细胞。mAb 3G 5在体内染色视网膜毛细血管，其荧光分布与周细胞染色一致。发现培养的周细胞的3G 5抗原是神经节苷脂标记物GM 1和GM 2之间的迁移率中间的糖脂。
The identification of microvascular pericytes in vitro relies principally on morphological characteristics and growth dynamics, as there is a paucity of immunochemical markers for these cells. Consequently, an attempt was made to identify mAb reagents that would aid in both the rapid identification and enrichment of retinal capillary pericytes in vascular cell cultures. A panel of mAbs raised by xenogeneic immunization of mice with various tissues was screened for immunoreactivity with dissociated cultures of bovine retinal capillary pericytes. Two antibodies from the panel (3G5 and HISL-8) were seen to react with pericytes by indirect immunofluorescence. The mAb 3G5 was selected for further study. mAb 3G5 did not react with dissociated cultures of smooth muscle cells, endothelial cells, or retinal pigmented endothelial cells. The pericyte 3G5 antigen was insensitive to the action of trypsin; therefore, mAb 3G5 was used to selectively purify pericytes from trypsinized mixed retinal cell cultures by flow cytometry. 3G5+ pericytes (representing 8% of cells in a mixed retinal cell culture) were enriched at least nine-fold to represent greater than 70% of cells. The mAb 3G5 stained retinal capillaries in vivo with a fluorescence distribution consistent with pericyte staining. The 3G5 antigen of cultured pericytes was found to be a glycolipid of mobility intermediate between ganglioside markers GM1 and GM2.