Quantitation of surface antigens on cultured murine epidermal Langerhans cells: rapid and selective increase in the level of surface MHC products.
Quantitation of surface antigens on cultured murine epidermal Langerhans cells: rapid and selective increase in the level of surface MHC products.
复制标题
培养的鼠表皮朗格汉斯细胞表面抗原的定量：表面 MHC 产物水平的快速和选择性增加。
DOI：
10.1111/1523-1747.ep12456460
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Steinman,RM
中科院分区：
文献类型：
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作者：
Witmer-Pack,MD;Valinsky,J;Olivier,W;Steinman,RM
最近发现，小鼠表皮郎格罕细胞（LC）的功能和表型显着变化时，保持在培养。值得注意的是，辅助细胞功能的主要免疫反应增加，而细胞学标志物，如ATP酶，非特异性酯酶，和Birbeck颗粒丢失。为了进一步分析LC分化，我们使用流式细胞术和一组22种单克隆抗体在单细胞水平上定量表面抗原的变化。一个显著的变化是在培养的第一天Ia抗原（在II类MHC产物上表达）的量增加了5倍。在3 h内增加明显，并在15-24 h达到平台。I-A和I-E产品表现相似。1 μg/ml放线菌酮可阻断Ia的增加。然后通过双色流式细胞术在Ia+LC上监测其他表面抗原的表达。在新鲜分离的LC上检测到低水平的I类（H-2D和H-2K）MHC产物，并且这些抗原在培养的第一天期间也增加了几倍。Fc受体（用2.4G2 mAb鉴定）和F4/80巨噬细胞抗原减少，如先前报告。在新鲜悬浮液中检测到的三种抗原在培养物中以恒定水平表达。这些是C3 bi受体和泛白细胞和交错细胞抗原。一些最初在LC上未发现的白细胞抗原没有出现，包括B220抗B细胞，33 D1抗树突状细胞和CD 4，CD 5，CD 8 T细胞特异性。我们的结论是，培养的LC的表面进行选择性的变化，在文化。结果，细胞富含Ia和H-2，并具有可检测的C3 bi受体，但具有很少或没有LFA-1、Ti、CD 4、5和8、33 D1、2.4G2、F4/80和B220抗原。
It was recently discovered that murine epidermal Langerhans cells (LC) changed significantly in function and phenotype when maintained in culture. Notably, accessory cell function for primary immune responses increased while cytologic markers like ATPase, nonspecific esterase, and Birbeck granules were lost. To further analyze LC differentiation, we used flow cytometry and a panel of 22 monoclonal antibodies to quantitate changes in surface antigens at the single-cell level. A striking change was a fivefold increase in the amount of Ia antigens (which are expressed on class II MHC products) during the first day of culture. The increase was evident within 3 h and reached a plateau at 15-24 h. Both I-A and I-E products behaved similarly. The increase in Ia was blocked by 1 μg/ml cycloheximide. Expression of other surface antigens was then monitored on Ia+LC by two-color flow cytometry. Low levels of class I (H-2D and H-2K) MHC products were detected on freshly isolated LC, and these antigens also increased severalfold during the first day of culture. Fc receptors (identified with the 2.4G2 mAb) and the F4/80 macrophage antigen decreased, as reported previously. Three antigens that were detected in fresh suspensions were expressed at constant levels in culture. These were the C3bi receptor and the pan leukocyte and interdigitating cell antigens. Several leukocyte antigens that were not found initially on LCs did not appear, including B220 anti-B cell, 33D1 anti-dendritic cell, and CD4, CD5, CD8 T-cell specificities. We conclude that the surface of cultured LCs undergoes selective changes in culture. As a result, the cells are rich in Ia and H-2 and have detectable C3bi receptors, but have little or no LFA-1, Ti, CD 4, 5, and 8, 33D1, 2.4G2, F4/80, and B220 antigens.