Witmer-Pack M D, Valinsky J, Olivier W, Steinman R M
Laboratory of Cellular Physiology and Immunology, Rockefeller University, New York, New York.
J Invest Dermatol. 1988 Mar;90(3):387-94. doi: 10.1111/1523-1747.ep12456460.
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 microgram/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, CD4, 5, and 8, 33D1, 2.4G2, F4/80, and B220 antigens.
最近发现，小鼠表皮朗格汉斯细胞（LC）在培养过程中功能和表型发生了显著变化。值得注意的是，初级免疫反应的辅助细胞功能增强，而诸如ATP酶、非特异性酯酶和伯贝克颗粒等细胞学标志物则消失。为了进一步分析LC的分化，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，培养第一天Ia抗原（在II类MHC产物上表达）的量增加了五倍。这种增加在3小时内就很明显，并在15 - 24小时达到平台期。I - A和I - E产物表现相似。1微克/毫升的环己酰亚胺可阻断Ia的增加。然后通过双色流式细胞术监测Ia + LC上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，这些抗原在培养第一天也增加了几倍。如先前报道的那样，Fc受体（用2.4G2单克隆抗体鉴定）和F4/80巨噬细胞抗原减少。在新鲜悬浮液中检测到的三种抗原在培养中表达水平恒定。这些是C3bi受体以及全白细胞和交错突细胞抗原。最初在LC上未发现的几种白细胞抗原也未出现，包括抗B细胞的B220、抗树突状细胞的33D1以及CD4、CD5、CD8 T细胞特异性抗原。我们得出结论，培养的LC表面在培养过程中发生了选择性变化。结果，这些细胞富含Ia和H - 2，具有可检测的C3bi受体，但几乎没有或没有LFA - 1、Ti、CD4、5和8、33D1、2.4G2、F4/80和B220抗原。
