Morris J, Alaibac M, Jia M H, Chu T
Unit of Dermatology, Royal Postgraduate Medical School, London, England.
J Invest Dermatol. 1992 Aug;99(2):237-40. doi: 10.1111/1523-1747.ep12650459.
Our knowledge of the functional activity of the epidermal Langerhans cell has been severely hampered by the lack of an easy method of purification of these cells that is both efficient and reproducible. In the present study we have used immunomagnetic beads directly conjugated to an IgM class mouse anti-human human leukocyte antigen DR monoclonal antibody to positively select human Langerhans cells from an epidermal cell suspension. Cells were then treated with a high-affinity polyclonal anti-mouse immunoglobulin that detached the beads by competing with the antigen for the antigen-binding site on the monoclonal antibody. This procedure allowed removal of the immunomagnetic beads, leaving Langerhans cells free from bound antibody. Recovery of Langerhans cells ranged from 40 to 80% of the starting number of Langerhans cells. The resulting cells were up to 99% CD1a positive and showed potent functional activity in the allogeneic mixed epidermal cell - lymphocyte reaction. Keratinocytes were shown to exert a profound inhibitory effect on Langerhans cell function that could not be prevented by indomethacin. This method is technically simple and allows good recovery of a highly purified population of Langerhans cells that are functionally active.
由于缺乏一种简便、高效且可重复的纯化表皮朗格汉斯细胞的方法，我们对其功能活性的了解受到了严重阻碍。在本研究中，我们使用直接偶联有IgM类小鼠抗人白细胞抗原DR单克隆抗体的免疫磁珠，从表皮细胞悬液中阳性选择人朗格汉斯细胞。然后用高亲和力的多克隆抗小鼠免疫球蛋白处理细胞，该免疫球蛋白通过与抗原竞争单克隆抗体上的抗原结合位点来使磁珠脱落。此过程可去除免疫磁珠，使朗格汉斯细胞不带有结合抗体。朗格汉斯细胞的回收率为起始朗格汉斯细胞数量的40%至80%。所得细胞CD1a阳性率高达99%，并在同种异体混合表皮细胞 - 淋巴细胞反应中表现出强大的功能活性。角质形成细胞对朗格汉斯细胞功能具有显著的抑制作用，吲哚美辛无法阻止这种作用。该方法技术简单，能够很好地回收功能活跃的高度纯化的朗格汉斯细胞群体。
