Recent advances of ultrastructural immunocytochemistry of epidermal Langerhans cells

Using electron microscopy, the immunological visualization of the membrane antigens of Langerhans cells (LC) can be performed by immunoperoxidase and immunogold techniques. The immunoperoxidase labelling permits the identification of only one antigen and the observation of qualitative variations of surface antigens. The immunogold method allows the identification of one antigen or simultaneously two or three surface antigens using gold particles of various sizes. This technique can be used to quantify the surface density of antigens on the cell membrane. The simultaneous identification of different surface antigens can be correlated with the ultrastructural characteristics of the cells. Using this technique we have recently demonstrated the existence of LC subsets in normal epidermis, and the presence of circulating T6-positive cells in normal subjects. In addition, a very low density of T4 antigenic sites on the LC membrane surface was observed. Several problems of a double-labelling immunogold technique related to steric hindrance and current artifacts are discussed.