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OKT6 is not superior to HLA‐DR or ATPase as a marker for Langerhans' cells in normal human epidermis
Author(s) -
LIU H.H.,
SCHROETER A.L.,
MULLER S.A.
Publication year - 1986
Publication title -
clinical and experimental dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.587
H-Index - 78
eISSN - 1365-2230
pISSN - 0307-6938
DOI - 10.1111/j.1365-2230.1986.tb00453.x
Subject(s) - epidermis (zoology) , langerhans cell , staining , monoclonal antibody , fluorescein isothiocyanate , antigen , hla dr , biology , antibody , frozen section procedure , microbiology and biotechnology , pathology , chemistry , immunology , medicine , anatomy , fluorescence , physics , quantum mechanics
Summary Previous studies have suggested that HLA‐DR + /OKT6 + (DR + T6 + ) and DR − T6 + subsets of Langerhans' cells exist in normal epidermis. Using parallel double‐staining techniques on epidermal sheets as well as on frozen sections as a control, we have been able to demonstrate that 97–100% of T6 + dendritic cells are DR + . This finding suggests that DR T6 + and DR − T6 + subsets of Langerhans' cells do not exist. In contrast to T6 monoclonal antibody (mAb) directly labelled with fluorescein isothiocyanate, DR mAb indirectly labelled with rhodamine usually stained Langerhans' cells unevenly in the human epidermal sheets (because Langerhans' cells localize throughout the epidermis); however, this effect became much less conspicuous on 4‐μm frozen vertical sections and on thinner epidermal sheets. DR antigen density appears to be similar among Langerhans' cells. The even staining with T6 antibody suggests that ii may have a higher affinity for the T6 antigen than DR antibody has for the DR antigen. In normal skin, no significant difference was found among the three different surface markers of Langerhans' cells. ATPase staining on the epidermal sheet, already used for the double staining, significantly underestimated the presence of Langerhans' cells. Previous studies (Muhlbauer et al. , 1982; Harrist et al. , 1983) have suggested that not all epidermal Langerhans' cells and indeterminate cells are HLA‐DR + and that HLA‐DR + OKT6 + (DR + T6 + and DR − T6 + subsets exist. With a four‐step peroxidase‐antiperoxidase method using either anti‐T6 or anti‐DR on separate frozen sections, the ratio of DR + to T6 + Langerhans' cells varied from 16% to 60% (Muhlbauer et al. , 1982; Harrist et al. , 1983). In contrast, by the method of double labelling of epidermal cells with DR and T6, Fithian and colleagues (1981) demonstrated that these two antibodies stained identical cells in tissue section. Vertically sectioned Epon‐embedded specimens, however, may not be suitable for the enumeration of Langerhans' cells in skin specimens (Horton, Allen & MacDonald, 1983). Recently, Dezutter‐Dambuyant et al , (1984b) reported that about 97% of the labelled dendritic epidermal cells in dispersed human epidermal cell suspensions were DR + T6 + , but Berman and colleagues (1985) found that only approximately 50% of T6 + Langerhans' cells were This discrepancy may be due to the fact that, in the absence of dermis, Langerhans' cells rapidly lose the ability to produce the DR antigen (Czernielewski, Demarchez & Prunieras, 1984; Hefton et al. , 1984), and the cell membrane of Langerhans' cells could be damaged by the disaggregating effect of trypsin. Because of these disadvantages, we used parallel double‐labelling experiments to enumerate Langerhans' cells on the epidermal sheet. To compare with the above two surface markers of Langerhans' cells, a modified ATPase staining technique was used on the sheet with preceding double stains or on a separate epidermal sheet.