title: Enhanced binding of peripheral blood mononuclear leukocytes to gamma-interferon-treated cultured keratinocytes.
abstract: Dermatopathologists observe mononuclear leukocytes in close apposition to keratinocytes (KCs) in graft versus host disease and in other lymphocyte-mediated skin diseases, such as lichen planus, erythema multiforme, and lupus erythematosus. Since the KCs are Class II histocompatibility antigen (HLA-DR) positive in these diseases (indicating local production of gamma interferon, IFN-gamma, by activated T-cells), we sought to determine whether IFN-gamma treatment of KCs would influence the ability of allogeneic peripheral blood mononuclear leukocytes (PBMLs) to adhere to cultured KCs in vitro. The adherence of PBMLs to KC monolayers was determined by the three following methods: (a) methanol fixation of the washed KCs (after PBML incubation), followed by hematoxylin-eosin staining and direct counting of adherent PBMLs; (b) fluorescein isothiocyanate (FITC) labeling of PBML, followed by measuring the amount of FITC-PBML bound to KCs after washing either by direct visualization with a fluorescence microscope; or by (c) quantitative fluorescence spectroscopy following lysis of the adherent cells. While untreated KCs bound allogeneic PBMLs minimally 15-120 min at 37 degrees C, pretreatment of the KCs with IFN-gamma (300 U/ml, 3 days) produced significantly increased binding of the PBMLs by approximately fivefold. By contrast, IFN-alpha and IFN-beta (10(3) U/ml) had no effect. Also, despite the induction of HLA-DR on cultured human fibroblasts, no increased binding of PBMLs after IFN-gamma treatment was observed. The selective ability of IFN-gamma to produce a marked increase in adherence between KCs and PBMLs suggests a new role for IFN-gamma in the immunobiology of the skin.
mesh_terms: D002448:Cell Adhesion; D002478:Cells, Cultured; D000078404:Epidermal Cells; D005455:Fluorescent Antibody Technique; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology*; D007633:Keratins / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000276:immunology*; D008856:Microscopy, Fluorescence; D011994:Recombinant Proteins; D012867:Skin / Q000276:immunology
chemical_list: D011994:Recombinant Proteins; D007633:Keratins; D007371:Interferon-gamma