Monitoring hyperproliferative disorders in human skin: flow cytometry of changing cytokeratin expression.

Abstract

Monitoring dynamics of different cell populations in solid tissues using flow cytometry has several limitations. The interaction and changes in epidermal subpopulations in hyperproliferative skin disorders such as psoriasis, a very common chronic inflammatory skin disease, may, however, elucidate the role of different cell populations in the pathogenesis and the effect of therapy in these disorders. We describe a new, simple method for identifying and quantifying different epidermal subpopulations in hyperproliferative skin disorders and an in vivo model for epidermal hyperproliferation by using six-parameter assessment and three-color flow cytometry. Epidermal single-cell suspensions were derived from normal human skin before and after standardized injury and from untreated and treated psoriatic lesions. Samples were stained with anti-cytokeratins 6 (K6) and 10 (K10), anti-vimentin, and 4',6-diamidino-2-phenylindole. With three-color flow cytometry, different epidermal subpopulations were further defined by six different parameters. Correlations were found for mild psoriasis and K10(+)K6(-) cells and for moderate psoriasis and K10(-)K6(+) cells. Treatment of mild psoriasis resulted in a 70% increase of the K10(+)K6(-) cells and an 18% decrease of the K10(-)K6(-) cells, whereas treatment of more severe psoriasis resulted in a 77% increase of the K10(+)K6(-) cells and a 33% decrease of the K10(-)K6(+) cells. The tape-stripping model showed changes in suprabasal keratin expression before proliferative activity. The present flow cytometric assay permits simultaneous quantification of epidermal differentiation, inflammation, proliferation, and hyperproliferation-associated keratinization and provides information on pathogenesis and therapy of hyperproliferative skin disorders.