Flow cytometric analysis of epidermal subpopulations from normal and psoriatic skin using monoclonal antibodies against intermediate filaments.
Keratin-type intermediate filament proteins show characteristic expression in normal and pathologic epidermis. Some keratins are restricted to the basal cell layers, and others occur exclusively in the suprabasal compartment. SDS-gel-electrophoresis and immunohistochemistry are generally used for the assessment of keratin profiles and their localizations. In the present investigation, flow cytometric analysis of four different monoclonal antibodies (MAb) against intermediate filament-type proteins, in addition to measurement of relative DNA content, was performed on cell suspensions derived from lesional and clinically uninvolved skin of psoriatic patients and from skin of healthy controls. MAb Ks8.12, reacting with keratins 13 and 16, was used as a marker for hyperproliferation. Pab601 recognizes the basal cell layer(s) of human epidermis. Keratin 10 expression as a marker of keratinization was quantified with RKSE60 and the anti-vimentin MAb MVI was used as a marker for non-keratinocytes. Psoriatic skin showed significantly reduced numbers of RKSE60-positive cells and MVI-positive cells compared with normal skin. In contrast to normal skin and uninvolved skin of psoriatic patients in which only a minority of the cells were Ks8.12 positive, up to 60% of the cell population in psoriatic lesions bound with MAb. Simultaneous measurement of relative DNA content and MAb binding showed that Pab601 binding was associated with cells in S-phase and G2M-phase of the cell cycle, whereas RKSE60 and Ks8.12 binding were associated with diploid cells. Multiparameter flow cytometry allows quantitative population analysis that could lead to a better understanding of the complex mechanisms of epidermal growth control under normal and pathologic conditions.
van Erp
P E
PE
Department of Dermatology, University Hospital, Nijmegen, The Netherlands.
Rijzewijk
J J
JJ
Boezeman
J B
JB
Leenders
J
J
de Mare
S
S
Schalkwijk
J
J
van de Kerkhof
P C
PC
Ramaekers
F C
FC
Bauer
F W
FW
D000328:Adult; D000911:Antibodies, Monoclonal; D004247:DNA / Q000032:analysis; D004817:Epidermis / Q000032:analysis* / Q000473:pathology; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D007381:Intermediate Filament Proteins / Q000032:analysis*; D007399:Interphase; D007633:Keratins / Q000032:analysis; D008875:Middle Aged; D011565:Psoriasis / Q000378:metabolism* / Q000473:pathology
D000911:Antibodies, Monoclonal; D007381:Intermediate Filament Proteins; D007633:Keratins; D004247:DNA