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FITC-insulin binding to normal and psoriatic epidermis
Acta histochem. 85, 223-226 (1989) VEB Gustav Fischer Verlag Jena
C1~nic
and Policlinic of Dermatology, Department of Medicine, the Friedrich Schiller University, Jena, GDR
FITC-insulin binding to normal and psoriatic epidermis I) By UWE WOLLlNA, CHRISTINA HIPLER and BURKHARD KNOPF With one Figure (Received July 4, 1988)
Summary FITC-insulin was prepared and applied to frozen human skin sections. In normal and nonlesional psoriatic epidermis, the binding was cytoplasmic in suprabasal cells. In psoriatic lesions, fairly stained or even unstained cells were littered in suprabasal epidermis indicating the expansion of selected basal cell qualities into the stratum spinosum.
Introduction The binding of FITC-insulin has been related to the expression of insulin receptors and the degree of cellular differentiation (FENG et al. 1987; VERRANDO and ORTONNE 1985). Recently, BELLOMO et al. (1982) reported about a numerical loss and a decrease of receptor affinity in fibroblasts from psoriatic skin. Thus, we employed the staining technique with FITC-insulin to get information about epidermal cell differentiation in psoriasis.
Material and methods Skin biopsies from lesional and nonlesional psoriatic skin and healthy volunteers were involved. The tissues were snap frozen in liquid nitrogen and processed to 4 !-lm thick unfixed frozen sections. Monocomponent insulin obtained from NOVO. Mainz (FRG). was mixed with FITC (33 mg insulin + 12 mg FITC) and stirred at 3°C for 24 h. The mixture was purified on a Bio-gel P-2 column with PBS. pH = 7.2, for eluation. The solution was dialyzed against glycol 20,000 in PBS. Protein (P) concentration = 0.5Ilg/ml, fluorescein/P ratio = 0.22 x 10-\ working dilution = I: 10. Sections were incubated for 30 min at room temperature. In parallel. inhibition experiments with unlabelled Binsulin (VEB Berlin-Chemie. GDR) were performed.
Results F1TC-insulin was fixed by all samples in a cytoplasmic manner. Normal and nonlesional psoriatic epidermis disclosed a suprabasal staining (Fig. I a, b). Psoriatic lesions showed littered suprabasal cells with fainty or even no staining. The overall staining intensity was decreased, which was most pronounced in the rete ridges (Fig. I c). Inhibition experiments prevented epidermal staining completely.
I) Supported by GDR research grant M51.
224
U. WOLLINA. CHR. HIPLER and B. KNOPF
Fig. I. Human epidennis stained with FITC-insulin. a Nonnal epidermis. b Nonlesional psoriatic epidennis. cRete ridge of a psoriatic lesion.
FITC-insulin binding
225
Discussion The binding ofFlTC-insulin has been related to keratinocyte's differentiation (FENG et al. 1987). We observed a disturbed epidermal staining in psoriatic lesions versus controls: Littered suprabasal cells disclosed a fainty or even absent fluorescence, whereas in controls the entire suprabasal epidermis disclosed a strong cytoplasmic labelling (Fig. I). Thus, we gained further evidence for an altered epidermal differentiation in lesional psoriatic epidermis accompanied by preservation of selected basal cell qualities in the stratum spinosum (HEENEN and GALAND 1984; TANK ct al. 19R6).
References BELLOMO, G., FRATINO, P., LOCURTO, F., PELFINI, c., VIGUINt, M., and SERRI, F., Insulin receptors in psoriasis. In: Psoriasis. Proceedings of the 3rd International Symposium (Eds.: FARBER, E. M., and Cox, A. J.). Grune & Stratton, New York 1982, pp. 273-274. FENG. H.- W., JIN, X.-P., and BERNSTEIN, I. A., Relationship between cell differentiation and binding of fluorescein isothiocyanate (FITC)-conjugated insulin of keratinocytes in culture. J. Invest. Dermatol 89, 73-77 (1987). HEE EN. M., and G.~LAND. P., On cell kinetics in psoriasis. Brit. J. Dermatol. 110,241-245 (1984).
226
U. WOLLlNA, CHR. HIPLER and B. KNOPF
TANK, B., VAN DUK, G., VAN DER SLUIS, J. J., VAN DER KAMP, A. W. M., SCHONK, D., VERSNEL, M., STOLZ, E., and VAN JOOST, T., The detection of basal cell determinants in human psoriatic skin: a monoclonal antibody study. Arch. Dermatol. Res. 279,68-70 (1986). VERRANDO, P., and ORTONNE, J. P., Insulin binding properties of normal and transformed human epidermal cultured keratinocytes. J. Invest. Dermatol. 85, 328- 332 (1985). Authors' address: Dr. sc. med. U. WOLLlNA, Universitats-Hautklinik, Erfurter StraBe 35, Jena, DDR - 6900.
Acta histochem. 1'01.
HAHX
,"ox
Piette V
8;j
])OBSCHE, F.~LT,
an,l
ZtHLKE
VEll II tSTA v FISCHElt VEltLAG .TEXA
C1~nic
and Policlinic of Dermatology, Department of Medicine, the Friedrich Schiller University, Jena, GDR
FITC-insulin binding to normal and psoriatic epidermis I) By UWE WOLLlNA, CHRISTINA HIPLER and BURKHARD KNOPF With one Figure (Received July 4, 1988)
Summary FITC-insulin was prepared and applied to frozen human skin sections. In normal and nonlesional psoriatic epidermis, the binding was cytoplasmic in suprabasal cells. In psoriatic lesions, fairly stained or even unstained cells were littered in suprabasal epidermis indicating the expansion of selected basal cell qualities into the stratum spinosum.
Introduction The binding of FITC-insulin has been related to the expression of insulin receptors and the degree of cellular differentiation (FENG et al. 1987; VERRANDO and ORTONNE 1985). Recently, BELLOMO et al. (1982) reported about a numerical loss and a decrease of receptor affinity in fibroblasts from psoriatic skin. Thus, we employed the staining technique with FITC-insulin to get information about epidermal cell differentiation in psoriasis.
Material and methods Skin biopsies from lesional and nonlesional psoriatic skin and healthy volunteers were involved. The tissues were snap frozen in liquid nitrogen and processed to 4 !-lm thick unfixed frozen sections. Monocomponent insulin obtained from NOVO. Mainz (FRG). was mixed with FITC (33 mg insulin + 12 mg FITC) and stirred at 3°C for 24 h. The mixture was purified on a Bio-gel P-2 column with PBS. pH = 7.2, for eluation. The solution was dialyzed against glycol 20,000 in PBS. Protein (P) concentration = 0.5Ilg/ml, fluorescein/P ratio = 0.22 x 10-\ working dilution = I: 10. Sections were incubated for 30 min at room temperature. In parallel. inhibition experiments with unlabelled Binsulin (VEB Berlin-Chemie. GDR) were performed.
Results F1TC-insulin was fixed by all samples in a cytoplasmic manner. Normal and nonlesional psoriatic epidermis disclosed a suprabasal staining (Fig. I a, b). Psoriatic lesions showed littered suprabasal cells with fainty or even no staining. The overall staining intensity was decreased, which was most pronounced in the rete ridges (Fig. I c). Inhibition experiments prevented epidermal staining completely.
I) Supported by GDR research grant M51.
224
U. WOLLINA. CHR. HIPLER and B. KNOPF
Fig. I. Human epidennis stained with FITC-insulin. a Nonnal epidermis. b Nonlesional psoriatic epidennis. cRete ridge of a psoriatic lesion.
FITC-insulin binding
225
Discussion The binding ofFlTC-insulin has been related to keratinocyte's differentiation (FENG et al. 1987). We observed a disturbed epidermal staining in psoriatic lesions versus controls: Littered suprabasal cells disclosed a fainty or even absent fluorescence, whereas in controls the entire suprabasal epidermis disclosed a strong cytoplasmic labelling (Fig. I). Thus, we gained further evidence for an altered epidermal differentiation in lesional psoriatic epidermis accompanied by preservation of selected basal cell qualities in the stratum spinosum (HEENEN and GALAND 1984; TANK ct al. 19R6).
References BELLOMO, G., FRATINO, P., LOCURTO, F., PELFINI, c., VIGUINt, M., and SERRI, F., Insulin receptors in psoriasis. In: Psoriasis. Proceedings of the 3rd International Symposium (Eds.: FARBER, E. M., and Cox, A. J.). Grune & Stratton, New York 1982, pp. 273-274. FENG. H.- W., JIN, X.-P., and BERNSTEIN, I. A., Relationship between cell differentiation and binding of fluorescein isothiocyanate (FITC)-conjugated insulin of keratinocytes in culture. J. Invest. Dermatol 89, 73-77 (1987). HEE EN. M., and G.~LAND. P., On cell kinetics in psoriasis. Brit. J. Dermatol. 110,241-245 (1984).
226
U. WOLLlNA, CHR. HIPLER and B. KNOPF
TANK, B., VAN DUK, G., VAN DER SLUIS, J. J., VAN DER KAMP, A. W. M., SCHONK, D., VERSNEL, M., STOLZ, E., and VAN JOOST, T., The detection of basal cell determinants in human psoriatic skin: a monoclonal antibody study. Arch. Dermatol. Res. 279,68-70 (1986). VERRANDO, P., and ORTONNE, J. P., Insulin binding properties of normal and transformed human epidermal cultured keratinocytes. J. Invest. Dermatol. 85, 328- 332 (1985). Authors' address: Dr. sc. med. U. WOLLlNA, Universitats-Hautklinik, Erfurter StraBe 35, Jena, DDR - 6900.
Acta histochem. 1'01.
HAHX
,"ox
Piette V
8;j
])OBSCHE, F.~LT,
an,l
ZtHLKE
VEll II tSTA v FISCHElt VEltLAG .TEXA