Tight junction components occludin, ZO-1, and claudin-1, -4 and -5 in active and healing psoriasis.

BACKGROUND: Cells of the granular layer are interconnected by tight junctions (TJs) in normal epidermis. The structural proteins of epidermal TJs include occludin, ZO-1, and claudin-1 and -4. OBJECTIVES: Our aim was to correlate the expression of TJ components with keratinocyte differentiation using psoriasis as a model of premature keratinization. METHODS: The distribution of TJ proteins was evaluated in the skin of nine patients with psoriasis. Punch biopsies were taken from perilesional skin, from active psoriasis plaques, and from healed, previously lesional locations. The punch biopsies were analysed using indirect immunolabelling for ZO-1, occludin and claudin-1, -4 and -5. In addition, epidermal samples were analysed by reverse transcription-polymerase chain reaction for claudin-1, -4 and -5 mRNAs. RESULTS: Claudin-5 was localized to the granular cell layers of normal control skin as well as perilesional and lesional psoriatic epidermis. This was unexpected, as previous studies have not detected claudin-5 in the epidermis. Occludin and ZO-1 were expressed in the granular cell layer in psoriatic perilesional epidermis. In the psoriasis plaques, ZO-1 and occludin were detected in a wider zone extending from the granular layer to the middle spinous cell layers. In healed psoriasis plaques, the expression of occludin and ZO-1 resumed a normal-looking profile, being restricted to the upper epidermis only. Claudin-1 and -4 did not show marked changes in psoriasis compared with normal skin. CONCLUSIONS: The results demonstrate claudin-5 in normal epidermis and psoriatic skin, and abnormal distribution of occludin and ZO-1 in psoriasis plaques. Clinical healing of aberrant keratinization is associated with restoration of the normal distribution of occludin, ZO-1 and also involucrin.

IgE-binding components of cultured human keratinocytes in atopic eczema/dermatitis syndrome and their crossreactivity with Malassezia furfur.

BACKGROUND: Atopic eczema/dermatitis syndrome (AEDS) patients display immunoglobulin E (IgE) reactivity to several antigens, e.g. saprophytic yeasts as Malassezia furfur. AEDS patients also show IgE autoreactivity towards cells of their own tissue including epidermis. PURPOSE OF THE STUDY: The aim of this study was to investigate the IgE autoreactivity of AEDS patients to cultured keratinocytes and to reveal potential crossreacting epitopes in cultured keratinocytes and M. furfur. MATERIAL AND METHODS: Serum samples of 27 AEDS patients were analyzed, of these 13 were M. furfur radioallergosorbent test (RAST) positive and 14 negative. Four urticaria, three psoriasis, and seven nonatopic patients were included as controls. The studies were performed by using IgE immunoblotting and immunoblotting inhibition methods. RESULTS: Ten IgE-binding protein bands were detected in cultured human keratinocytes by IgE immunoblotting using sera from adult AEDS patients. Anti-keratinocyte IgE antibodies were more associated with elevated S-IgE level than M. furfur RAST. Clear crossreactivity with M. furfur could not be shown. CONCLUSIONS: The possible pathomechanism of anti-keratinocyte IgE antibodies is not due to IgE epitope mimicry of saprophytic yeast and local tissue in AEDS skin.

Epidermal tight junctions: ZO-1 and occludin are expressed in mature, developing, and affected skin and in vitro differentiating keratinocytes.

This study demonstrates the presence of tight junction antigens in adult and developing human epidermis. Indirect immunofluorescence labeling and immunoelectron microscopy with antibodies to ZO-1 and occludin localized tight junction components ZO-1 and occludin to a narrow zone of the granular cells of adult epidermis. Double immunolabeling for tight junction components with adherens junction or desmosome proteins suggested that occludin is more specific for tight junctions than ZO-1, which may also be associated with adherens junctions. In developing skin, tight junctions interconnected the peridermal cells, and after the fetal stratification localized to the granular cell layer. Immunolabeling of psoriasis, lichen planus, and ichthyosis vulgaris, representing aberrant differentiation of the epidermis, showed that these conditions were associated with relocation of ZO-1 and occludin to the spinous cells. Cultures of epidermal keratinocytes, which offer a useful model for the formation of cellular contacts, revealed that tight junction components, ZO-1 and occludin, displayed a marked degree of colocalization relatively late during the process when the fusion zone had assumed a linear appearance. This suggests that the formation of adherens junctions and desmosomes precedes that of tight junctions. We speculate that the epidermal barrier, isolating the human body from the external environment, is in part formed by tight junctions of stratum granulosum.

Psoriasis and altered calcium metabolism: downregulated capacitative calcium influx and defective calcium-mediated cell signaling in cultured psoriatic keratinocytes.

Intracellular calcium plays an important part in the regulation of proliferation and differentiation of keratinocytes. Detached from their in vivo environment, cultured psoriatic keratinocytes were investigated by monitoring free intracellular calcium concentration, which was measured using fura-2/AM as a calcium-sensitive probe. The mean increase in intracellular calcium of psoriatic keratinocytes was significantly reduced compared with control keratinocytes when intracellular calcium stores were mobilized from endoplasmic reticulum with thapsigargin. This finding suggests defective capacitative calcium influx of psoriatic cells. Intracellular calcium stores were similar in psoriatic and control keratinocytes, when extracellular calcium was chelated with ethyleneglycol-bis(beta-aminoethyl ether)-N,N,N',N',-tetraacetic acid and intracellular calcium was depleted with thapsigargin. Mechanical wounding of keratinocyte monolayer resulted in a significantly reduced rise in intracellular calcium of psoriatic cells in low (< 0.1 mM) and high (1.8 mM) extracellular calcium suggesting defective intercellular coupling of psoriatic keratinocytes. Blocking of gap-junctions with heptanol in wounded keratinocytes did not affect the intracellular calcium response in psoriatic keratinocytes in contrast to healthy keratinocytes. Adding adenosine triphosphate to culture medium resulted in a more pronounced intracellular calcium increase than thapsigargin in psoriatic keratinocytes, suggesting that inositol triphosphate-mediated, P2-purinergic signaling was enhanced in these cells. Moreover, psoriatic keratinocytes maintained their defective responses up to at least fifth passage suggesting that psoriatic keratinocytes have an inborn error in calcium metabolism, rather than a localized defect in response to altered extracellular calcium gradient observed in vivo.