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Biomedical subjects

P E van Erp

Publications and source records attributed to P E van Erp.

At least 19 recordsLinked to original sources
PubMed2001

Treatment of psoriasis with a new combination of calcipotriol and betamethasone dipropionate: a flow cytometric study.

BACKGROUND: Calcipotriol and corticosteroids are established topical antipsoriatics. Previous studies have shown that combined therapy with calcipotriol and betamethasone dipropionate was more effective than monotherapy. In the present study, a recently developed combination product of calcipotriol and betamethasone dipropionate was compared with both monotherapies and the vehicle. METHODS: Twenty-five psoriatic patients were treated twice daily with the combination product, monotherapy or vehicle during 4 weeks. Skin biopsies, taken before and after treatment, were analysed using a multi-parameter flow cytometric method. Parameters of inflammation (vimentin-positive cells), normal differentiation (keratin-10-positive cells) and proliferation (cells in SG(2)M-phase) were assessed. RESULTS: Flow cytometric analysis showed that the combination product turned out to be more effective in reducing inflammation compared with the other treatments. Restoration of normal differentiation was more advanced in patients treated with the combination product or betamethasone dipropionate compared to the vehicle. The highest number of normally differentiated cells was seen after use of the combination product. All treatments, except for the vehicle, decreased hyperproliferation. CONCLUSIONS This study shows that the combination product is a valuable new approach to the treatment of psoriasis.

Adult↗
PubMed2000

Novel functional multiparameter flow cytometric assay to characterize proliferation in skin.

Keratins are a group of cytoskeletal proteins that are found in human epidermis and other stratified squamous epithelia. Several different types of keratins have been described. Keratin 10 (K10) is a keratin that is expressed in well differentiated, suprabasal keratinocytes, and keratin 6 (K6) is a keratin which is associated with hyperproliferation. Psoriasis is a chronic inflammatory skin disease, and besides inflammation, disturbed differentiation and hyperproliferation are its hallmarks. In order to study the hyperproliferation associated keratinization in both well differentiated and poorly differentiated keratinocytes, and in order to assess the proliferative activity of all K10 and K6 subpopulations, simultaneous assessment of K6, K10, and DNA content is required. So far, a triple staining protocol had not been available. In the present study, we established a novel protocol for simultaneous measurement of K6, K10, and DNA content, which enables the characterization of the proliferative activity of several cellular subpopulations in epidermis. From 16 patients with psoriasis and from 15 healthy volunteers, punch biopsies were obtained. After preparation of single cell suspensions, cells were stained with the anti-keratin 10 IgG(1)-isotype monoclonal antibody RKSE60, with the anti-keratin 6 IgG(2a)-isotype monoclonal antibody LHK6B, and with the DNA fluorochrome TO-PRO-3 iodide. Isotype specific secondary antibodies conjugated with phycoerythrein (PE) and fluorescein isothiocyanate (FITC) were used as the second step in the staining procedure. Controls were measured omitting the primary antibodies, and gates were set in order to differentiate between the K10 and K6 subpopulations. Samples from both psoriatic patients and healthy volunteers were than measured. Owing to the IgG specificity of RKSE60 and LHK6B, no cross-reactivity was observed between these antibodies. The triple staining with RKSE60, LHK6B, and TO-PRO-3 iodide showed subpopulations of K10 expressing cells, K10/K6 co-expressing cells, and K6 only expressing cells. There was a significant difference in the proportion of K6 expression and K10/K6 co-expression between psoriatic and normal skin. Moreover, the proliferative activity of these subpopulations could be quantified by this protocol. We concluded that a triple staining protocol for the assessment of K6, K10, and DNA content, using the monoclonal antibodies LHK6B, RKSE60, and TO-PRO-3 iodide, supplies reliable and reproducible data for cellular studies on these keratins and for studying the proliferative activity of the subpopulations of these keratins in epidermis. Moreover, the present study showed that with respect to the proportion of K6, significant differences are present between psoriatic and healthy human skin.

Adult↗
PubMed2000

Coexpression of keratins 13 and 16 in human keratinocytes indicates association between hyperproliferation-associated and retinoid-induced differentiation.

In human skin, epidermal differentiation occurs in two ways: normal differentiation, characterized by keratin 10 expression, and alternative differentiation. Alternative differentiation can be hyperproliferation-associated differentiation (expression of keratin 16) or the reinduction of an embryonic type of differentiation (expression of keratin 13). This embryonic type of differentiation is also seen following treatment with retinoids. In the present study, the hypothesis that hyperproliferation-associated and retinoid-induced differentiation are separate processes was investigated. Two areas of normal skin were treated for 24 h with 0. 1% all-trans-retinoic acid. Subsequently, one of the areas was tape-stripped and treatment was continued for 48 h. Multiparameter flow cytometry permitted simultaneous measurement of two coexpressed differentiation markers (retinoid-induced and normal differentiation or retinoid-induced and hyperproliferation-associated differentiation) and the proliferation characteristics (cells in S/G(2)M phase). Concerning normal and retinoid-induced differentiation, the all-trans-retinoic acid-induced expression of keratin 13 was only seen in tape-stripped retinoid-treated skin and exclusively together with that of keratin 10. The assessment of hyperproliferation-associated and retinoid-induced differentiation showed slight expression of keratin 13 without expression of keratin 16 in tape-stripped skin. Coexpression of keratins 16 and 13 was exclusively seen in tape-stripped retinoid-treated skin. The finding that keratin 13 expression following treatment with all-trans-retinoic acid occurred exclusively in hyperproliferative skin suggests that retinoid-induced and hyperproliferation-associated differentiation are coupled. Coexpression of keratins 13 and 16 provides direct experimental evidence for this association.

Adhesives↗
PubMed2000

Flow-cytometric characterization of normal versus psoriatic epidermis using improved cell separation methodology.

Recently a new approach for epidermal cell characterization was developed: three-parameter flow cytometrical analysis of pure and complete epidermal cell suspensions prepared from punch biopsies followed by dermoepidermal separation by thermolysin. The aim of the present communication is the comparison between psoriatic lesional skin and normal skin using this new approach with respect to the percentage of suprabasal keratinocytes (keratin 10+ cells), mesenchymal cells, including the infiltrate cells (vimentin+ cells) and the percentage of basal cells in SG2 M phase, in order to validate this methodology in studies on psoriatic skin. Punch biopsies were taken from 7 healthy volunteers and in 7 psoriatic patients 4 biopsies were taken in each of them from comparable lesions. The present study reconfirmed that the percentage of basal keratinocytes in psoriasis was increased and the percentage of keratin 10+ cells was substantially decreased as compared to normal skin. The new methodology revealed data with a narrow range. In psoriatic lesional skin the intra individual variation was less compared to the inter individual variation.

Case-Control Studies↗
PubMed2000

Efficacy of calcipotriol ointment applied under hydrocolloid occlusion in psoriasis.

BACKGROUND: Hydrocolloid (HCD) dressings enhance the efficacy of topical corticosteroids. OBJECTIVE: We wanted to evaluate the effect of calcipotriol ointment under an HCD dressing in the treatment of psoriatic plaques. METHODS: In 9 psoriatic patients, we cleared one plaque using this approach and took biopsies at start, clearance and relapse. Clinical and immunohistochemical validation was assessed. RESULTS: After an average treatment of 3.6 weeks, each lesion had cleared (apart from some residual erythema). The average remission period was 8 weeks. During this treatment, the number of cycling epidermal cells (Ki-67-positive nuclei) and the expression of keratin 14 and keratin 16 had decreased substantially. In biopsies taken from the skin immediately adjacent to the relapsing lesion, these markers remained reduced which indicated the prolonged effect of calcipotriol on epidermal differentiation. CONCLUSION: It is speculated that combination therapy of calcipotriol with treatments with a different mode of action such as photo(chemo)therapy, corticosteroids and cyclosporine might be worthwhile.

Adult↗
PubMed2000

Changes in keratin 6 and keratin 10 (co-)expression in lesional and symptomless skin of spreading psoriasis.

BACKGROUND: Keratin 6 (K6) and keratin 10 (K10) are markers for epidermal hyperproliferation and differentiation, respectively, and are both expressed in the suprabasal layers of the epidermis. They may be co-expressed in different stages of the spreading psoriatic lesion, but single expression can also occur. OBJECTIVE: To investigate to what extent keratinocytes express K6 and K10, and to what extent they co-express K6 and K10 in different stages of the psoriatic lesion. We studied this in spreading psoriatic plaques. METHODS: Three 3-mm punch biopsies were obtained from the inner involved margin of a spreading lesion, from the uninvolved skin immediately adjacent to the spreading plaque, and from the distant uninvolved skin of 8 patients with incipient psoriasis. From 9 healthy volunteers, 3-mm punch biopsies were obtained as controls. After preparation of single cell suspensions of these biopsies, a triple staining protocol was performed with markers for K6 (monoclonal antibody LHK6B), K10 (monoclonal antibody RKSE60) and DNA content (TO-PRO-3 iodide). Subsequently, cells were measured with a flow cytometer and the proportion of the markers was calculated using specific software. RESULTS: We observed a population of K6/K10-co-expressing cells, but also populations expressing only K6. These subpopulations varied with the involvement of the lesion. There was a statistically significant difference between the inner margin and the outer margin with respect to the proportion of K6- and K10-expressing cells, whereas more K6-positive and K10-negative cells were detected in the inner margin of the lesions. The proportion of K6/K10-co-expressing cells in the inner margin was significantly different from the distant uninvolved skin. CONCLUSION: We confirmed that individual keratinocytes in psoriasis can express K6 or K10 depending on their localization in involved or uninvolved skin. There is a unique subpopulation of cells in the psoriatic plaques which co-express K6 and K10. More studies are required to fully understand the pathogenic relevance of co-expression and single expression of K6 and K10.

Adult↗
PubMed2000

Cell-kinetic evidence for increased recruitment of cycling epidermal cells in psoriasis: the ratio of histone and Ki-67 antigen expression is constant.

BACKGROUND: One of the hallmarks of the psoriatic plaque is increased epidermal proliferation. Whether this is the result of an increased recruitment of cycling epidermal cells or a decrease in cell cycle time has been a matter of debate for years. OBJECTIVE: Calculating cell-kinetic information from the number of S phase cells in psoriasis by in situ hybridisation using a histone probe and the number of cycling epidermal cells by immunohistochemistry using the MIB-1 antibody. METHODS: Immunohistochemistry and non-isotopic in situ hybridisation were performed on serial sections of 33 untreated psoriatic samples and 14 tacalcitol-treated samples. RESULTS: The labelling index (number of cells in S phase/number cycling cells per millimetre length of section) in psoriatic untreated as well as in treated plaques is 16%. The amount of S phase cells in our experiment is equal compared with the number of cells in S phase as determined by BrdU incorporation. CONCLUSION: Using this direct approach to study cell-kinetic behaviour of psoriatic skin, we reconfirm that the psoriatic abnormality is due to a defect in the G(0)-G(1) recruitment mechanism (by increased recruitment of G(0) cells), a decrease in apoptosis or an increase in the number of cell divisions in the transit-amplifying compartment, rather than a reduction in the cell cycle time.

Cell Cycle↗
PubMed1999

The epidermal phenotype during initiation of the psoriatic lesion in the symptomless margin of relapsing psoriasis.

BACKGROUND: The mature psoriatic lesion does not necessarily demonstrate changes relevant to early phases of the lesion. OBJECTIVE: In a model for relapsing psoriasis we examined the epidermal phenotype by means of a panel of immunohistochemical parameters: keratins 14 and 16, epidermal growth factor receptor (EGFR), Ki-67 antigen, and Tdt-mediated Unscheduled Nick End Labeling to detect apoptosis. METHODS: In 9 patients, we cleared psoriatic plaques by topical treatment with clobetasol-17-propionate under hydrocolloid occlusion. Relapse (defined as a clinical sum score > or = 6) was awaited. Biopsy specimens of the psoriatic lesion, the cleared skin, the relapsed plaque, and its clinically normal margin were assessed. RESULTS: Psoriasis recurred after 19+/-6 weeks (mean +/- SEM). During treatment all parameters improved considerably; however, the number of apoptotic cells was not affected. Ki-67 values decreased well below the normal range. At initial relapse, the symptomless skin adjacent to the relapsing lesion (margin) showed a marked expression of keratin 16 and EGFR. Ki-67 expression was increasing in the margin but was below values of the mature lesion. The localization of cycling cells in the first suprabasal layers was a remarkable feature. Keratin 14 expression was increased in the recurrent lesion itself, but not in the symptomless margin. CONCLUSION: Keratin 16 and EGFR expression are early phenomena in the evolution of the lesion, and they anticipate epidermal proliferation. The expression of keratin 14 follows overt epidermal hyperproliferation. The present observation in incipient psoriasis lends support to the hypothesis that the basal cell compartment does not have a primary involvement in the initiation of epidermal abnormalities in psoriasis, but that a coordinated sequence of events involving proliferation and differentiation markers in the first suprabasal layers of the epidermis could be the key to the pathogenesis of this puzzling disease.

Administration, Topical↗
PubMed1999

Clobetasol under hydrocolloid occlusion in psoriasis results in a complete block of proliferation and in a rebound of lesions following discontinuation.

BACKGROUND: It is a well-established fact that very potent corticosteroids are highly effective in the treatment of psoriatic plaques, and that the addition of a hydrocolloid occlusive dressing (HCD) enhances its efficacy. It is known that topical steroids induce normal differentiation and diminish hyperproliferation during treatment of psoriatic plaques. These changes are reflected by an increase in keratin 10 (K10) and a decrease in keratin 6 (K6), respectively. However, the influence of class IV corticosteroids under HCD on K10 and K6 subpopulations has never been studied. OBJECTIVE: The present study was performed to study quantitatively the influence of a class IV topical steroid under HCD on the dynamics of K10 and K6 subpopulations. METHODS: In the present study, we treated moderately severe stable psoriatic plaques with clobetasol-17-propionate under HCD until clearance was achieved. We took biopsies prior to treatment, after clearance and at relapse. Using flow cytometry, we studied the dynamics of K10 and K6 subpopulations. RESULTS: Prior to treatment, 41.8% of all cells expressed K6. After treatment-induced clearance, this proportion decreased to 1.2%, which is below the normal range. At relapse, pre-treatment levels were observed again. A trend to an increasing number of basal cells and an increase in the proliferative activity of these basal cells at relapse compared to the stable situation prior to treatment was observed. CONCLUSION: We conclude that clobetasol under hydrocolloid occlusion induces virtually a total block of proliferation of the basal cell population and decreases hyperproliferation-associated keratins dramatically. Furthermore, based upon epidermal cell characteristics, we conclude that a rebound phenomenon occurs following discontinuation of therapy with clobetasol under hydrocolloid occlusion.

Administration, Topical↗
PubMed1999

Sequence-specific inhibition of gene expression in intact human skin by epicutaneous application of chimeric antisense oligodeoxynucleotides.

Targeted and selective inhibition of keratinocyte gene expression in human epidermis could be an efficient and safe pharmacologic approach in many skin diseases. In this study we investigated whether topical application of antisense oligodeoxynucleotides (ODN) on intact human skin can be used to inhibit expression of a gene in the differentiated compartment of the epidermis. We applied a variety of 20-mer antisense and control ODN designed to hybridize to different regions on the mRNA of the inducible epidermal proteinase inhibitor skin-derived antileukoproteinase (SKALP)/elafin that was used as a model target gene. When nuclease-resistant fully phosphorothioate ODN were applied to explant cultures of human skin, they were found to be either ineffective at low doses or severely toxic at higher doses which could be attributed to the extremely high degree of protein binding found with this type of ODN. When chimeric ODN with a phosphodiester core and phosphorothioate 5' and 3' ends were applied to intact skin, no toxicity was noted. One of the tested chimeric ODN, that exhibit only minor protein binding, was found to inhibit SKALP expression at the protein level in a dose-dependent manner. The observed inhibition on SKALP expression levels was specific as evaluated by application of strict criteria. Sequence specificity was assessed by the addition of sense and scrambled ODN which were ineffective. Furthermore the expression levels of three other differentiation-related genes (involucrin, cytokeratin 16, and secretory leukocyte proteinase inhibitor) were not affected, indicating that the inhibition was gene specific. Confocal laser scanning analysis of fluorescently labeled ODN confirmed that these molecules can easily penetrate the epidermis and localize in the cytoplasm of differentiated keratinocytes. We conclude that topical application of antisense ODN can be used to modulate epidermal gene expression, and could potentially be useful to inhibit expression of genes that are relevant in skin diseases.

Absorption↗
PubMed1998

Flow cytometric and microscopic characterization of the uptake and distribution of phosphorothioate oligonucleotides in human keratinocytes.

Gene-specific inhibition by antisense oligonucleotides has been successful in a large number of systems. In an attempt to use this strategy for the modulation of skin disease-specific gene expression, we studied oligonucleotide uptake in cultured human keratinocytes. This study revealed a heterogeneous uptake of fluorescently labeled phosphorothioate oligonucleotides. Flow cytometric and microscopic analysis showed two fluorescent cell populations with differences in intensity: a 'bright' population of highly fluorescent small cells and a 'dim' population of less fluorescent but larger cells. The heterogeneity in uptake between these two populations was not a result of differences in cell cycle phases of the keratinocytes, as shown by flow cytometric sorting and measurements of relative DNA content. In both populations the oligonucleotides were transported intracellularly and were mainly located in the cytoplasm. A typically speckled localization pattern was demonstrated by confocal laser scanning microscopy. We used propidium iodide (PI) to assess viability, and showed that in nonviable (PI-permeable) keratinocytes the oligonucleotides accumulated in the nucleus. The use of a lipidfection reagent also changed the intracellular distribution of oligonucleotides from a punctate cytoplasmic pattern to an intense nuclear localization. The process of uptake by the viable keratinocytes was dependent on oligonucleotide concentration, incubation time and temperature. This study underlines the importance of kinetic studies on oligonucleotide uptake in human keratinocytes which must be considered when specific oligonucleotides are used against skin disease-specific genes.

Base Sequence↗
PubMed1998

The epidermis of chronic idiopathic lichen planus during topical treatment with the vitamin D3 analogue KH 1060.

A parallel-group, double-blind, randomised study was performed to establish the effect of the vitamin D3 analogue KH 1060, in an ointment versus vehicle only, on the epidermal cell characteristics of chronic idiopathic lichen planus; KH 1060 also has marked immunosuppressive activity. A group of 10 patients were treated for 8 weeks with either KH 1060 ointment or vehicle only. In addition to the assessment of clinical scores, keratotome biopsies were taken before and after 8 weeks' treatment. Epidermal cell suspensions were prepared with trypsin and the suspensions incubated with TO-PRO-3 (DNA marker), RKSE 60 (marker for keratin 10-positive cells) and antivimentin (marker for all non-keratinocytes). In nine of the 10 patients, keratotome biopsies were obtained both before and after 8 weeks treatment. The vehicle alone had no significant effect on the clinical severity scores or epidermal cell characteristics. In contrast, the KH 1060 ointment resulted in a statistically significant reduction in the percentage of cells in S- and G2M phase and the percentage of vimentin-positive cells, but it did not affect the percentage of keratin 10-positive cells.

Calcitriol↗
PubMed1998

A clinical and flow cytometric model to study remission and relapse in psoriasis.

The aim of the present study has been to analyse remission and relapse characteristics in psoriasis vulgaris. In 15 patients, two different psoriatic lesions (clinical and flow cytometric study) were treated with clobetasol propionate until clearance for maximally 23 days. In the clinical study only cleared lesions were divided into three test sectors with different post-clearance treatment: (1) alcoholic solution under occlusion, (2) occlusion only, and (3) no treatment. In the flow cytometric study, biopsies were taken from the test lesion before clobetasol therapy (i), at clearance (ii), and at relapse from both visibly affected and unaffected skin (iii, iv). Epidermal proliferation, differentiation and inflammation were quantified by multiparameter flow cytometry. The clinical evaluation worked well and could discriminate between the different therapy modalities. After 28 days, 80% of untreated sectors showed a relapse. Occlusion decreased this percentage to 50%. Application of the alcoholic solution further decreased this percentage to 30%. The flow cytometric analysis demonstrated a very low proliferative activity of the basal compartment at clearance. This activity was higher in the visibly unaffected skin at relapse, whereas highest values were assessed in the affected skin at relapse. Interestingly, at relapse the proliferative activity in the suprabasal compartment of the visibly unaffected skin had increased to values identical to the affected skin. The present model allows standardized comparison of different approaches for maintenance therapy in psoriasis vulgaris. We demonstrate that occlusion has an inhibitory effect on the tendency to relapse after successful treatment with clobetasol propionate. Quantitative information on remission and relapse of psoriasis can be obtained by multiparameter flow cytometry.

Administration, Topical↗
PubMed1998

Multi parameter flow cytometric assessment of regenerative epidermis with special reference to the antiproliferative effect of occlusion.

Multi parameter flow cytometrical assays permit simultaneous assessment of proliferation, differentiation, and inflammation parameters. In this study, the validation of TO-PRO-3 iodide (TP3) compared to propidium iodide (PI) and DE-K10 compared to RKSE60 were evaluated in tape stripping induced hyperproliferation. No occlusion, Duoderm (intermediate occlusion) and Blenderm (maximal occlusion) were used as a model to evaluate the effect of occlusion on epidermal regeneration. Proliferation in the keratin 10-negative compartment measured with TP3 proved to be a good approximation of proliferation measured with PI. Other epidermal subpopulations (keratin 10-dim and -bright cells) did not make a relevant contribution to hyperproliferation. DE-K10 is probable more sensitive than RKSE60 to distinguish populations that differ in degree of differentiation. Occlusion of tape stripped skin resulted in decreased proliferation and increased differentiation. This effect was most pronounced with maximal occlusion. This study showed that occlusion is a therapy, which realises normalisation of hyperproliferative skin disorders.

Biopsy↗
PubMed1997

The regulation of CD11b integrin levels on human blood leukocytes and leukotriene B4-stimulated skin by a specific leukotriene B4 receptor antagonist (LY293111).

CD11b is part of the beta2-integrin Mac-1 and plays an important role in neutrophil adhesion. Leukotriene B4 (LTB4) is an active upregulator of neutrophil CD11b-expression, acts as a potent chemoattractant to neutrophils and is also known to upmodulate epidermal proliferation. We performed a placebo-controlled study on LY293111, an oral LTB4 receptor antagonist. Twenty healthy male volunteers were randomised over three treatment groups that received placebo, 48 mg, or 200 mg drug twice daily for 10 days. Before and after treatment, flow cytometrical CD11b assessment was performed on in vitro LTB4-stimulated peripheral blood neutrophils. Additionally, skin biopsies were taken at 24 and 72 h after epicutaneous LTB4 application, before and after treatment. The effects on skin were assessed immunohistochemically using various markers. All observed effects were dose related. CD11b upregulation on blood neutrophils was significantly suppressed in both treatment groups compared to placebo. In skin, a significant suppression of inflammation and hyperproliferation occurred. Pronounced inhibition was observed on neutrophil migration into the epidermis and the inflammatory infiltrate was decreased. A similar but weaker response was seen in the dermis. The number of cycling cells as well as suprabasal keratin-16 expression were decreased in both treatment groups. LY293111 proved to be a potent inhibitor of LTB4-induced cutaneous inflammation and hyperproliferation. The potent antiinflammatory effect in vivo and the fact that in the present study the compound showed no clinically significant side effects make it an interesting drug in the future treatment of inflammatory conditions predominated by neutrophils.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed1997

Multiparameter flow cytometry as a tool to evaluate antipsoriatic therapy.

Objective comparison of different antipsoriatic therapies requires quantitative assessment of disease severity. However, clinical assessment with the widely used Psoriasis Area and Severity Index (PASI) introduces inaccuracy. An alternative is the quantitative analysis of different epidermal cell parameters using multiparameter flow cytometry. Our aim in the present study was to compare the clinical and flow cytometric approach to monitor disease activity and to evaluate antipsoriatic efficacy. Clinical scores for erythema, induration and scaling were assessed and biopsies for flow cytometric analysis were obtained from the psoriatic plaques of 89 patients before and after treatment with different therapeutic regimens consisting of vitamin D3 analogues and corticosteroids. In total, 219 epidermal cell suspensions were analysed using triple-labelling, with the simultaneous staining of markers for epidermal proliferation (DNA dye TO-PRO-3), differentiation (antikeratin 10), and inflammation (antivimentin). Correlation analysis was performed on 166 paired values obtained from 83 patients. A highly significant correlation was observed between erythema and the percentage of vimentin-positive cells, between scaling and the percentage of keratin 10 positive keratinocytes, and between induration and the number of basal keratinocytes in the S- and G2M phase, when all 166 biopsies were assessed. The correlation remained in the same range if the analysis was restricted to the 83 pretreatment biopsies. In contrast to the clinical scores, the flow cytometric analysis permitted a clear separation between the antiproliferative and anti-inflammatory or keratinization-enhancing effects of antipsoriatic treatment. The vitamin D3 analogues proved to exert a mainly antiproliferative effect. The combination of calcipotriol and a topical corticosteroid improved all cell biological markers substantially, and clobetasol monotherapy had a powerful effect on these markers. In conclusion, multiparameter flow cytometry has been shown to be a sensitive tool to evaluate the growth inhibiting, anti-inflammatory and keratinization-enhancing effects of antipsoriatic therapies.

Biomarkers↗