Multiparameter flow cytometry as a tool to evaluate antipsoriatic therapy

Glade, C.P.; Erp, P.E.J. van; Boezeman, J.B.M.; Kerkhof, Peter C M van de

doi:10.1111/j.1365-2133.1997.tb03740.x

Cited by 13 publications

(15 citation statements)

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“…In order to quantify the severity of psoriatic plaques at the cellular level, a flow cytometric triple parameter method has been developed by Van Hooijdonk et al (43), and Glade et al (17) using markers for keratin 10 (RKSE60), mesenchymal cells (vimentin), and DNA content (TO‐PRO‐3 iodide). These three parameters are assessed in epidermal cell suspensions and have proven to correlate highly significant with the clinical parameters for scaling, erythema, and plaque thickening, respectively, as represented by the Psoriasis Area and Severity Index (PASI (14)) (17). They have proven to be very useful in quantifying the efficacy of antipsoriatic drugs (18, 19, 30).…”

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confidence: 99%

“…The aim of the present study was to establish a new flow cytometric protocol with simultaneous staining for keratin 1/10, keratin 6/16, and DNA content, in order to be able to assess the dynamics of all keratin 1/10 and keratin 6/16 subpopulations, and to assess the proliferative activity of these subpopulations. In particular, we tried to adapt the protocol of Van Hooijdonk et al (43) and Glade et al (17) by replacing the inflammation marker vimentin by a hyperproliferation marker.…”

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Novel functional multiparameter flow cytometric assay to characterize proliferation in skin

2000

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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 IgG1‐isotype monoclonal antibody RKSE60, with the anti‐keratin 6 IgG2a‐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.Cytometry (Comm. Clin. Cytometry) 1:43–49, 2000 © 2000 Wiley‐Liss, Inc.

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Novel functional multiparameter flow cytometric assay to characterize proliferation in skin

2000

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“…Epidermal thinning was observed as a result of clobetasol while calcipotriol led to increased thickness of the epidermis. Glade et al 10 . used flow cytometric methods to differentiate between the anti‐inflammatory, growth‐inhibiting and keratinization‐enhancing properties of several antipsoriatic therapies.…”

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Bioavailability of betamethasone dipropionate when combined with calcipotriol

Traulsen

2004

Int J Dermatology

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“…Calcipotriol is a synthetic vitamin D 3 analogue for the topical treatment of chronic plaque-stage psoriasis. Calcipotriol induces differentiation and inhibits proliferation of keratinocytes both in vitro and in vivo coupled with immunomodulatory activity [3, 4, 5]. Several large-scale clinical studies have demonstrated the safety and efficacy of treatment with calcipotriol either in ointment or cream formulation [6, 7, 8, 9, 10, 11].…”

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Calcipotriol Solution for the Treatment of Scalp Psoriasis: Evaluation of Efficacy, Safety and Acceptance in 3,396 Patients

et al. 2001

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Background: Psoriasis of the scalp is a very common disease, cosmetically disturbing and therapeutically difficult to manage. Objective: The aim of our study was to investigate the efficacy, safety and cosmetic acceptance of calcipotriol solution in a large number of patients with mild to moderate scalp psoriasis and to compare this treatment with previous therapies used. Methods: In this multicentre prospective observational cohort study 3,396 patients were treated with calcipotriol solution (50 µg/ml) twice daily over an 8-week period either alone or in combination with other treatments. The psoriasis scalp severity index (PSSI) and investigator/patient global assessment were used for the evaluation of clinical response. Results: All psoriasis severity parameters measured were reduced with a significant decrease in PSSI scores from 18.4 to 5.6 after 8 weeks of therapy (p < 0.001). About 80% of the patients showed very good or good clinical improvement. Combination of calcipotriol solution with other treatment modalities e.g. corticosteroids or salicylic acid, showed an increased treatment response. In only 2.4% of the patients side effects occurred (e.g. irritation). Conclusion: Calcipotriol solution is an effective, safe, well-tolerated and cosmetically acceptable treatment modality. By patients and physicians, this treatment was found to be a valuable supplement to previously available and established treatments for scalp psoriasis.

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Multiparameter flow cytometry as a tool to evaluate antipsoriatic therapy

Cited by 13 publications

References 13 publications

Novel functional multiparameter flow cytometric assay to characterize proliferation in skin

Novel functional multiparameter flow cytometric assay to characterize proliferation in skin

Bioavailability of betamethasone dipropionate when combined with calcipotriol

Calcipotriol Solution for the Treatment of Scalp Psoriasis: Evaluation of Efficacy, Safety and Acceptance in 3,396 Patients

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