Article Figures & Data
Figures
- FIGURE 1.
Immunohistochemical staining of CD4, CD8, neutrophil elastase, and CXCL8 in normal skin and inflamed tissue. In normal skin (A and B) and in atopic dermatitis (E and F), a low number of CD4+ and CD8+ cells was observed compared with pustular psoriasis (I and J, patient PP3), Behçet’s disease (M and N, patient BD1), and AGEP (O and R, patient AG2). The majority of the T cells were located in the dermis, and most of them were CD4+. Neutrophil elastase was absent in normal skin (C) and barely detectable in atopic dermatitis (D), whereas a high number of PMN could be detected in all autoinflammatory diseases (K, O, and S) situated characteristically within the subcorneal pustules where the PMN are the dominant cell type. CXCL8 was barely detectable in normal skin (D) and in atopic dermatitis (H). In pustular psoriasis (L) and in AGEP (T), CXCL8+ cells were distributed in the entire epidermis and in dermal infiltrates. In Behçet’s disease (P), CXCL8+ cells were augmented and mainly located in the basal and suprabasal layers of the epidermis and to a lesser degree in the dermis. Original magnifications, ×100.
- FIGURE 2.
Quantification of CD4+, CD8+, and CXCL8+ cells in normal skin, atopic dermatitis, and inflamed skin. Comparison of CD4+ (A), CD8+ (B), and CXCL8+ cells (C) in dermis of normal skin (NS; n = 2), atopic dermatitis (AD; n = 2), patient PP3 (pustular psoriasis), patient BD1 (Behçet’s disease), BD3 (Behçet’s disease), and patient AG2 (AGEP). The highly significant differences of CXCL8+ cells in normal skin/atopic dermatitis vs pustular psoriasis, Behçet’s disease, and AGEP are indicated. Mean and SEM of different sections is given (n = 8–12 fields per patient; Kruskal-Wallis nonparametric ANOVA test: ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001).
- FIGURE 3.
Double staining of CD4 and CXCL8 expression in psoriatic skin (patient PP3). Single-positive CD4+ cells (red staining, red arrow) and CXCL8+ cells (brown staining, yellow arrow) are shown. The presence of CD4+CXCL8+ T cells was demonstrated with red and brown double-stained cells (black arrow). Original magnification, ×400.
- FIGURE 4.
Expression of CCR6 and its ligand CCL20 in normal skin and in inflamed tissue. CCR6 is only scarcely expressed in normal skin (A) and in atopic dermatitis (C). In pustular psoriasis (E, patient PP3), Behçet’s disease (G, patient BD1), and AGEP (I, patient AG2), an increased expression of CCR6+ cells can be seen, predominantly in the basal layer epidermis and around the vessels in the dermis. Compared with normal skin (B) and atopic dermatitis (D), CCL20 expression was enhanced in pustular psoriasis (F), Behçet’s disease (H), and in AGEP (J). Original magnifications, ×100.
- FIGURE 5.
Flow cytometric detection of CCR8 and CCR6 expression in TCC eluted from inflamed skin. TCC eluted from pustular psoriasis (rectangles; n = 11; TCC from two patients), Behçet’s disease (triangles; n = 12; one patient), and AGEP (circles; n = 13; two patients) were analyzed for CCR8 (A) and CCR6 (B) expression. Most of the clones expressed CCR6, whereas CCR8 was absent in the clones eluted from inflamed tissue (22 ). Mean values are indicated as horizontal bars.
- FIGURE 6.
Characterization and cytokine/chemokine secretion of skin-derived CD4+ TCRab+ TCC from patients with pustular psoriasis, Behçet’s disease, AGEP, and from normal skin. TCC of patients PP1, PP2, and PP4 with pustular psoriasis (A), of patient BD1 with Behçet’s disease (B), patients AG2 and AG3 with AGEP (C), and from two different donors of normal skin NS1, NS2 (D) were stimulated with anti-CD3/anti-CD28 Abs, and the cytokine secretion patterns were determined. The differences between stimulated and unstimulated values are given. The mean cytokine/chemokine secretion of the unstimulated cells was always <0.2 ng/ml, except for CXCL8 in TCC from patient BD1 (unstimulated levels of CXCL8 0 −2.1 ng/ml, mean: 0.5 ng/ml). Cytokine (IL-4, IL-5, IFN-γ, TNF-α, GM-CSF) and chemokine (CXCL8) levels are shown as dot plots, where each symbol represents one TCC. TCC are grouped into different cytokine secretion patterns: Th1-like (gray open symbols), Th2-like (gray filled symbols), Th0-like (black open symbols), predominantly CXCL8/GM-CSF-producing T cells (black filled symbols), and “low producers” (black-gray symbols).
- FIGURE 7.
Composition of T cells contributing to CXCL8/GM-CSF production and neutrophilic inflammation in the skin. In healthy individuals, CCR8+ skin homing T cells have the ability to produce CXCL8, after stimulation only (illustrated by dashed line). In acute neutrophilic inflammations, CXCL8 production occurs mainly by skin-infiltrating CCR6+ T cells, which also produce moderate amounts of TNF-α, IFN-γ, and IL-4/IL-5 (Th0) or are already biased to produce a more restricted set of cytokines (Th1, occasionally Th2). In chronic relapsing neutrophilic inflammations, CXCL8/GM-CSF derive from various CCR6+ T cells, some of which produce mainly CXCL8/GM-CSF, but not other cytokines (ThCXCL8). The strength of the arrows reflects the relative contribution of the various subsets.
Tables
- Table I.
Characteristics of patients with Behçet’s disease (BD1-BD3), pustular psoriasis (PP1-PP4), and AGEP (AG1-AG2)a
Patients Disease Age (years)/Sex (M/F)/Disease since Drug Intake PP1 Pustular psoriasis 59/F/unknown Tacrolimus, flumetasone, + salicylic acid (topical) naproxen, metoprolol, + irbesartan (oral) PP2 Pustular psoriasis 69/F/53 years Betamethasone (topical), UV PP3 Pustular psoriasis 46/F/4 years (recurrent) Prednisolone (discontinued 5 days before biopsy) PP4 Pustular psoriasis 36/M/3 wk Infliximimab (last injection 1 mo ago) BD1 Behçet’s disease 44/M/4 years Prednisone BD2 Behçet’s disease 31/M/5 years Acetylsalicylic acid BD3 Behçet’s disease 44/M/8 years Leflunomide, colchicine AG1 AGEP 38/F/acute Hypersensitivity to nickel (screw for orthopedic surgery) AG2 AGEP 43/M/acute Hypersensitivity to prednisone and hydrocortisone AG3b AGEP 30/F/cured Hypersensitivity to amoxicillin -
a PP, pustular psoriasis; BD, Behçet’s disease, AG, AGEP.
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b Previously characterized as patient AP (8 ), cells obtained from positive patch test reaction.
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Disease No. of TCC Tested (n) Th1-Like Cytokine Secretion Pattern Th2-Like Cytokine Secretion Pattern Th0-Like Cytokine Secretion Pattern Predominant CXCL8 and GM-CSF Secretion Low Producers IFN-γ > 8 ng/ml IL-5 > 1 ng/ml IFN-γ > 1 ng/ml and IL-5 > 0.2 ng/ml together Non-Th1, Th2, Th0 but CXCL8 > 0.2 ng/ml and GM-CSF > 0.2 ng/ml Non-Th1, Th2, Th0, and no CXCL8 production Total CXCL8 (−/+/++) Total CXCL8 (−/+/++) Total CXCL8 (−/+/++) Total CXCL8 (−/+/++) Total CXCL8 (−/+/++) Pustular psoriasis 13 2 (1/0/1) 2 (1/0/1) 2 (2/0/0) 4 (0/3/1) 3 (3/0/0) Behçet’s disease 13 5 (1/0/4) 1 (0/1/0) 5 (0/2/3) 2 (0/0/2) 0 AGEP 10 4 (2/0/2) 1 (1/0/0) 4 (0/4/0) 0 1 (1/0/0) Normal skin 15 6 (0/3/3) 0 7 (2/4/1) 0 2 (2/0/0) -
a TCC were grouped into Th1-like (IFN-γ > 8 ng/ml), Th2-like (IL-5 > 1 ng/ml), and Th0-like cells (IFN-γ > 1 ng/ml together with IL-5 > 0.2 ng/ml). Predominantly CXCL8/GM-CSF-producing T cells do not belong to Th1 (<8 ng/ml IFN-γ), nor to Th2 (<1 ng/ml IL-5), nor to Th0 (no simultaneous IFN-γ and IL-5 production). They are characterized by medium to high CXCL8 (>0.2 ng/ml) production together with GM-CSF (>0.2 ng/ml). “Low producers” secreted low amounts of all cytokines/chemokines. CXCL8-producing TCC are shown in each group (−, <0.2 ng/ml; +, 0.2–1 ng/ml; ++, >1 ng/ml). Details are shown in Fig. 6⇑.
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