ELISA determination of sIL-15Rα, IL-15, and IL-15–sIL-15Rα proteins in sera from intact (A–C, on left) or chimeric (D–F, on right) mice stimulated with 25 μg/g poly I:C or PBS

Sera were collected 24 h after stimulation and IL-15 (B and E), sIL-15Rα (A and D), and IL-15–sIL-15Rα complex (C and F) protein levels were measured by ELISA. Each symbol reflects values obtained from individual mice.<p><b>Copyright information:</b></p><p>Taken from "IL-15Rα chaperones IL-15 to stable dendritic cell membrane complexes that activate NK cells via trans presentation"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1213-1225.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373851.</p><p></p

Effect of neutralization of IL-17 or IL-23 to ear inflammation in IMQ-treated A20 knockout mice.

<p>Thickness of IMQ- treated ears of male A20^+/- and A20^+/+ littermate mice with 100 μg/body anti-IL-17 antibody, anti-IL-12p40 antibody, or control IgG2A antibody (R&D Systems) after 6 consecutive days. Antibodies were administrated by intravenous injection the day before the IMQ treatment. Error bars represent SEM; N = 3 or 4 for each group, #<i>p</i> < 0.05 compared to IMQ- and control IgG2A-treated A20^+/+ mice and *<i>p</i> < 0.05 compared to IMQ- and anti-IL-17 antibody-treated A20^+/- mice by Student’s <i>t</i>-test.</p

Hypomorphic A20 expression confers susceptibility to psoriasis

<div><p>Psoriasis is a common inflammatory skin disease that affects approximately 1% of the population worldwide. Tumor necrosis factor-alpha-induced protein 3 (TNFAIP3) gene polymorphisms have been strongly associated with psoriasis susceptibility. In this study, we investigate how TNFAIP3, also known as A20, may regulate psoriasis susceptibility. We found that haplo-insufficient A20^+/- mice develop severe toll-like receptor (TLR)-induced skin inflammation compared to wild type mice owing to amplified production of interleukin (IL)-17 and IL-23. Examination of TNFAIP3 mRNA expression in skin biopsies from patients with psoriasis revealed reduced expression in both involved and uninvolved skin. Our results demonstrate the clinical importance of reduced dermal expression of A20 in psoriasis and suggest that A20 restriction of the IL-23/17 axis protects against psoriasis.</p></div

ELISA measurement of IL-15Rα (A) and IL-15 (B) proteins and IL-15–IL-15Rα complex (D) expression in lysates of poly I:C–stimulated BMDCs at the indicated time points

(C) ELISA determination of IL-15 protein levels after dissociation of IL-15–IL-15Rα protein complexes with 0.01% SDS and boiling. Lines represent data from WT (▪), (▴), and (▾) BMDCs. All data are representative of at least three separate experiments.<p><b>Copyright information:</b></p><p>Taken from "IL-15Rα chaperones IL-15 to stable dendritic cell membrane complexes that activate NK cells via trans presentation"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1213-1225.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373851.</p><p></p

(A) ELISA measurement of IL-12 secretion by poly I:C–stimulated BMDCs of the indicated genotypes

(B) Flow cytometric analysis of DC activation after poly I:C stimulation. Histograms of poly I:C–stimulated DCs (gray-filled histograms) of the indicated genotypes indicating surface expression levels of the surface activation markers CD40 and CD86. (C) ELISA determination of IFN-γ secretion by NK cells co-cultured with WT, 15KO, RαKO, or mixed 15KO:RαKO BMDCs. BMDCs were treated with PBS or poly I:C for 18 h, after which NK cells were co-cultured with activated DCs for an additional 6 h. Poly I:C–stimulated cultures are indicated by black bars, and PBS-stimulated control cultures are indicated by white bars. Note that significant poly I:C–induced NK cell activation occurs only in the presence of WT DCs. (D and E) Flow cytometric measurement of NK cell (NK1.1) activation by poly I:C (p(I:C)) –stimulated DC–NK cell co-cultures. Elevated CD69 surface staining reflects initial (TLR-dependent, IL-15–independent) activation of NK cells. IFN-γ and granzyme B expression by NK cells were performed by intracellular staining 6 and 12 h after stimulation, respectively. Numbers indicate the percentage of cells in the indicated gate. Note that NK cells express significant levels of both IFN-γ and granzyme B only when activated by WT DCs. Plots are representative of three separate experiments.<p><b>Copyright information:</b></p><p>Taken from "IL-15Rα chaperones IL-15 to stable dendritic cell membrane complexes that activate NK cells via trans presentation"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1213-1225.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373851.</p><p></p

A20 restricts psoriasis-related inflammatory cells responses by TLR7/8 stimulation.

<p>(A) Cytokine concentration in ears of male A20^+/- and A20^+/+ littermate mice treated with WP or IMQ for 3 or 8 consecutive days. Circles represent individual data, and horizontal bars represent the mean for each group. N = 3 or 4 for each group, *<i>p</i> < 0.05 compared to IMQ-treated A20^+/+ mice by Student’s <i>t</i>-test (B) Time course of cytokine production in splenocytes stimulated with IMQ (1 μg/mL). Splenocytes were obtained from non-treated male A20^+/- and A20^+/+ littermate mice, stimulated with IMQ for the indicated time, and the produced cytokines in the culture supernatant were mesured. Data are representative of two experiments. Error bars represent SEM; N = 3 for each group, *<i>p</i> < 0.05 compared to cytokine production in A20^+/+ splenocytes by Student’s <i>t</i>-test.</p

A20 restricts interleukin (IL)-23 signal to IL-17 producing cells.

<p>(A) Thickness of IL-23- or BSA-treated ears in A20^+/- and A20^+/+ littermate mice. Data are representative of two experiments. Error bars represent SEM; N = 3 (male, BSA injected group) or 9 (1 male and 8 female of A20^+/- mice, and 2 male and 7 female of A20^+/+ mice for IL-23-injected group) for each group,*<i>p</i> < 0.05 compared to IL-23-treated A20^+/+ mice by Student’s <i>t</i>-test. (B) Numbers of TCRγδ⁺, TCRαβ⁺, IL-17⁺ TCRγδ⁺, and IL-17⁺ TCRαβ⁺ T cells from ear-draining lymph nodes from IL-23-treated or BSA-treated mice of indicated genotypes. Data are representative of two experiments. Error bars represent SEM; N = 3 (male, BSA-injected group) or 9 (1 male and 8 female of A20^+/- mice, and 2 male and 7 female of A20^+/+ mice for IL-23-injected group) for each group, *<i>p</i> < 0.05 compared to IL-23-treated A20^+/+ mice by Student’s <i>t</i>-test.</p

Activation of IFN-γ secretion by NK cells co-cultured with various combinations of poly I:C–stimulated BMDCs and supernatants from these BMDCs

BMDCs from various genotypes (indicated along the x axis of graph) were activated with poly I:C and then supplemented with supernatants exchanged from similarly activated BMDCs. Genotypes of DCs from which supernatants were derived are indicated by individual columns (gray, WT; white, 15KO; black, RαKO; checkered, mixture of 15KO and RαKO). ELISA quantitation of NK cell IFN-γ secretion is indicated on the y axis. Note that WT DCs activate NK cells regardless of the type of supernatant added. Note also that WT DC–derived supernatants containing IL-15–sIL-15Rα complexes fail to support or augment NK cell activation.<p><b>Copyright information:</b></p><p>Taken from "IL-15Rα chaperones IL-15 to stable dendritic cell membrane complexes that activate NK cells via trans presentation"</p><p></p><p>The Journal of Experimental Medicine 2008;205(5):1213-1225.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2373851.</p><p></p

Gene expression of TNFAIP3, IL17A, IL12p40 and TNFα in psoriasis skin specimens in both non-lesional and lesional tissues.

<p>Data are obtained from GEO. DataSet type: expression profiling by array, transformed count, 82 samples, *<i>p</i> < 0.05 compared to normal skin of healthy controls and #<i>p</i> < 0.05 compared to uninvolved skin of patients with psoriasis by Steel’s test.</p

A20 expression in IECs restricts colon tumorigenesis.

<p>(A) immunoblot analysis of isolated IECs indicating efficient deletion of A20 from the small bowel (SB) and colon (C) of villin-Cre A20^FL/FL APC^min/+ mice (fl/fl) compared to control villin-Cre A20^+/+ APC^min/+ mice (+/+) mice. GAPDH is shown as a loading control. (B) Tumor number (left panel) and aggregate tumor size (right panel) in colons of A20 (fl/fl) and wild-type (+/+) mice harboring APC^min mutation. (C) Tumor numbers in small intestines of A20 (fl/fl) and wild-type (+/+) mice harboring APC^min mutation. (D) Colon and small intestine lengths from (fl/fl) and wild-type (+/+) mice harboring APC^min mutation. Each point represents one mouse. Lines indicate mean values. (f) Hematoxylin and eosin staining (upper panels) and Ki-67 and cleaved caspase-3 immunostaining (lower panels) of colonic sections from villin-Cre A20^FL/FL APC^min/+ mice (fl/fl) and control villin-Cre A20^+/+ APC^min/+ mice. 40X magnification shown.</p