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CARD14 expression in dermal endothelial cells in psoriasis.

Harden JL, Lewis SM, Pierson KC, Suárez-Fariñas M, Lentini T, Ortenzio FS, Zaba LC, Goldbach-Mansky R, Bowcock AM, Lowes MA - PLoS ONE (2014)

Bottom Line: CARD14 is a scaffolding protein that regulates NF-κB activation, and psoriasis-associated CARD14 mutations lead to enhanced NF-κB signaling.CARD14 is expressed mainly in epidermal keratinocytes, but also in unidentified dermal cells.Transfection of dermal ECs with psoriasis-associated CARD14 mutations resulted in increased expression of several chemokines, including CXCL10, IL-8, and CCL2.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Investigative Dermatology, The Rockefeller University, New York, New York, United States of America.

ABSTRACT
Mutations in the caspase recruitment domain, family member 14 (CARD14) gene have recently been described in psoriasis patients, and explain the psoriasis susceptibility locus 2 (PSORS2). CARD14 is a scaffolding protein that regulates NF-κB activation, and psoriasis-associated CARD14 mutations lead to enhanced NF-κB signaling. CARD14 is expressed mainly in epidermal keratinocytes, but also in unidentified dermal cells. In this manuscript, the identity of the dermal cell types expressing CARD14, as well the potential functional consequence of overactive CARD14 in these dermal cell types, was determined. Using two-color immunofluorescence, dermal CARD14 did not co-localize with T-cells, dendritic cells, or macrophages. However, dermal CARD14 did highly co-localize with CD31(+) endothelial cells (ECs). CARD14 was also expressed non-dermal endothelial cells, such as aortic endothelial cells, which may indicate a role of CARD14(+)ECs in the systemic inflammation and cardiovascular comorbidities associated with psoriasis. Additionally, phosphorylated NF-κB was found in psoriatic CARD14(+) CD31(+) ECs, demonstrating this pathway is active in dermal ECs in psoriasis. Transfection of dermal ECs with psoriasis-associated CARD14 mutations resulted in increased expression of several chemokines, including CXCL10, IL-8, and CCL2. These results provide preliminary evidence that CARD14 expression in ECs may contribute to psoriasis through increased expression of chemokines and facilitating recruitment of immune cells into skin.

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Phosphorylated NF-κB (pNF-κB) was upregulated in psoriatic skin and dermal pNF-κB co-localized with CARD14+ ECs.(a) Immunohistochemistry of pNF-κB in normal, non-lesional, and lesional frozen skin sections. (b) Two-color immunofluorescence of CD31 (green) and pNF-κB (red) in normal, non-lesional, and lesional skin. Representative images; bar = 10 µm. (c) Triple-color immunofluorescence staining of CD31 (green), CARD14 (red), and pNF-κB (blue) in lesional skin. Representative images at 63X magnification; enlarged images are shown to the left.
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pone-0111255-g002: Phosphorylated NF-κB (pNF-κB) was upregulated in psoriatic skin and dermal pNF-κB co-localized with CARD14+ ECs.(a) Immunohistochemistry of pNF-κB in normal, non-lesional, and lesional frozen skin sections. (b) Two-color immunofluorescence of CD31 (green) and pNF-κB (red) in normal, non-lesional, and lesional skin. Representative images; bar = 10 µm. (c) Triple-color immunofluorescence staining of CD31 (green), CARD14 (red), and pNF-κB (blue) in lesional skin. Representative images at 63X magnification; enlarged images are shown to the left.

Mentions: Active NF-κB signaling has been shown previously in psoriatic lesional skin by staining for pNF-κB [12], [13]. These studies focused primarily on pNF-κB in keratinocytes, showing significant staining of the epidermis in LS skin. However, dermal staining for pNF-κB was also observed. To confirm these findings, we performed immunohistochemistry for pNF-κB on normal, NL, and LS skin (Figure 2a). As previously published, normal skin had minimal pNF-κB, NL skin contained moderate amounts of pNF-κB, and LS skin contained the maximal amount of activated NF-κB. Dermal staining was most pronounced in LS skin. To determine if any of the dermal pNF-κB was within endothelial cells, we performed double immunofluorescence studies. Minimal expression of dermal pNF-κB and therefore minimal co-localization with CD31+ ECs was found in normal skin (Figure 2a–b). However, pNF-κB co-localized with CD31+ ECs in the dermis of NL and LS skin, showing that ECs exhibit an activated phenotype in psoriasis (Figure 2b). Triple immunofluorescence studies confirmed that the CARD14+ CD31+ dermal ECs in LS skin contained pNF-κB (Figure 2c). Expression of pNFkB in CD31+ dermal ECs was also confirmed using confocal microscopy (data not shown). In summary, these studies show that CD31+ECs contain the NF-κB scaffolding molecule, CARD14, and have phosphorylated NF-κB in LS and NL skin.


CARD14 expression in dermal endothelial cells in psoriasis.

Harden JL, Lewis SM, Pierson KC, Suárez-Fariñas M, Lentini T, Ortenzio FS, Zaba LC, Goldbach-Mansky R, Bowcock AM, Lowes MA - PLoS ONE (2014)

Phosphorylated NF-κB (pNF-κB) was upregulated in psoriatic skin and dermal pNF-κB co-localized with CARD14+ ECs.(a) Immunohistochemistry of pNF-κB in normal, non-lesional, and lesional frozen skin sections. (b) Two-color immunofluorescence of CD31 (green) and pNF-κB (red) in normal, non-lesional, and lesional skin. Representative images; bar = 10 µm. (c) Triple-color immunofluorescence staining of CD31 (green), CARD14 (red), and pNF-κB (blue) in lesional skin. Representative images at 63X magnification; enlarged images are shown to the left.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4219711&req=5

pone-0111255-g002: Phosphorylated NF-κB (pNF-κB) was upregulated in psoriatic skin and dermal pNF-κB co-localized with CARD14+ ECs.(a) Immunohistochemistry of pNF-κB in normal, non-lesional, and lesional frozen skin sections. (b) Two-color immunofluorescence of CD31 (green) and pNF-κB (red) in normal, non-lesional, and lesional skin. Representative images; bar = 10 µm. (c) Triple-color immunofluorescence staining of CD31 (green), CARD14 (red), and pNF-κB (blue) in lesional skin. Representative images at 63X magnification; enlarged images are shown to the left.
Mentions: Active NF-κB signaling has been shown previously in psoriatic lesional skin by staining for pNF-κB [12], [13]. These studies focused primarily on pNF-κB in keratinocytes, showing significant staining of the epidermis in LS skin. However, dermal staining for pNF-κB was also observed. To confirm these findings, we performed immunohistochemistry for pNF-κB on normal, NL, and LS skin (Figure 2a). As previously published, normal skin had minimal pNF-κB, NL skin contained moderate amounts of pNF-κB, and LS skin contained the maximal amount of activated NF-κB. Dermal staining was most pronounced in LS skin. To determine if any of the dermal pNF-κB was within endothelial cells, we performed double immunofluorescence studies. Minimal expression of dermal pNF-κB and therefore minimal co-localization with CD31+ ECs was found in normal skin (Figure 2a–b). However, pNF-κB co-localized with CD31+ ECs in the dermis of NL and LS skin, showing that ECs exhibit an activated phenotype in psoriasis (Figure 2b). Triple immunofluorescence studies confirmed that the CARD14+ CD31+ dermal ECs in LS skin contained pNF-κB (Figure 2c). Expression of pNFkB in CD31+ dermal ECs was also confirmed using confocal microscopy (data not shown). In summary, these studies show that CD31+ECs contain the NF-κB scaffolding molecule, CARD14, and have phosphorylated NF-κB in LS and NL skin.

Bottom Line: CARD14 is a scaffolding protein that regulates NF-κB activation, and psoriasis-associated CARD14 mutations lead to enhanced NF-κB signaling.CARD14 is expressed mainly in epidermal keratinocytes, but also in unidentified dermal cells.Transfection of dermal ECs with psoriasis-associated CARD14 mutations resulted in increased expression of several chemokines, including CXCL10, IL-8, and CCL2.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Investigative Dermatology, The Rockefeller University, New York, New York, United States of America.

ABSTRACT
Mutations in the caspase recruitment domain, family member 14 (CARD14) gene have recently been described in psoriasis patients, and explain the psoriasis susceptibility locus 2 (PSORS2). CARD14 is a scaffolding protein that regulates NF-κB activation, and psoriasis-associated CARD14 mutations lead to enhanced NF-κB signaling. CARD14 is expressed mainly in epidermal keratinocytes, but also in unidentified dermal cells. In this manuscript, the identity of the dermal cell types expressing CARD14, as well the potential functional consequence of overactive CARD14 in these dermal cell types, was determined. Using two-color immunofluorescence, dermal CARD14 did not co-localize with T-cells, dendritic cells, or macrophages. However, dermal CARD14 did highly co-localize with CD31(+) endothelial cells (ECs). CARD14 was also expressed non-dermal endothelial cells, such as aortic endothelial cells, which may indicate a role of CARD14(+)ECs in the systemic inflammation and cardiovascular comorbidities associated with psoriasis. Additionally, phosphorylated NF-κB was found in psoriatic CARD14(+) CD31(+) ECs, demonstrating this pathway is active in dermal ECs in psoriasis. Transfection of dermal ECs with psoriasis-associated CARD14 mutations resulted in increased expression of several chemokines, including CXCL10, IL-8, and CCL2. These results provide preliminary evidence that CARD14 expression in ECs may contribute to psoriasis through increased expression of chemokines and facilitating recruitment of immune cells into skin.

Show MeSH
Related in: MedlinePlus