Fig 1.
Characteristics of IMQ induced skin inflammation in wild-type mice.
Topical application of IMQ results in increased epidermal thickness, infiltration of various immune cells and release of several pro-inflammatory cytokines and chemokines relevant for psoriasis. The figure has been created with BioRender.com and the read-outs are mainly from published data (cellular infiltration, cytokines and chemokines), but un-published in-house data (epidermal thickness pictures) are also included.
Fig 2.
The group denoted IMQ alt. (3x) initiated IMQ application the day before the final boost (day 6) and was dosed with IMQ on day 6, 7 and 8. The groups denoted IMQ (2x) and IMQ (3x) initiated IMQ application on the day after the final boost (day 8). Group IMQ (2x) was dosed with IMQ on day 8 and 9 and group IMQ (3x) was dosed with IMQ on day 8, 9 and 10. Boosted mice with a clipped back area were used as controls (denoted Naïve (2x) and (3x) indicating that the mice were either terminated along with the mice receiving two or three IMQ applications).
Fig 3.
IMQ induced a clinical skin inflammation and increased epidermal thickness.
(a) Cumulative clinical score of the skin before the first, second and third IMQ application (Denoted pre 1., 2. and 3. IMQ in the figure). (b) Epidermal thickness measured with Visiopharm software from Masson’s Trichrome (MT) stained skin at sacrifice. (c) Masson’s Trichrome (MT) stained skin. Representative slides are shown. The bar equals 250 μm.
Fig 4.
Immunohistochemical stainings to evaluate the presence of human and murine immune cells in the skin.
IHC stainings of skin with (a) anti-KU80, (b) anti-CD3, (c) anti-CD68, (d) anti-DC-lamp, (e) anti-MPO. Representative slides are shown. The bar equals 100 μm. A higher magnification of areas with positive staining has been included in slides with positive cells. In slides with no positive staining higher magnifications have not been shown.
Fig 5.
Quantification of human and murine immune cells in the skin.
Human and murine leukocytes were identified in the skin by IHC staining. Quantification was performed by measuring the positive staining area of (a) all human cells (KU80), (b) human T-cells (CD3), (c) human monocytes/macrophages (CD68), (d) activated human DCs (DC-lamp) and (e) murine/human neutrophils (MPO) per skin biopsy length with Visiopharm software.
Fig 6.
Human and murine protein biomarkers in skin lysates from hFlt3L boosted BRGSF-HIS mice.
Human IL-1β (a), IL-6 (b), TNF-α (c) and murine IL-1β (d), IL-6 (e), TNF-α (f), CXCL-1 (g) and MIP-3α (h) protein levels analysed in skin lysates at terminations by the MSD platform. LLOD identifies the lower limit of detection.
Fig 7.
Serum levels of murine protein biomarkers in hFlt3L boosted BRGSF-HIS mice.
Murine TNF-α (a) and IL-6 (b) protein levels analysed in serum at terminations by the MSD platform. LLOD identifies the lower limit of detection.
Fig 8.
Human immune cells in blood and spleen.
(a) Percent and (b) number of human γδTCR+ of human CD45+ cells and (c) percent and (d) number of human pDCs of human CD45+ cells were analyzed in spleen cell suspensions by flow cytometry at terminations. Percent and (e) number (f) of human monocytes of human CD45+ cells were analyzed by flow cytometry on blood at terminations.
Fig 9.
Murine granulocytes in spleen cell suspensions.
(a) Percent total murine granulocytes of murine CD45+ cells (Percentage calculated as: (#muLy6G+ + #muLy6Gint) / #muCD45+ cells)). Percent mature (b) and immature (c) murine granulocytes of total granulocytes analyzed by flow cytometry at terminations. One mouse was excluded from IMQ alt. (3x) group due to shifting of the Ly6G population which made it difficult to distinguish between the two sub-populations.