Application Of Imiquimod Research Articles

Inhibition of JAK1/STAT3 pathway by 2-methoxyestradiol ameliorates psoriatic features in vitro and in an imiquimod-induced psoriasis-like mouse model

Psoriasis is characterized by hyperproliferative keratinocytes, dilated capillaries and leukocyte infiltration. 2-Methoxyestradiol (2-ME) has shown significant inhibition on proliferation, angiogenesis and inflammation. To evaluate the anti-psoriatic potential of 2-ME, psoriasis-like dermatitis was induced by topical application of imiquimod (IMQ) on the dorsal skin of C57BL/6 mice for seven consecutive days, followed by treatment of vehicle or 2-ME ointment from Day 4 on. The psoriasis area and severity index (PASI) was assessed daily. On Day 8, skin histology and spleen index were assessed. The effects of 2-ME on the proliferation, apoptosis, cell cycle, vascular endothelial growth factor A (VEGFA), and Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathways of HaCaT cells stimulated by interleukin-17 (IL-17A) were detected, together with its effect on the proliferation, tube formation and VEGF receptor expression of human umbilical vein endothelial cells (HUVECs). We found that topical 2-ME treatment significantly improved IMQ-induced psoriasis-like dermatitis and decreased the PASI scores, the activation of STAT3 in the skin (P < 0.05), and the spleen index in mice (P < 0.01). In vitro, 2-ME inhibited the proliferation of HaCaT cells by inducing apoptosis and G2/M phase arrest (P < 0.01). Moreover, 2-ME suppressed IL-17A-induced VEGFA (2.5 μM: P < 0.05; 5 μM: P < 0.01) and phosphorylation of STAT3 by blocking p-JAK1 in HaCaT cells and prevented tube formation (P < 0.01) and proliferation by targeting VEGF receptors 1 (VEGFR1) and 2 (VEGFR2) in HUVECs. We conclude that 2-ME alleviated psoriasis in vivo and in vitro by inhibiting JAK1/STAT3 pathway and was a promising therapeutic agent for psoriasis.

Brief Maternal Separation Promotes Resilience to Anxiety-like and Depressive-like Behaviors in Female C57BL/6J Offspring with Imiquimod-Induced Psoriasis.

Background: Psoriasis is a common chronic inflammatory skin disease that often causes depression. Early life experience affects brain development and relates to depression. Whether the effect of different MS protocols in early life on anxiety-like and depressive-like behaviors in female offspring with imiquimod (IMQ)-induced psoriasis is unknown. Methods: C57BL/6J mice were subjected to no separation (NMS), brief MS (15 min/day, MS15) or long MS (180 min/day, MS180) from postpartum days (PPD) 1 to PPD21. Then, 5% imiquimod cream was applied for 8 days in adults. Behavioral tests, skin lesions and hippocampal protein expression were also assessed. Results: We found significant psoriasis-like skin lesions in female mice following IMQ application, and mice showed anxiety-like and depressive-like behaviors. Further, increased microglial activation and decreased expression of neuroplasticity were detected in mice following IMQ application. However, after MS15 in early life, mice showed decreased anxiety-like and depressive-like behaviors, indicating resilience. Further, inhibited hippocampal neuroinflammation and increased neuroplasticity were detected. Conclusions: Collectively, this study confirms that brief MS confers resilience to the behavior deficits in female offspring with IMQ-induced psoriasis and reverses the activation of neuroinflammation and the damage of neuroplasticity injury.

Effects of splenectomy on skin inflammation and psoriasis-like phenotype of imiquimod-treated mice

Imiquimod (IMQ) is widely used as animal model of psoriasis, a chronic inflammatory skin disorder. Although topical application of IMQ to back skin causes splenomegaly in mice, how the spleen affects the psoriasis-like phenotype of IMQ-treated mice remains unclear. In this study, we analyzed the cellular composition of spleen and measured metabolites in blood of IMQ-treated mice. We also investigated whether splenectomy influences the degree of skin inflammation and pathology in IMQ-treated mice. Flow cytometry showed that the numbers of CD11b+Ly6c+ neutrophils, Ter119+ proerythroblasts, B220+ B cells, F4/80+ macrophages, and CD11c+ dendritic cells in the spleen were significantly higher in IMQ-treated mice compared to control mice. An untargeted metabolomics analysis of blood identified 14 metabolites, including taurine and 2,6-dihydroxybenzoic acid, whose levels distinguished the two groups. The composition of cells in the spleen and blood metabolites positively correlated with the weight of the spleen. However, splenectomy did not affect IMQ-induced psoriasis-like phenotypes compared with sham-operated mice, although splenectomy increased the expression of interleukin-17A mRNA in the skin of IMQ-treated mice. These data suggest that the spleen does not play a direct role in the development of psoriasis-like phenotype on skin of IMQ-treated mice, though IMQ causes splenomegaly.

MiRNA-203b-3p Induces Acute and Chronic Pruritus through 5-HTR2B and TRPV4

Growing evidence indicates that transient receptor potential (TRP) channels contribute to different forms of pruritus. However, the endogenous mediators that cause itch through transient receptor potential channels signaling are poorly understood. In this study, we show that genetic deletion or pharmacological antagonism of TRPV4 attenuated itch in a mouse model of psoriasis induced by topical application of imiquimod. Human psoriatic lesions showed increased expression of several microRNAs, including the miR-203b-3p, which induced a calcium ion response in rodent dorsal root ganglion neurons and scratching behavior in mice through 5-HTR2B activation and the protein kinase C‒dependent phosphorylation of TRPV4. Computer simulation revealed that the miR-203b-3p core sequence (GUUAAGAA) that causes 5-HTR2B/TRPV4-dependent itch targets the extracellular side of 5-HTR2B by interacting with a portion of the receptor pocket consistent with its activation. Overall, we reveal the unconventional pathophysiological role of an extracellular microRNA that can behave as an itch promoter through 5-HTR2B and TRPV4.

Imiquimod-induced ex vivo model of psoriatic human skin via interleukin-17A signalling of T cells and Langerhans cells.

Several in vitro models have been developed to study the mechanisms involved in psoriasis and to screen new antipsoriatic drugs. However, most of them use single-cell or reconstructed human skin models that did not have complex anatomy of human skin. Thus, this study aimed to create a new model of psoriasis-like dermatitis using human skin under in vitro conditions. To perform this, human skin explants were topically treated with imiquimod (IMQ) or vehicle for 2, 3 or 6 consecutive days. Some explants were treated with an anti-psoriatic drug or antibody anti-interleukin-17A (IL-17A). Topical application of IMQ increased total epidermal area, epidermal proliferation and keratinocyte differentiation at 3, 4 or 7 days. The protein levels of CD3 were augmented in the IMQ-treated human skin explants at 7 days reflecting the activation of T cells. Topical IMQ promoted a higher protein and mRNA levels of IL-17A in human skin ex vivo. Immunofluorescence analysis showed CD207-positive Langerhans cells (LCs) and CD3-positive T cells expressing IL-17A in IMQ-treated human skin explants at 7 days. In addition, administration of antibody anti-IL-17A or an anti-psoriatic drug inhibited IMQ-induced increase in the epidermal thickness in ex vivo human skin at 7 days. In conclusion, topical IMQ application promotes epidermal changes in ex vivo human skin that resemble to human psoriatic skin lesions. Moreover, IMQ-induced production of IL-17 by LCs and T cells is critical to development of psoriasis-like inflammation in our model. This new model is suitable for in vitro screening of antipsoriatic drugs.

Dimeric translationally controlled tumor protein-binding peptide 2 attenuates imiquimod-induced psoriatic inflammation through induction of regulatory T cells.

Psoriasis is a chronic skin inflammation caused by a dysfunctional immune system, which causes systemic inflammation in various organs and tissues. Due to the risk of systemic inflammation and recurrence of psoriasis, it is important to identify the critical targets in the pathogenesis of psoriasis and develop targeted therapeutics. Dimerized translationally controlled tumor protein (dTCTP) promotes immune cell activation as a pro-inflammatory cytokine and plays a role in developing allergic diseases such as asthma and rhinitis. Here, we sought to explore whether dTCTP and its inhibition contributed to the development and control of imiquimod (IMQ)-induced psoriasis. Topical application of IMQ inflamed the skin of the back and ear, increased inflammatory cytokines, and decreased regulatory T cell markers. Interestingly, TCTP was significantly increased in inflamed skin and immune cells such as T cells, B cells, and macrophages after IMQ treatment and was secreted into the serum to undergo dimerization. Extracellular dTCTP treatment selectively suppressed regulatory T (Treg) cells, not other effector T helper (Th) cells, and increased M1 macrophages. Moreover, dTCTP-binding peptide 2 (dTBP2), a dTCTP inhibitor peptide, effectively attenuated the systemic inflammatory responses, including Th17 cell response, and alleviated psoriatic skin inflammation. dTBP2 blocked dTCTP-mediated Treg suppression and stimulated the expression of Treg cell markers in the spleen and inflammatory skin lesions. These results suggest that dTCTP dysregulated immune balance through Treg suppression in psoriatic inflammation and that functional inhibition of dTCTP by dTBP2 maintained immune homeostasis and attenuated inflammatory skin diseases by expanding Treg cells.

486 Glibenclamide ameliorates skin inflammation in a TRPM4 gain-of-function murine model of imiquimod-mediated psoriasiform dermatitis

A heterozygous (het) gain-of-function (GoF) mutation (I1033M or I1040T) of TRPM4, a calcium-activated nonselective monovalent cation channel, has been reported as the cause of progressive symmetric erythrokeratodermia. This genetic disease is characterized by red, scaly plaques in periorificial and acral regions that are reminiscent of human psoriasis. To determine whether the TRPM4 mutation alone is sufficient to lead to skin disease, we created a mouse (TRPM4I1029M, genetically equivalent to the human I1033M mutation) using CRISPR-Cas9 methods. Unperturbed homo and het TRPM4I1029M mice showed no apparent skin changes compared to wild type (WT) littermates. We compared the response of TRPM4I1029M and WT littermates to topical imiquimod (IMQ) application, a model of psoriasiform dermatitis (PsD). Het TRPM4I1029M showed enhanced skin inflammation with greater clinical severity scores (200% vs WT), thicker epidermis, more infiltration of CCR6+ γδ low T cells, higher expression of p-STAT3 protein, and higher mRNA levels of Il17a (1.5-fold) compared to WT mice. In vivo dendritic cell (DC) migration assay showed enhanced DC migration in het TRPM4I1029M. Cell cycle analysis showed that keratinocytes (KCs) from het TRPM4I1029M were in proliferative cell cycle status compared to KCs from WT mice. However, TRPM4I1029M did not show exaggerated responses in other in vivo models such as croton oil-induced irritant dermatitis and DNFB-induced contact dermatitis models. Glibenclamide, a classic diabetes drug, is a known TRPM4 inhibitor and ameliorated IMQ-PsD both in TRPM4I1029M and WT mice with milder clinical scores (50% vs vehicle), thinner epidermis, less infiltration of CCR6+ γδ low T cells, and lower mRNA levels of Il17a (30% vs vehicle). Our results suggest a possible therapeutic application of TRPM4 inhibitors in psoriasis.

The Role of PD-L1 on Langerhans Cells in the Regulation of Psoriasis

Langerhans cells (LCs) are skin-resident cells with potent antigen-presenting cell capabilities, which reportedly play some roles in the development of psoriasis, an inflammatory skin disease mediated by IL-17A‒producing cells, T helper 17 cells, and TCR-γδlow T cells. LCs in psoriatic skin lesions but not in normal skin express PD-L1, which binds to PD-1, an immune checkpoint molecule, to negatively regulate immune reactions. The aim of this study is to elucidate the regulatory role of LCs through the PD-1/PD-L1 axis in a murine model of imiquimod-induced psoriasis-like dermatitis. Imiquimod application on wild-type C57BL/6J mice induced PD-L1 expression on LCs both in the ear skin and skin-draining lymph nodes. To further identify the functional role of PD-L1 expressed on LCs, we generated conditional knockout mice lacking PD-L1 expression on LCs (Pd-l1-cKO mice). Pd-l1-cKO mice presented significantly more severe imiquimod-induced psoriasis-like dermatitis than their control littermates. Flow cytometric analysis showed that the frequency of activated IL-17A‒producing γδlow T cells was increased in the ear skin samples, and IL-17A production by CCR6+ migrating γδlow T cells increased in the skin-draining lymph nodes in imiquimod-applied Pd-l1-cKO mice than in control littermates. Collectively, LCs disrupt the exacerbation of psoriasis through PD-L1.

Treatment of basal cell carcinoma: An overview

Basal cell carcinoma (BCC) is the most common cutaneous cancer worldwide, but accounts for only 2–4% of skin cancers in Asian population. Tumor characteristics such as size, location, pathology, and risk of recurrence, as well as treatment tolerability, cost, and patient preference influence the selection of treatment. The goal of treatment is complete tumor removal in superficial/locally invasive BCCs and prolongation of survival in the rare setting of metastatic BCC. Various treatment options available include conventional surgical excision, Mohs micrographic surgery, cryosurgery, electrodesiccation and curettage, topical application of imiquimod or 5-fluorouracil, and photodynamic, and radiotherapy. Surgical excision and Mohs surgery are preferred because of low recurrence rate and the possibility to evaluate the clearance through histology. In the treatment of metastatic or locally advanced lesions, hedgehog pathway inhibitors and the recently approved drug cemiplimab can be beneficial. Sun protection and regular skin self-checks are recommended for all patients with BCC. This literature review gives an overview of the treatment of BCC.

Abstract CT245: First in human CD200 activation receptor ligand and tumor lysate vaccine immunotherapy for recurrent glioblastoma in adults

Abstract Background: Immunotherapy has revolutionized clinical management of malignancies rendering cancers that previously had a dismal prognosis more manageable. Unfortunately, these therapies can cause serious immune-related adverse events, including death. In CNS cancers, currently available immunotherapies have limited to no efficacy. Our work in glioblastoma (GBM), which is considered one of the most aggressive cancers as well as being universally fatal, is focused on the CD200 immune checkpoint that modulates the immune system through the inhibitory receptor (CD200R1) and activation receptor (CD200AR). The peptide checkpoint ligand (CD200AR-L) was developed to target the CD200AR that downregulates the inhibitory PD-1/PD-L1, CTLA4 and CD200R immune checkpoints, permitting immune system activation that is less hampered by the inhibitory effects of GBM. Following a positive clinical trial in dogs that spontaneously developed high-grade gliomas, we initiated a first-in-human Phase 1 Study of the CD200 Activation Receptor Ligand (CD200AR-L) and Allogeneic Tumor Lysate Vaccine (GBM6-AD) immunotherapy for Recurrent Glioblastoma (NCT04642937). Methods: This is a single-center, first-in-human, dose-escalation Phase 1 clinical trial utilizing a 3+3 design at three dose levels; 3.75micrograms/kg, 5micrograms/kg, and 7.25micrograms/kg. Adults 18 years and old with recurrent GBM and no pre-exiting immune-related condition plus adequate performance score were eligible. Accrual started in December 2020 at dose level 1 with a second cohort having received treatment on dose level 2. CD200AR-L is administered on days 1 and 2 by suprascapular intradermal injection following the application of topical imiquimod. On day 2, a fixed dose of the allogeneic vaccine is injected intradermally at the same site. The “Induction Phase” consists of this injection series weekly for 4 weeks with monitoring for dose-limiting toxicity through Day28. The “Maintenance Phase” doses the vaccine series monthly for 2 months, then the “Booster Phase” doses every eight weeks for 24 months or until end of treatment. Patients are monitored for adverse events, including immune-mediated intracranial edema and other immune-related complications, and their neurological function is scored per NANO criteria. Correlative studies include immune monitoring (including expression of CD200R1, PD1/PD-L1, and CTLA-4) and MRI imaging assessment per iRANO criteria. Citation Format: Michael R. Olin, Elizabeth C. Neil, Anne Eaton, Shannon Lunn, Christopher L. Moertel. First in human CD200 activation receptor ligand and tumor lysate vaccine immunotherapy for recurrent glioblastoma in adults [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr CT245.

An Emerging Role of Proanthocyanidins on Psoriasis: Evidence from a Psoriasis-Like Mouse Model.

Background Psoriasis is an immune-mediated, chronic inflammatory disease, and genetic, immune, oxidative stress (OS), and environmental factors are all thought to contribute to its occurrence. Proanthocyanidins (PCs) are natural flavonoids consisting of catechins and epicatechins which have anti-inflammatory and anti-OS activities. PCs have been widely used to treat various diseases, but reports regarding psoriasis are rare. Objective To investigate the therapeutic effect and potential mechanisms of action of PCs in a psoriasis-like mouse model. Methods Thirty male BALB/c hairless mice were assigned to six groups (n = 5): normal, model, low-dose PCs, medium-dose PCs, high-dose PCs, and control groups. The final five groups were dorsally exposed to 5% imiquimod (IMQ) cream once a day for 6 consecutive days, while the normal group received no intervention. Following the first day of IMQ application, mice in the PC-treated group were dosed with different amounts of PCs daily by oral gavage for six days, whereas mice in the control group received normal saline in the same way. One week later, skin lesions were evaluated by the severity of scoring system based on psoriasis area and severity index (PASI), and pathological alterations were assessed by hematoxylin-eosin (HE) staining. Indicators of inflammation or OS, such as interleukin- (IL-) 17, IL-23, phosphorylated-phosphatidylinositol 3-kinase (p-PI3K), phosphorylated-signal transducer and activator of transcription 3 (p-STAT3), superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), catalase (CAT), vascular endothelial growth factor (VEGF), inducible nitric oxide synthase (iNOS), reactive oxygen species (ROS), and heme oxygenase-1 (HO-1), were determined by ELISA, RT-PCR, western blot, and immunohistochemistry (IHC) analysis. Results IMQ administration induced the formation of large dark red plaques with thickly layered scales on the dorsal skin of mice; nevertheless, the lesions were substantially alleviated by PC administration. Histopathological alterations were observed in both model and control groups with epidermal hyperkeratosis, granulosa layer thinning, acanthosis, downward extension of rete ridges, dermal papillae expansion, capillary hyperplasia, and infiltration by inflammatory cells around blood vessels. These pathological changes, however, were restored by a range of doses of PCs, high-dose PCs in particular. Different doses of PCs significantly lowered the spleen index, levels of inflammatory or oxidative proteins (IL-17, IL-23, MDA, ROS, p-PI3K, and p-STAT3), and the mRNA expression of Il-17, Il-23, Vegf, and iNos. Protein and mRNA levels of anti-OS and anti-inflammatory biomarkers, including SOD, CAT, GSH, and HO-1, greatly increased after PC treatment, especially at the highest dose. Conclusions Our findings reveal that PCs ameliorate psoriasis-like symptoms, suppressing the inflammatory response and mitigating OS damage in an IMQ-induced psoriasis-like mouse model. These effects are probably related to the inactivation of STAT3 and PI3K and activation of HO-1 signaling.

Systemic TLR7/8 micelles trigger a novel and potent anti-tumor response by strong recruitment of neutrophils leading to massive tumor cell killing.

2576 Background: Clinical use of TLR7/8 agonists is currently restricted to topical application of Imiquimod for treatment of superficial basal cell carcinoma since systemic administration is prevented due to dose-limiting toxicity. Here, we present a novel micelle-based drug delivery technology containing a lipid-anchored TLR7/8 agonist for intravenous administration. The MBS8 formulation shows good efficacy in mouse cancer models, is well tolerated in rodents and non-human primates and is currently being tested in clinical studies (NCT04855435). Methods: MBS8 was tested in 12 syngeneic mouse cancer models as monotherapy or in combination anti-PD-1 and anti-PD-L1 antibodies. MBS8 was administered IV as slow bolus. Tumors and spleens were analysed using histology, immunohistochemistry, ELISPOT and flow cytometry. Phenotyping of tumor microenvironment was done using flow cytometry and Nanostring analyses. Safety and tolerability were tested in mice, rats and monkeys. Safety-related and PD biomarkers were identified in monkeys using a multiplex MSD technology and validated in whole human blood stimulated with MBS8 ex-vivo. Results: In several syngeneic mouse tumor models, MBS8 led to complete eradication of established tumors. The complete responders showed tumor rejection upon re-challenge. In these mice, a CD8+-dependent tumor-specific immune memory response was evident. Tumors demonstrated a massive tissue necrosis ( &gt; 80%) within 24-48h after the first drug administration and showed massive neutrophil infiltration 6h after dosing. Antigen specific CD8+ T-cells were detected in tumors 24 hours after treatment with increased CD8+:Treg cell ratio and enhanced antigen presentation in tdLN. MBS8 showed both additive and synergistic effect when combined with anti-PD-1 or anti-PD-L1 antibody treatment. Moreover, in tumors being resistant to anti-PD-1 treatment, co-administration of MBS8 reverted sensitivity to anti-PD-1 treatment in a synergistic manner. A panel of cytokines induced by MBS8 upon repeat dose administration in monkeys was identified in plasma as potential biomarkers. Ten of them were further evaluated in whole human blood treated with MBS8 ex vivo. Currently, these cytokines are being used in Phase I clinical studies. Conclusions: MBS8 showed significant anti-cancer activity in multiple in vivo solid tumor models when administered either as monotherapy or in combination with ICIs. MBS8 demonstrated a novel mode of action with neutrophils playing a central role as primary effector cells causing a rapid killing of tumors. Further, adaptive immune response was initiated including generation of tumor specific CD8+ cells and establishment of the immune memory. MBS8 was well tolerated in rodents and cynomolgus monkeys at the dose levels above therapeutic effective doses identified in mice, thus providing a good therapeutic window.

Dietary olive oil intake aggravates psoriatic skin inflammation in mice via Nrf2 activation and polyunsaturated fatty acid imbalance

Psoriasis is a chronic inflammatory skin disease, which does not have effective treatment options. However, olive oil has been suggested as an alternative to treat psoriasis, but no study has evaluated the mechanisms involved in the effects of olive oil on psoriasis. Thus, the current study investigated whether olive oil could ameliorate psoriasiform skin inflammation. To test this, mice received topical application of imiquimod to induce inflammation and were treated orally with olive oil. Human immortalized keratinocytes were also treated with imiquimod and olive oil. Epidermal thickness and keratinocyte proliferation were increased in imiquimod-induced lesions of olive-oil-treated animals. In both in vitro and in vivo studies, protein levels of nuclear factor erythroid 2-related factor 2 (Nrf2) were elevated following imiquimod and olive oil administration. Inhibition of Nrf2 abolished the increased proliferation of keratinocytes treated with imiquimod and olive oil, demonstrating the role of Nrf2 in olive oil-mediated exacerbation of psoriasiform skin inflammation. In addition, lower levels of linoleic acid and higher levels of oleic acid were observed in imiquimod- and olive-oil-treated animals, which may also contribute to the adverse effects of olive oil on psoriasis. In conclusion, dietary intake of olive oil aggravates the symptoms of psoriatic skin lesions through the overexpression of Nrf2 and an imbalance in oleic and linoleic acids levels, suggesting that a diet rich in olive oil may have significant negative effects on psoriasis.

Topical Formulation of Noscapine, a Benzylisoquinoline Alkaloid, Ameliorates Imiquimod-Induced Psoriasis-Like Skin Lesions.

Psoriasis is considered an autoimmune inflammatory disease. The disease is spread and diagnosed by the infiltration of inflammatory mediators and cells into the epidermis. Recent theoretical developments have focused on the effectiveness of noscapine (NOS) as a potential alkaloid for being used as a valuable treatment for different diseases. In the present study, psoriasis-like dermatitis was induced on the right ear pinna surface of male Balb/c mice by topical application of imiquimod (IMQ) for ten consecutive days, which was treated with noscapine (0.3, 1, 3, and 10% w/v) or clobetasol (0.05% w/v) as a positive control. The levels of ear length, thickness, severity of skin inflammation, psoriatic itch, psoriasis area severity index (PASI) score, and body weight were measured daily. On the 10th day of study, each ear was investigated for inflammation, fibrosis, proliferation, and apoptosis using histopathological (H&E and Masson's trichrome staining) and immunohistochemistry (Ki67 and p53 staining) assays. Furthermore, the levels of inflammatory biomarkers were characterized by an enzyme-linked immunosorbent assay (ELISA). The results confirmed IMQ-induced psoriasis for five consecutive days. In contrast, noscapine significantly reduced the ear length, thickness, severity of skin inflammation, psoriatic itch and body weight, tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β), interferon-gamma (IFN-γ), interleukin 6 (IL-6), IL-17, and IL-23p19 in a concentration-dependent manner (P < 0.001–0.05 for all cases). Overall, topical noscapine significantly ameliorated both the macroscopical and microscopical features of psoriasis. However, further clinical investigations are required to translate the effects to clinics.

Therapeutic effects of the extract of Sancao Formula, a Chinese herbal compound, on imiquimod-induced psoriasis via cysteine-rich protein 61

ObjectivePsoriasis is a common chronic inflammatory skin disease that is prone to recurrence, and the proinflammatory factor, cysteine-rich protein 61 (Cyr61), is important in its pathophysiology. Long-term clinical practice has shown that Sancao Formula (SC), a Chinese herbal compound, is effective in the treatment of psoriasis, but the precise mechanism remains unknown. In this study, we investigate the mechanism by which SC extract alleviates imiquimod (IMQ)-induced psoriasis. MethodsThe expression of Cyr61 in psoriatic lesions and normal healthy skin was detected using immunohistochemical analysis to investigate the biological role of Cyr61 in models of psoriatic inflammation. A psoriatic mouse model was established by topical application of IMQ, and the effect of topical application of SC extract was evaluated using the psoriasis area and severity index (PASI) score, hematoxylin-eosin staining, and histopathological features of the skin. Next, a HaCaT cell inflammation model was established using interferon-γ (IFN-γ), and the effect of SC extract on the mRNA and protein levels of Cyr61 and intercellular cell adhesion molecule-1 (ICAM-1) was confirmed using Western blot and quantitative real-time polymerase chain reaction analyses. ResultsImmunohistochemical staining showed that the expression of Cyr61 in psoriatic lesions was higher than that in normal skin samples (78.26% vs 41.18%, P < 0.05), and the number of Cyr61-positive cells in psoriatic lesions was also significantly higher than in normal skin (18.66 ± 2.51 vs 4.33 ± 1.52, P < 0.05). Treatment in mice with IMQ-induced psoriasis showed that SC extract could significantly improve the inflammatory phenotype, PASI score (10.875 ± 0.744 vs 3.875 ± 0.582, P < 0.05), and pathological features compared with those in IMQ model group; SC treatment was also associated with decreased levels of Cyr61 and ICAM-1. In the IFN-γ-induced inflammatory cell model, the mRNA and protein levels of Cyr61 and ICAM-1 were upregulated, while the SC extract downregulated the levels of Cyr61 and ICAM-1. ConclusionThe results provide a theoretical basis for the involvement of Cyr61 in the pathogenesis of psoriasis, and suggest that SC should be used to target Cyr61 for the prevention of psoriasis recurrence.

Estimation of recurrence rates with off-label use of 5% imiquimod as an adjuvant therapy after surgery or as a monotherapy in patients with lentigo maligna.

Objectives The aim of this paper is to present the experience of off-label application and possible decrease of recurrence rates in patients with ledigo maligna, when treated only with 5% imiquimod or imiquimod 5% after surgical excision in clinical narrow margins (&lt;5mm) or wider margins (≥5mm). Materials and Methods A multicenter analysis was performed in 7 European centers and n=149 patients were included in three groups a) those with adjuvant application of imiquimod after surgical excision in histopathologically clear margins (narrow &lt;5mm) (group 1a) or (wider ≥5mm) (group1b), b) those with adjuvant application but with histopathologically involved margins(group2) c) those who received monotherapy with imiquimod(group3). In all patients was applied 5% imiquimod cream ,7 days/week, for 6-13 weeks. Results The recurrence rate in group 1b and group 1a were (6.2% to 5.5% respectively). Thus, the application of imiquimod before or after conventional surgery to narrow margins might result in acceptable recurrence rates, possibly comparable to those achieved with surgical techniques involving complete margin assessment, such as Mohs micrographic surgery, but this hypothesis should be assessed by a randomized trial. The recurrence rate in group 2 was 9.1% at a mean follow-up of 34 months while in group3 Imiquimod monotherapy resulted in complete clinical response in 71.4% of treated patients. Conclusions In conclusion by this study is supported surgery as the primary treatment in operable patients with ledigo maligna. In any case the experience by application of Imiquimod in combination with conservative surgery is encouraging.

Different Spatial and Temporal Roles of Monocytes and Monocyte-Derived Cells in the Pathogenesis of an Imiquimod Induced Lupus Model.

Mounting evidence indicates the importance of aberrant Toll-like receptor 7 (TLR7) signaling in the pathogenesis of systemic lupus erythematosus (SLE). However, the mechanism of disease progression remains unclear. An imiquimod (IMQ)-induced lupus model was used to analyze the lupus mechanism related to the aberrant TLR7 signals. C57BL/6 mice and NZB/NZW mice were treated with topical IMQ, and peripheral blood, draining lymph nodes, and kidneys were analyzed focusing on monocytes and monocyte-related cells. Monocytes expressed intermediate to high levels of TLR7, and the long-term application of IMQ increased Ly6Clo monocytes in the peripheral blood and Ly6Clo monocyte-like cells in the lymph nodes and kidneys, whereas Ly6Chi monocyte-like cell numbers were increased in lymph nodes. Ly6Clo monocyte-like cells in the kidneys of IMQ-induced lupus mice were supplied by bone marrow-derived cells as demonstrated using a bone marrow chimera. Ly6Clo monocytes obtained from IMQ-induced lupus mice had upregulated adhesion molecule-related genes, and after adoptive transfer, they showed greater infiltration into the kidneys compared with controls. RNA-seq and post hoc PCR analyses revealed Ly6Clo monocyte-like cells in the kidneys of IMQ-induced lupus mice had upregulated macrophage-related genes compared with peripheral blood Ly6Clo monocytes and downregulated genes compared with kidney macrophages (MF). Ly6Clo monocyte-like cells in the kidneys upregulated Il6 and chemoattracting genes including Ccl5 and Cxcl13. The higher expression of Il6 in Ly6Clo monocyte-like cells compared with MF suggested these cells were more inflammatory than MF. However, MF in IMQ-induced lupus mice were characterized by their high expression of Cxcl13. Genes of proinflammatory cytokines in Ly6Chi and Ly6Clo monocytes were upregulated by stimulation with IMQ but only Ly6Chi monocytes upregulated IFN-α genes upon stimulation with 2′3′-cyclic-GMP-AMP, an agonist of stimulator of interferon genes. Ly6Chi and Ly6Clo monocytes in IMQ-induced lupus mice had different features. Ly6Chi monocytes responded in the lymph nodes of locally stimulated sites and had a higher expression of IFN-α upon stimulation, whereas Ly6Clo monocytes were induced slowly and tended to infiltrate into the kidneys. Infiltrated monocytes in the kidneys likely followed a trajectory through inflammatory monocyte-like cells to MF, which were then involved in the development of nephritis.

Perianal squamous cell carcinoma in a Quarter Horse colt

Squamous cell carcinoma (SCC) is a squamous epithelial neoplasm. They are solitary tumors with indolent expansion, are locally invasive, and can be seen in any part of the body. The most commonly affected sites are the head, orbital region, and genitalia of horses. Solar irradiation is one of the main causes, especially in depigmented and hairless areas. SCC is the neoplasm most frequently described in equine species. Several treatments have been described in horses, usually associated with surgical excision with different adjuvant therapies, hoping for a better prognosis and a decrease in cancer-related morbidity. Since the recurrence rates are high, especially in regions that do not have an adequate safety margin after tumor removal, multimodality treatment is essential. The aim of this report was to describe a case of perianal SCC in a Quarter Horse colt clinically treated with the immunomodulatory drug imiquimod. A three-year-old, grey-haired, male Quarter Horse was examined for diagnosis and treatment of an ulcerative lesion in the perianal region. According to the owner's report, the lesion appeared approximately 16 months ago. Based on the medical history and clinical examination, SCC was suspected. To confirm the clinical diagnosis, a biopsy was performed. Histological examination showed infiltrative neoplastic proliferation from the epidermis, consisting of malignant keratinocytes associated with hyperkeratosis, neutrophilic infiltrate, and foci of ulceration. The underlying dermis exhibited an intense chronic inflammatory infiltrate. The findings were compatible with SCC. Due to the location and extension of the lesion, we decided to use immunomodulatory therapy with imiquimod. After 16 weeks of imiquimod application, the lesion completely regressed. The horse was clinically followed for two years, and no recurrence was identified.

Lidocaine Ameliorates Psoriasis by Obstructing Pathogenic CGRP Signaling‒Mediated Sensory Neuron‒Dendritic Cell Communication

Psoriasis is a chronic immune-mediated skin disorder with the nervous system contributing to its pathology. The neurogenic mediators of psoriasis are elusive, and whether the intervention of the cutaneous nervous system can treat psoriasis remains to be determined. In this study, we conducted a pilot study using an epidural injection of lidocaine to treat patients with psoriasis. Lidocaine treatment markedly reduced patients' clinical scores and improved an imiquimod-induced rat model of psoriasis as competent as systemic delivery of a TNF-α antibody. Imiquimod application elicited aberrant cutaneous nerve outgrowth and excessive generation of neuropeptide calcitonin gene-related peptide from dorsal root ganglion neurons, both of which were inhibited by epidural lidocaine treatment. Single-cell RNA sequencing unveiled the overrepresentation of calcitonin gene-related peptide receptors in dermal dendritic cell populations of patients with psoriasis. Through disturbing calcitonin gene-related peptide signaling, lidocaine inhibited IL-23 production by dendritic cells cocultured with dorsal root ganglion neurons. Thus, epidural nerve block with lidocaine demonstrates an effective therapy for psoriasis, which suppresses both inordinate sensory nerve growth in the inflamed skin and calcitonin gene-related peptide-mediated IL-23 production from psoriatic dendritic cells.

Prenatal infection predisposes offspring to enhanced susceptibility to imiquimod-mediated psoriasiform dermatitis in mice

Background: Infection can upregulate T helper 17 (Th17) signaling and exacerbate psoriasis. Literature has indicated that prenatal infection induces embryonic development toward Th17 signaling and is a risk factor for developing certain Th17-skewing disorders such as autism in offspring. However, it remains unclear if a prenatal infection is a risk factor for developing psoriasis in offspring. Objectives: We investigated if a prenatal infection predisposes mice offspring to enhanced susceptibility to psoriatic inflammation. Methods: Pregnant C57BL/6 mice were intraperitoneally injected with lipopolysaccharide (LPS) or polyinosinic:polycytidylic acid poly I:C to simulate prenatal bacterial and viral infection, respectively. Phosphate-buffered saline (PBS) injection was used as the control. When mice offspring were at the age of 9–10 weeks, imiquimod (IMQ) cream or control vehicle was applied topically on ears for 5 consecutive days to induce psoriasiform dermatitis. The extent of epidermal hyperplasia and the number of Munro microabscesses were examined by histologic analysis. Protein expressions of neutrophil marker Ly6 g and Th17-associated cytokines were measured by western blotting. Results: Prenatal infection with LPS or poly I:C induced a greater extent of epidermal hyperplasia and more Munro microabscesses after IMQ application as compared with prenatal PBS injection. Mice offspring with prenatal infection also had higher protein expressions of Ly6 g, IL-17a, and interleukin-23 after IMQ stimulation compared with their PBS controls. Conclusion: Prenatal infection predisposes mice offspring to enhanced susceptibility to IMQ-mediated psoriasiform dermatitis. The data obtained from the present animal study suggest that prenatal infection might be a risk factor for developing psoriasis in offspring.