Bacterially Delivered miRNA-Mediated Toll-like Receptor 8 Gene Silencing for Combined Therapy in a Murine Model of Atopic Dermatitis: Therapeutic Effect of miRTLR8 in AD

In atopic dermatitis (AD), skin inflammation is caused by complex interactions between genetic disposition and aberrant innate/adaptive immune responses. Toll-like receptors (TLRs) are key molecules in the innate/adaptive immune response as they recognize various molecular motifs associated with pathogens. Among them, TLR8 is implicated in eczematous skin reactions. We investigated the combined therapeutic effects of TLR8 gene silencing by the bacterial delivery of miRNA. We used Salmonella as a vector to deliver TLR8 miRNA. The recombinant strain of Salmonella enterica subsp. enterica serovar Typhimurium (ST) expressing TLR8 miRNA (ST-miRTLR8) was prepared for knockdown of TLR8. After oral administration of ST-miRTLR8 into mice, we observed the cytokine levels, skin pathology and scratching behaviors in an AD-like mouse model. TLR8 down-regulation decreased macrophage-derived chemokine concentrations in activated human mast cells. Serum IgE and interleukin-4 production were suppressed whereas IFN-γ was induced after oral administration of ST-miRTLR8. Scratching behaviors and skin inflammation were also improved. In addition, attenuated S. typhimurium safely accumulated in mouse macrophages and showed adjuvant effects. This study shows that the recombinant miRNA that expresses the TLR8 miRNA has therapeutic effects by suppressing Th2 inflammation. TLR gene modulation using miRNA via Salmonella vectors will thus have a double-protective effect in the treatment of AD.

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Gomisin M2 Ameliorates Atopic Dermatitis-like Skin Lesions via Inhibition of STAT1 and NF-κB Activation in 2,4-Dinitrochlorobenzene/Dermatophagoides farinae Extract-Induced BALB/c Mice

Molecules ◽

10.3390/molecules26154409 ◽

2021 ◽

Vol 26 (15) ◽

pp. 4409

Author(s):

Jinjoo Kang ◽

Soyoung Lee ◽

Namkyung Kim ◽

Hima Dhakal ◽

Taeg-Kyu Kwon ◽

...

Keyword(s):

Atopic Dermatitis ◽

Oral Administration ◽

Skin Diseases ◽

Nuclear Translocation ◽

Skin Lesions ◽

Biologically Active ◽

Therapeutic Effects ◽

Dermatophagoides Farinae ◽

Tnf Α ◽

Ifn Γ

The extracts of Schisandra chinensis (Turcz.) Baill. (Schisandraceae) have various therapeutic effects, including inflammation and allergy. In this study, gomisin M2 (GM2) was isolated from S. chinensis and its beneficial effects were assessed against atopic dermatitis (AD). We evaluated the therapeutic effects of GM2 on 2,4-dinitrochlorobenzene (DNCB) and Dermatophagoides farinae extract (DFE)-induced AD-like skin lesions with BALB/c mice ears and within the tumor necrosis factor (TNF)-α and interferon (IFN)-γ-stimulated keratinocytes. The oral administration of GM2 resulted in reduced epidermal and dermal thickness, infiltration of tissue eosinophils, mast cells, and helper T cells in AD-like lesions. GM2 suppressed the expression of IL-1β, IL-4, IL-5, IL-6, IL-12a, and TSLP in ear tissue and the expression of IFN-γ, IL-4, and IL-17A in auricular lymph nodes. GM2 also inhibited STAT1 and NF-κB phosphorylation in DNCB/DFE-induced AD-like lesions. The oral administration of GM2 reduced levels of IgE (DFE-specific and total) and IgG2a in the mice sera, as well as protein levels of IL-4, IL-6, and TSLP in ear tissues. In TNF-α/IFN-γ-stimulated keratinocytes, GM2 significantly inhibited IL-1β, IL-6, CXCL8, and CCL22 through the suppression of STAT1 phosphorylation and the nuclear translocation of NF-κB. Taken together, these results indicate that GM2 is a biologically active compound that exhibits inhibitory effects on skin inflammation and suggests that GM2 might serve as a remedy in inflammatory skin diseases, specifically on AD.

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Regulation of Skin Barrier Function via Competition between AHR Axis versus IL-13/IL-4‒JAK‒STAT6/STAT3 Axis: Pathogenic and Therapeutic Implications in Atopic Dermatitis

Journal of Clinical Medicine ◽

10.3390/jcm9113741 ◽

2020 ◽

Vol 9 (11) ◽

pp. 3741

Author(s):

Masutaka Furue

Keyword(s):

Atopic Dermatitis ◽

Barrier Function ◽

Coal Tar ◽

Calcineurin Inhibitors ◽

Skin Barrier ◽

Skin Inflammation ◽

Interleukin 4 ◽

Skin Barrier Function ◽

Barrier Dysfunction ◽

Therapeutic Implications

Atopic dermatitis (AD) is characterized by skin inflammation, barrier dysfunction, and chronic pruritus. As the anti-interleukin-4 (IL-4) receptor α antibody dupilumab improves all three cardinal features of AD, the type 2 cytokines IL-4 and especially IL-13 have been indicated to have pathogenic significance in AD. Accumulating evidence has shown that the skin barrier function is regulated via competition between the aryl hydrocarbon receptor (AHR) axis (up-regulation of barrier) and the IL-13/IL-4‒JAK‒STAT6/STAT3 axis (down-regulation of barrier). This latter axis also induces oxidative stress, which exacerbates inflammation. Conventional and recently developed agents for treating AD such as steroid, calcineurin inhibitors, cyclosporine, dupilumab, and JAK inhibitors inhibit the IL-13/IL-4‒JAK‒STAT6/STAT3 axis, while older remedies such as coal tar and glyteer are antioxidative AHR agonists. In this article, I summarize the pathogenic and therapeutic implications of the IL-13/IL-4‒JAK‒STAT6/STAT3 axis and the AHR axis in AD.

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Quercetin effects on adaptive immune response in experimental periodontitis of bacterial-immune genesis

Pharmacia ◽

10.3897/pharmacia.68.e70883 ◽

2021 ◽

Vol 68 (4) ◽

pp. 877-882

Author(s):

Andrii Demkovych ◽

Yurii Bondarenko ◽

Vitaliy Shcherba ◽

Vitalii Luchynskyi ◽

Volodymyr Vitkovskyy ◽

...

Keyword(s):

Immune Response ◽

Process Development ◽

Adaptive Immune Response ◽

Relative Number ◽

Therapeutic Effects ◽

Percentage Increase ◽

Humoral Immune ◽

Periodontal Tissues ◽

Adaptive Immune ◽

Experimental Periodontitis

In the article was studied the effects of flavonol quercetin on indices of adaptive immune response in experimental animals on the 14th day of the experimental bacterial-immune periodontitis development. Indices of immune protection were determined by the relative number of lymphocytes with CD3+, CD4+, CD8+, CD19+, CD16+ and immunoregulatory index (CD4+ / CD8+) in intact animals and on the 14th day of inflammatory process development in periodontal tissues as well as the therapeutic effects of flavonol quercetin. As a result of the study, characterized changes associated with the activity of both the cell-mediated and humoral-immune response were found, both in the development of experimental periodontitis, and apply of flavonol. In particular, there was an increase in the animal’s blood relative amount of CD8+, CD16+ cells on the 14th day, and content of CD3+, CD4+, CD19+ was decreased. In this case, the immunoregulatory index (CD4+ / CD8+) as an important index of immunological activity was decreased. The apply of flavonol quercetin in the period development of bacterial-immune periodontitis animals functional activity of the T-cell line of the immune system was increased, as evidenced percentage increase of B- and T-cells due to T-helper cells decrease as well as T-killers content during this period of inflammatory reaction in the periodontal complex, in comparison with animals, which were not treated.

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Immune Alterations in IgE and Non IgE-Associated Atopic Dermatitis

The Open Dermatology Journal ◽

10.2174/1874372201408010060 ◽

2014 ◽

Vol 8 (1) ◽

pp. 60-67 ◽

Cited By ~ 1

Author(s):

Giampaolo Ricci ◽

Elisabetta Calamelli ◽

Francesca Cipriani

Keyword(s):

Atopic Dermatitis ◽

Immune System ◽

Complex Disease ◽

Adaptive Immune Response ◽

Adaptive Immune System ◽

Skin Barrier ◽

Underlying Mechanism ◽

Adaptive Immune ◽

New Findings ◽

Immune Alterations

Atopic dermatitis is a complex disease in which a strong interaction between alterations of skin barrier and the adaptive immune system coexists. In the recent years, new findings have underlined the importance of skin proteins, especially filaggrin, which participate to the outmost layers of the skin. To strengthen this physical barrier, many factors are available, such as antimicrobial peptides, chemokines and cytokines produced by keratinocytes. Skin disruption can easily allow the allergen penetration and the local keratinocytes can promote the adaptive immune response toward a Th2 phenotype. On the other side, allergic Th2 cytokines may downregulate the production of skin barrier proteins, facilitating the penetration of allergens. Moreover, data on murine models show the absolute relevance of the systemic immune system to develop clinical skin reaction. Since the clinical aspect of patients with AD does not show different patterns whatever is the prevalent underlying mechanism, in clinical practice it is difficult to translate the different endotypes beside the IgE and non IgE associated forms. The aim of this review is to point out to the most recent knowledge in this field, which makes AD more difficult to frame in a unique clinical entity.

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Interleukin-8 Levels in the Stratum Corneum as a Biomarker for Monitoring Therapeutic Effect in Atopic Dermatitis Patients

International Archives of Allergy and Immunology ◽

10.1159/000512965 ◽

2021 ◽

pp. 1-15

Author(s):

Susumu Murata ◽

Sakae Kaneko ◽

Eishin Morita

Keyword(s):

Atopic Dermatitis ◽

Topical Corticosteroid ◽

Stratum Corneum ◽

Corticosteroid Treatment ◽

Skin Inflammation ◽

Interleukin 8 ◽

Tape Stripping ◽

Therapeutic Effects ◽

Local Skin ◽

Before And After

Introduction: The stratum corneum contains several growth factors and cytokines that are synthesized in keratinocytes. We previously reported that the amount of interleukin-8 in the stratum corneum (scIL-8) is related to the severity of local skin inflammation in atopic dermatitis (AD). However, it is unknown whether scIL-8 levels reflect pharmacologic responses to a therapeutic intervention in AD patients. Therefore, in this study, we aimed to investigate whether the improvement of dermatitis in AD is correlated with scIL-8 levels before and after topical corticosteroid treatment. Methods: Stratum corneum samples were collected from 22 AD patients using the noninvasive tape-stripping method before treatment, 2 weeks after topical treatment, and 4–6 weeks after treatment. Results: scIL-8 levels on the forearm reduced significantly from 790 ± 348 pg/mg before treatment to 163 ± 68 pg/mg 2 weeks after treatment and 100 ± 37 pg/mg 4–6 weeks after corticosteroid treatment. scIL-8 levels on the abdomen also reduced significantly from 902 ± 391 to 142 ± 38 pg/mg at the end of study. The reduction in scIL-8 levels was associated with the improvement in local skin severity in AD. We also found that scIL-8 levels, along with blood biomarker levels (serum thymus and activation-regulated chemokine, lactate dehydrogenase, and %eosinophil), decreased significantly after the treatment. Conclusion: The scIL-8 concentration decreases with improvements in skin symptoms in AD patients after topical corticosteroid treatment; thus, it may be a suitable biomarker for monitoring therapeutic effects in AD patients.

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The lytic activity of VSV-GP treatment dominates the therapeutic effects in a syngeneic model of lung cancer

British Journal of Cancer ◽

10.1038/s41416-019-0574-7 ◽

2019 ◽

Vol 121 (8) ◽

pp. 647-658

Author(s):

Liesa-Marie Schreiber ◽

Carles Urbiola ◽

Krishna Das ◽

Bart Spiesschaert ◽

Janine Kimpel ◽

...

Keyword(s):

Lung Cancer ◽

Adaptive Immune Response ◽

Mouse Lung ◽

Therapeutic Effects ◽

Cancer Model ◽

Adaptive Immune ◽

Syngeneic Mouse ◽

Tumour Spread ◽

Lung Cancer Model

Abstract Background Oncolytic virotherapy is thought to result in direct virus-induced lytic tumour killing and simultaneous activation of innate and tumour-specific adaptive immune responses. Using a chimeric vesicular stomatitis virus variant VSV-GP, we addressed the direct oncolytic effects and the role of anti-tumour immune induction in the syngeneic mouse lung cancer model LLC1. Methods To study a tumour system with limited antiviral effects, we generated interferon receptor-deficient cells (LLC1-IFNAR1−/−). Therapeutic efficacy of VSV-GP was assessed in vivo in syngeneic C57BL/6 and athymic nude mice bearing subcutaneous tumours. VSV-GP treatment effects were analysed using bioluminescent imaging (BLI), immunohistochemistry, ELISpot, flow cytometry, multiplex ELISA and Nanostring® assays. Results Interferon insensitivity correlated with VSV-GP replication and therapeutic outcome. BLI revealed tumour-to-tumour spread of viral progeny in bilateral tumours. Histological and gene expression analysis confirmed widespread and rapid infection and cell killing within the tumour with activation of innate and adaptive immune-response markers. However, treatment outcome was increased in the absence of CD8+ T cells and surviving mice showed little protection from tumour re-challenge, indicating limited therapeutic contribution by the activated immune system. Conclusion These studies present a case for a predominantly lytic treatment effect of VSV-GP in a syngeneic mouse lung cancer model.

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