Change In Ear Thickness Research Articles

Cross-reactivity between thiuram disulfides and dithiocarbamates. A study of TETD and ZDEC using mouse models.

Rubber accelerators are used in the vulcanization of rubber. However, rubber accelerators for example tetraethylthiuram disulfide (TETD) and zinc diethyldithiocarbamate (ZDEC) may cause contact allergy. Concomitant reactions between ZDEC and TETD have been observed in patients which could be explained by co- or cross-reactivity. To investigate cross-reactivity between TETD and ZDEC and vice versa. Groups of mice were sensitized with TETD or ZDEC based on reported EC3-values. Proliferation of lymphocytes were measured on day 5. To test cross-reactivity, mice were sensitized and challenged 3 weeks later with TETD or ZDEC. The inflammatory response was measured by changes in ear thickness and the proliferative response in CD4+ and CD8+ T cells in the submandibular and cervical draining lymph nodes. Sensitization of mice with doses of ZDEC 3%, TETD 5.6% or TETD 16.2% induced significant increased ear thickness and proliferation of CD4+ and CD8+ T cells. Challenge with ZDEC or TETD in these groups induced significant increased ear thickness. Challenge with ZDEC in mice sensitized to TETD 5.6% or TETD 16.2% induced significant increased proliferation of CD4+ and CD8+ T cells. We show cross-reactivity between TETD and ZDEC. Patients sensitized to TETD or ZDEC should avoid exposure to both ZDEC and TETD.

Topical treatment of tyrosine kinase 2 inhibitor through borneol-embedded hydrogel: Evaluation for preventive, therapeutic, and Recurrent management of psoriasis

Psoriasis, an immune-mediated inflammatory skin disorder characterized by a chronically relapsing-remitting course, continues to be primarily managed through topical therapy. While oral administration of tyrosine kinase 2 inhibitors (TYK2i) stands as an effective approach for psoriasis treatment, the potential efficacy of topical application of TYK2i remains unexplored. Herein, the carbomer/alginic acid hydrogel is embedded with borneol (BO) as a new topical carrier of TYK2i for achieving enhanced transdermal permeation and anti-psoriasis efficacy. The hydrogel system, i.e., TYK2i-BO-gel, exhibits significantly improved preventative and therapeutic effects in mice models of psoriasiform dermatitis, as evidenced by phenotypical images, psoriasis severity score index (PSI), histology, immunohistochemical staining, and PCR analysis. Remarkably, TYK2i-BO-gel outperforms conventional topical corticosteroid therapy by significantly preventing psoriatic lesion recurrence as measured by a nearly 50 % reduction in ear thickness changes (p < 0.0001), PSI (p < 0.0001) and epidermal thickness (p < 0.05). Moreover, a strengthened anti-inflammatory effect caused by TYK2i-BO-gel is seen in a human skin explant model, implying its potential application for human patients. With the addition of BO, the TYK2i-BO-gel not only increases skin permeability but also inhibits the expression of antimicrobial peptides in keratinocytes and facilitates the anti-Th17 response of TYK2i with suppressed activation of STAT3. Therefore, this work represents the accessibility and effectiveness of TYK2i-BO-hydrogel as a new topical formulation for anti-psoriasis management and shows great potential for clinical application.

Auriculotherapy Modulates Macrophage Polarization to Reduce Inflammatory Response in a Rat Model of Acne.

The inflammatory response is an important part of the pathogenesis of acne vulgaris. Auriculotherapy has been shown to have a good therapeutic effect on this disease. The aim of this study was to explore the mechanism underlying the anti-inflammatory effect of auriculotherapy in the treatment of acne vulgaris. Propionibacterium acnes was injected subcutaneously into the ears of rats to establish an animal model of acne. The auriculotherapy intervention in rats consisted of auricular bloodletting therapy (ABT), auricular point sticking (APS), or a combination of both (ABPS). The anti-inflammatory effects of auriculotherapy were evaluated by measuring changes in ear thickness, local body surface microcirculation in the ear, and serum inflammatory factors in rats. The polarization of macrophages was analyzed by flow cytometry, and the expression of TLR2/NF-κB signaling pathway in the target tissues was analyzed using western blot. ABT, APS, and ABPS all reduced the erythema of ear acne, decreased microcirculation in localized ear acne, and decreased serum levels of TNF-α and IL-1β in rats. Meanwhile, the three interventions reduced M1-type macrophages and increased M2-type macrophages; only APS could reduce the expression of TLR2/NF-κB signaling pathway. ABT, APS, and ABPS can improve the inflammatory symptoms of acne and reduce inflammatory cytokines. APS may exert anti-inflammatory effects by altering macrophage polarization and decreasing TLR2/NF-κB expression.

Cubosomal Betamethasone-Salicylic Acid Nano Drug Delivery System for Enhanced Management of Scalp Psoriasis.

IntroductionBetamethasone dipropionate (BD), a potent corticosteroid, and salicylic acid (SA), a keratolytic agent, have been used in combination to treat scalp psoriasis; however, undesirable side effects associated with their prolonged topical use are inevitable. In this study, BD and SA were loaded into cubosomes, a nanoparticulate system with outstanding biocompatibility, bio-adhesivity and penetration power.MethodsDesign of experiments (DOE) was utilized to prepare thirteen different cubosomal dispersions by emulsification technique using glycerol monoolein (GMO) as a lipid phase and Poloxamer 407 (P407) as a surfactant, sodium carboxymethyl cellulose (SCMC) was added to enhance the dispersions’ rheological properties. The thirteen dispersions were in-vitro characterized for their particle size, polydispersity index (PDI), zeta potential, BD and SA content and rheological behaviour. The desirability of an optimized formula (OF) was set to the smallest particle size, lowest zeta-potential and highest viscosity. The OF was in-vitro characterized for the same parameters in addition to transmission electron microscope imaging and in-vitro drug release. The OF’s anti-psoriatic activity was evaluated in-vivo using an imiquimod-induced psoriasis model.ResultsThe OF achieved a particle size of 197.4 ± 9.47 nm, a PDI of 0.443 ± 0.025, a zeta potential of −44.4 ± 0.141mv, BD content of 105.85 ± 2.290%, SA content of 88.855 ± 2.920% with shear-thinning rheological behaviour and completed in-vitro drug release within 2–3 hours. The in-vivo studies confirmed the cubosomes’ higher anti-psoriatic efficacy over the commercial product with lower changes in ear thickness, spleen to body weight ratio, psoriasis area severity index score and improved histopathological findings.ConclusionThe developed BD SA-loaded cubosomes exhibit promising anti-psoriatic activity attributed to its nano-size and unique lipid content, with enhanced skin penetration and modified rheological properties; increasing the formulation’s in-contact duration with the scalp resulting in lower application frequency and thus reduced BD and SA associated side effects.

Differential Involvement of Programmed Cell Death Ligands in Skin Immune Responses

PD-1 is an immunoregulatory receptor that can bind PD-L1 or PD-L2 expressed on stimulated antigen-presenting cells. In this study, isolated antigen-presenting cells (macrophages and dendritic cells) were cultured with IFN-γ, IL-4, or IL-17A, and the expression of PD-L1 and PD-L2 was compared by flow cytometry. Strong upregulation of PD-L1 expression was observed on IFN-γ stimulation of both antigen-presenting cells as well as in response to IL-17A stimulation of macrophages compared with the expression in unstimulated controls. In contrast, only stimulation with IL-4 could upregulate PD-L2 expression on both antigen-presenting cells. Therefore, experiments were performed in murine models, including DNFB-induced contact hypersensitivity, calcipotriol-induced atopic dermatitis-like skin inflammation, and imiquimod-induced psoriasis-like dermatitis models, to trigger IFN-γ‒mediated T helper type (Th)1-, IL-4‒mediated Th2-, and IL-17A‒mediated Th17-type responses, respectively. In both Th1- and Th17-type immunity models, changes in ear thickness were more severe in Pd-l1‒deficient mice than in wild-type or Pd-l2‒deficient mice. In the Th2-type immunity model, changes in thickness in Pd-l2‒deficient mice were more severe than that in wild-type or Pd-l1‒deficient mice. Collectively, PD-L1 has predominant roles in Th1 and Th17 type immunity, whereas PD-L2 is involved in Th2-type immunity.

CD8+ tissue-resident memory T cells recruit neutrophils that are essential for flare-ups in contact dermatitis.

Allergic contact dermatitis (ACD) is classically described as a delayed-type hypersensitivity reaction. However, patients often experience flare-ups characterized by itching erythema, edema, and often vesicles occurring within hours after re-exposure of previously sensitized skin to the specific contact allergen. Recent studies have indicated that skin-resident memory T (TRM ) cells play a central role in ACD. However, the pathogenic role of TRM cells in allergen-induced flare-ups is not known. By the use of various mouse models and cell depletion protocols, we investigated the role of epidermal TRM cells in flare-up reactions to the experimental contact allergen 1-fluoro-2,4-dinitrobenzene. The inflammatory response was measured by changes in ear thickness, and the cellular composition in epidermis was determined by flow cytometry and confocal microscopy. Finally, adaptive transfer and inhibitors were used to determine the role of TRM cells, neutrophils, and CXCL1/CXCL2 in the response. We show that CD8+ TRM cells initiate massive infiltration of neutrophils in the epidermis within 12h after re-exposure to the contact allergen. Depletion of neutrophils before re-exposure to the allergen abrogated the flare-up reactions. Furthermore, we demonstrate that CD8+ TRM cells mediate neutrophil recruitment by inducing CXCL1 and CXCL2 production in the skin, and that blockage of the C-X-C chemokine receptor type 1 and 2 inhibits flare-up reactions and neutrophil infiltration. As the first, we show that epidermal CD8+ TRM cells cause ACD flare-ups by rapid recruitment of neutrophils to the epidermis.

Inhibitory Effect of Sulfated Polysaccharide from Codium edule P.C. Silva Against 2,4-Dinitrofluorobenzene (DNFB)- Induced Allergic Contact Dermatitis on Female BALB/c Mice.

Purpose: Sulfated polysaccharide from Codium species has been reported for its antiinflammatoryactivities. However, the effect of sulfated polysaccharide from Codium edule on allergic responses has not been studied. The study was conducted to determine the effect ofsulfated polysaccharide (F1) from C. edule on allergic contact dermatitis (ACD) induced by2,4-dinitrofluorobenzene (DNFB) in female BALB/c mice. Methods: F1 was isolated using DEAE sepharose gel chromatography and chemically identifiedby LC-MS analyses. The effects of F1 on changes in ear thickness, allergic responses, andhistology were evaluated. The effects of F1 on the production of inflammatory cytokinesinterferon gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-ɑ) in serum were also quantifiedand compared with standard prednisolone therapy. Results: F1 was identified as a heteropolysaccharide with β-D-galactans and β-L-arabinans units.F1 was non-toxic at 2000 mg/kg. Administration of F1 in DNFB-challenged mice significantlysuppressed the increase in ear thickness, erythema, desquamation, and proliferation ofinflammatory cells. F1 significantly decreased the production of inflammatory markers, IFN-γand TNF-α in a dose-dependent manner when compared to the untreated group (P < 0.05). Conclusion: Results suggest that F1 from C. edule is a bioactive sulfated heteropolysaccharidewith anti-inflammatory activity and might be a valuable candidate molecule for the treatmentof allergic diseases such as ACD.

The Berberis aristata Ameliorates oxazolone induced contact dermatitis: in-vivo and in silico evidences

The objective of the present study was to evaluate the effect of Berberis aristata (BA) against oxazolone induced contact dermatitis in Balb/c mice and molecular docking with TLR-1 and TLR-2. Dermatitis was induced in Balb/c mice by sensitizing with topical application of 100 μl oxazolone (2%) and the effect of BA was screened in two doses (200 mg/kg and 400 mg/kg P.O.). The effect was evaluated by the change in ear thickness, anti-inflammatory cytokine (TNF-α, IL-6, and IL-1β) and oxidative stress on the sixth day in ear tissue homogenate. The ear skin of all group mice was subjected to histological analysis. This work was further evidenced by the docking of berberine with crystal structure of TLR1-TLR2 heterodimer caused by using the binding of tri-acylated lipopeptide (PDB ID: 2Z7X). In this study, we found that a significant reduction in ear thickness was found in BA (200 mg/kg and 400 mg/kg) as compared to 100 μl oxazolone(2%) treated mice. The reduction level of GSH and SOD found in 100 μl oxazolone (2%) sensitized mice. BA (200 mg/kg and 400 mg/kg) treated animals showed an increase in GSH and SOD levels. A significant reduction in inflammatory cytokines was observed in BA treated mice and indicates anti-inflammatory activity against oxazolone. Histopathological analysis showed minimal infiltration of lymphocytes and moderate harm to skin cells and layer in BA treated mice. Docking studies revealed promising binding interaction of berberine withTLR1-TLR2 heterodimer which can attribute to its anti dermatic effect. This present research shows that BA has a dose-dependent effect in contact dermatitis attenuated by oxazolone.

Anti-psoriatic effect of Lavandula angustifolia essential oil and its major components linalool and linalyl acetate

Ethno-pharmacological relevanceLavender oil (LO) is an aromatic/essential oil extracted from Lavandula angustifolia and traditionally used as an aromatherapy massage oil due to its anti-inflammatory and wound healing property and also for providing the relief in other skin conditions such as psoriasis, dermatitis and eczema. However, LO has not been evaluated scientifically for psoriasis like skin inflammation. Aim of the studyThis study was aimed to investigate the LO and its major components linalool (L) and linalyl acetate (LA) against psoriasis like skin inflammation. Materials and methodsAnti-psoriatic activity was done using Imiquimod (IMQ) induced psoriasis like skin inflammation in BALB/c mice. Assessment of anti-psoriatic effect of LO, L and LA was done on the basis of change in ear thickness, psoriasis area severity index (PASI) scoring at alternative day, CosCam scoring using skin analyzer equipped with SkinSys software, biochemical, immunohistochemical and histological investigations. Level of effectiveness against psoriasis was investigated by percent reduction in PASI scores, CosCam scores and level of Th-1 and Th-17 cell expressing cytokines, as compared to the diseased mice. ResultsTopical application of LO 10% showed 73.67% recovery in PASI and 87% in Th-17 cell-specific cytokines towards normal as compared to disease group. L and LA were identified as the major components of LO and favoured ligands for selected psoriasis targets. At 2% topical dose, L and LA showed 64% and 47.61% recovery in PASI scores, respectively. Both, L and LA showed significant recovery in Th-1 specific TNF-α and IL-1β however, only L showed significant recovery of Th-17 cytokines (IL-17 and IL-22). In contrast to LA (which restored granulosis), L restored epidermal hyperplasia and parakeratosis toward the normal condition. On the other hand, L also reduced the expression of NF-κβ, ccr6 and IL-17, while LA reduced the expression of NF-κβ only. At 10% topical dose, LO was observed to be slight irritant while at 2% topical dose, L and LA were found non-irritant to the skin. ConclusionThis study proves the effectiveness of LO and its major phytoconstituents linalool and linalyl acetate against IMQ induced psoriasis like skin inflammation and provides the scientific evidence for topical use of lavender oil.

Hydrogel thickened microemulsion; a local cargo for the co- delivery of cinnamaldehyde and berberine to treat acne vulgaris

For the existence in an advancing world of antibiotics, bacteria struggle and make necessary changes at genetic level. This study was designed to deliver a combination of cinnamaldehyde (CA) and berberine chloride (BER) for the treatment of Propionibacterium acnes induced acne vulgaris. CA and BER were first loaded in a clove oil-based microemulsion and then the resultant microemulsion was converted into a hydrogel system using Carbopol 940. Selection of an appropriate concentration of Carbopol 940 was done based on the visual assessment and viscosity of developed formulations. Different concentration of CA and BER was ensured in CA-BER-ME-Gel and labelled them as F-1, F-2, and F-3. F-1 contained 0.5 %w/w CA and 0.108 %w/w BER. F-2 contained 0.75 %w/w CA and 0.162 %w/w BER. F-3 was composed of 1 %w/w CA and 0.217 %w/w BER. The % cumulative amount of drug permeated from F-1, F-2, and F-3 were found to be 62.16 ± 3.06%, 64.9 ± 2.85% and 68.34 ± 4% respectively after 8 h. The CA and BER loaded suspension gel showed significantly lesser permeation profile (27.14 ± 2.42%) as compared to F-1, F-2 and F-3. The anti-acne potential of F-1, F-2, F-3 and Standard (1% w/w Clindamycin Phosphate gel) in terms of % change in ear thickness was found to be 115 ± 7.27%, 55.18 ± 12.76%, 41.85 ± 8.83% and 30.18 ± 13.94% respectively. These values were significantly better (p < 0.0001) than the disease control group (262.22 ± 7.96%). Based on this study, it was concluded that cinnamaldehyde and berberine loaded CA-BER-ME-Gel can be an effective prescription alternative for the treatment of acne vulgaris.

Distinct roles of programmed cell death ligands 1 and 2, based on the type of immunity

Abstract Programmed cell death-1 (PD-1) is an immuno-regulatory receptor binding to two ligands, PD-L1 and PD-L2, which are expressed on stimulated antigen presenting cells (APCs). Isolated APCs such as macrophages (Mϕ), dendritic cells (DC), and B cells were cultured with interferon (IFN)-γ, interleukin (IL)-4, and IL-17A, respectively, followed by PD-L1/L2 expression analysis using flow cytometry. Upon stimulation by IFN-γ on all APCs and IL-17A on Mϕ and B cells, respectively, strong and moderate upregulation of PD-L1 expression was observed over unstimulated controls. Only stimulation by IL-4 could upregulate PD-L2 expression on all APCs. Therefore, experiments were performed on murine models, causing 2,4-dinitrofluorobenzene (DNFB)-induced contact hypersensitivity (CHS), fluorescein isothiocyanate (FITC)-induced CHS, and imiquimod (IMQ)-induced psoriasis-like dermatitis, in order to trigger IFN-γ-mediated T helper (Th)1, IL-4-mediated Th2, and IL-17A-mediated Th17-type responses, respectively. In both Th1 and Th17-type immunity models, ear thickness changes in PD-L1 deficient (PD-L1−/−) mice were more severe than in wild-type (WT) and PD-L2 deficient (PD-L2−/−) mice (PD-L1−/−: 26.6±3.0, WT: 12.0±1.8 and PD-L2−/−: 12.3±1.9 [μm] in DNFB-induced CHS; 9.5±0.7, 6.2±0.4 and 5.7±0.6 [μm] in IMQ-induced psoriasis-like dermatitis [P &amp;lt; 0.01 by the U test], respectively). Thickness changes in PD-L2−/− were more severe than those in the WT and PD-L1−/− mice in the Th2-type immunity model (PD-L2−/−: 10.8±1.0, WT: 3.8±0.6 and PD-L1−/−: 3.6±0.6 [μm], [P &amp;lt; 0.01 by the U test], respectively). Collectively, PD-L1 has predominant roles in Th1 and Th17-type immunity; similarly, PD-L2 for Th2-type immunity.

The neuropeptide, neurokinin A, has divergent effects on mast cell function

Abstract Mast cells (MCs) play a critical role in initiation of cutaneous type 2 immune responses. In the cutaneous microenvironment, sensory nerve fibers form a cohesive unit with MCs. In this unit, MC function is likely affected by neuropeptides, such as neurokinin A (NKA). NKA is abundant in the skin, but the affects of NKA on cutaneous MCs have not been extensively reported. NKA signals preferentially through the neurokinin-2 receptor (NK2R). We hypothesized that NKA/NK2R interactions influence cutaneous MC function. The effect of NKA on MCs was evaluated in vivo using the MC/IgE-specific passive cutaneous anaphylaxis (PCA) model. NKA reduced IgE-initiated changes in ear thickness. Likewise, NK2R antagonism enhanced IgE-initiated ear thickness in the PCA model. Intracellular pathways mediating the effects of NKA were delineated using bone marrow derived MCs (BMMCs). BMMCs expressed NK2R protein constitutively and NK2R expression was up-regulated by IgE. Functionally, NKA increased the percentage of IgE-activated MCs releasing granules but repressed transcription and release of the type 2 cytokines, IL-4 and IL-13. Mechanistically, IgE-mediated granule release has been attributed to increased STAT3 activation, whilst IL-4 and IL-13 transcription in MCs is attributed to STAT5 activation. NKA augmented STAT3 phosphorylation in IgE-activated MCs but reduced STAT5 phosphorylation. In sum, NKA modifies IgE-initiated STAT3 and STAT5 activity, granule release and cytokine transcription in MCs in vitro and inhibits MC function in vivo. These data demonstrate a role for NKA in the regulation of MC processes involved in type 2 mediated allergic disorders.

Receptor Mincle promotes skin allergies and is capable of recognizing cholesterol sulfate

Sterile (noninfected) inflammation underlies the pathogenesis of many widespread diseases, such as allergies and autoimmune diseases. The evolutionarily conserved innate immune system is considered to play a key role in tissue injury recognition and the subsequent development of sterile inflammation; however, the underlying molecular mechanisms are not yet completely understood. Here, we show that cholesterol sulfate, a molecule present in relatively high concentrations in the epithelial layer of barrier tissues, is selectively recognized by Mincle (Clec4e), a C-type lectin receptor of the innate immune system that is strongly up-regulated in response to skin damage. Mincle activation by cholesterol sulfate causes the secretion of a range of proinflammatory mediators, and s.c. injection of cholesterol sulfate results in a Mincle-mediated induction of a severe local inflammatory response. In addition, our study reveals a role of Mincle as a driving component in the pathogenesis of allergic skin inflammation. In a well-established model of allergic contact dermatitis, the absence of Mincle leads to a significant suppression of the magnitude of the skin inflammatory response as assessed by changes in ear thickness, myeloid cell infiltration, and cytokine and chemokine secretion. Taken together, our results provide a deeper understanding of the fundamental mechanisms underlying sterile inflammation.

Detection of local inflammation induced by repeated exposure to contact allergens by use of IVIS SpectrumCT analyses.

Contact allergy is characterized by local skin inflammation that, in some cases, can result in systemic immune activation. To investigate whether IVIS SpectrumCT analyses can be used to detect the immune response induced by contact allergens. Mice were repeatedly exposed to vehicle or allergens on the ears. The local and systemic responses were analysed at different times with the ProSense 750 FAST probe in IVIS SpectrumCT measurements. In addition, changes in ear thickness, cytokine profile in the skin and immunological phenotype in the draining lymph nodes and spleen were determined. Local inflammation was detected by ProSense 750 FAST and correlated with changes in ear thickness, cytokine profile and immunological phenotype following exposure to the strong contact allergen 2,4-dinitrofluorobenzene. Analysis of the systemic response with ProSense 750 FAST did not show any difference between allergen-exposed and control mice, although fluorescence-activated cell sorting analysis of the spleen showed increased numbers of γδ T cells and CD11b+ CD11c+ MHCII+ cells in allergen-treated mice. IVIS SpectrumCT analyses with ProSense 750 FAST as the probe can be used to detect local immune responses induced by contact allergens.

Pulichalconoid B suppresses experimental dermatitis in mice

Atopic dermatitis is a chronic and recurring inflammatory disease of the skin. This study aimed to evaluate the anti-inflammatory properties of pulichalconoid B, a chalcone that we have isolated from the plant Pulicaria incisa. The in vivo activity of pulichalconoid B was evaluated using the oxazolone model of cutaneous dermatitis and its in vitro activity was studied using isolated splenocytes. Changes in ear thickness, myeloperoxidase activity and production of different cytokines were considered to be indicators of dermal inflammation. Mice that were treated with pulichalconoid B showed a significantly decreased response to delayed-type hypersensitivity induced by oxazolone. Pulichalconoid B significantly reduced oxazolone-induced ear edema, inhibited myeloperoxidase activity and downregulated several cytokines. Pulichalconoid B also inhibited the secretion of cytokines from activated splenocytes. The data suggest that pulichalconoid B may be useful in treating skin inflammations.

Topical application of a platelet activating factor receptor agonist suppresses phorbol ester-induced acute and chronic inflammation and has cancer chemopreventive activity in mouse skin.

Platelet activating factor (PAF) has long been associated with acute edema and inflammatory responses. PAF acts by binding to a specific G-protein coupled receptor (PAF-R, Ptafr). However, the role of chronic PAF-R activation on sustained inflammatory responses has been largely ignored. We recently demonstrated that mice lacking the PAF-R (Ptafr-/- mice) exhibit increased cutaneous tumorigenesis in response to a two-stage chemical carcinogenesis protocol. Ptafr-/- mice also exhibited increased chronic inflammation in response to phorbol ester application. In this present study, we demonstrate that topical application of the non-hydrolysable PAF mimetic (carbamoyl-PAF (CPAF)), exerts a potent, dose-dependent, and short-lived edema response in WT mice, but not Ptafr -/- mice or mice deficient in c-Kit (c-Kit W-sh/W-sh mice). Using an ear inflammation model, co-administration of topical CPAF treatment resulted in a paradoxical decrease in both acute ear thickness changes associated with a single PMA application, as well as the sustained inflammation associated with chronic repetitive PMA applications. Moreover, mice treated topically with CPAF also exhibited a significant reduction in chemical carcinogenesis. The ability of CPAF to suppress acute and chronic inflammatory changes in response to PMA application(s) was PAF-R dependent, as CPAF had no effect on basal or PMA-induced inflammation in Ptafr-/- mice. Moreover, c-Kit appears to be necessary for the anti-inflammatory effects of CPAF, as CPAF had no observable effect in c-Kit W-sh/W-sh mice. These data provide additional evidence that PAF-R activation exerts complex immunomodulatory effects in a model of chronic inflammation that is relevant to neoplastic development.

Supplementing Milk Replacer with Omega-3 Fatty Acids from Fish Oil on Immunocompetence and Health of Jersey Calves

Fifty-one Jersey bull calves (5±1 d old) were assigned to 1 of 3 milk replacers to determine the effects of increasing doses of n-3 fatty acids from fish oil on immunocompetence and health. All calves were fed a 22.5% crude protein and 18% lipid industry standard milk replacer supplemented with an additional 2% fatty acids. The 3 treatments differed only in the supplemental lipid source and included a 3:1 mix of corn and canola oils; a 1:1 blend of fish oil and the 3:1 mix of corn and canola oils; and fish oil only. All treatments were supplemented with 150mg of vitamin E/kg of milk replacer. Body weight, height at withers, and length between withers and pins were measured weekly. Fecal and respiratory scores were recorded multiple times daily, and peripheral blood samples were collected on d 0, 7, 14, 21, and 42 for hematologic and metabolic analyses. Immunocompetence of calves was evaluated in vitro by the ability of neutrophils and monocytes to phagocytose Escherichia coli and produce an oxidative burst and in vivo as the change in ear thickness after an intradermal injection of phytohemagglutinin-P, and the primary and secondary humoral responses to ovalbumin. Production and health parameters were unaffected by treatments. There were no significant treatment or treatment × time effects on phagocytosis; however, there was a significant quadratic response for the percentage of neutrophils producing an oxidative burst. Fish oil did not affect the change in ear thickness in response to phytohemagglutinin-P. There was also no treatment effect on the primary IgG humoral response to ovalbumin, but there was a significant quadratic treatment effect on the secondary IgG response. Adding fish oil to milk replacer altered various immune responses, and the effect was dose-dependent; however, neither production performance nor indices of health were altered when fish oil replaced 5 to 10% of the fatty acids in milk replacer.

The helicase primase inhibitor, BAY 57-1293 shows potent therapeutic antiviral activity superior to famciclovir in BALB/c mice infected with herpes simplex virus type 1

BAY 57-1293 represents a new class of potent inhibitors of herpes simplex virus (HSV) that target the virus helicase primase complex. The present study was conducted using the zosteriform infection model in BALB/c mice. The helicase primase inhibitor, BAY 57-1293 was shown to be highly efficacious in this model. The beneficial effects of therapy were obtained rapidly (within 2 days) although the onset of treatment was delayed for 1 day after virus inoculation. The compound given orally, or intraperitoneally once per day at a dose of 15 mg/kg for 4 successive days was equally effective or superior to a much higher dose of famciclovir (1 mg/ml, i.e. approximately 140–200 mg/kg/day) given in the drinking water for 7 consecutive days, which, in our hands, is the most effective method for administering famciclovir to mice. In contrast to the vehicle-treated infected mice, all mice that received antiviral therapy looked normal and active with no mortality, no detectable loss of weight and no marked change in ear thickness. BAY 57-1293 and famciclovir reduced the virus titers in the skin to below the level of detection by days 3 and 7 post infection, respectively. In both BAY 57-1293 and famciclovir-treated mice, infectious virus titers in the ear pinna and brainstem remained below the level of detection. Consistent with these findings, BAY 57-1293 also showed a potent antiviral effect in an experiment involving a small number of severely immunocompromised athymic-nude BALB/c mice.

Cutaneous anaphylaxis in mice to Der p 2 independent of IgE and IgG 1

Rationale We have recently shown that immunoglobulin free light chains (IgLC) can elicit immediate hypersensitivity-like responses (Nature Medicine 8, 694, 2002). In this study, we investigated if cutaneous anaphylaxis to a major house dust mite (HDM) antigen could be induced by IgLC. Methods Mice were intradermally sensitized with two Der p 2-specific IgLC to different epitopes. At twenty hours mice received an intravenous challenge with HDM extract or recDer p 2 and changes in ear thickness were monitored. To investigate if anaphylaxis could be inhibited by a specific IgLC antagonist, F991, mice were injected i.p. with 50 microgram F991 at the time of sensitization. Results Intracutaneous injection of 2C5 and A4 followed by intravenous challenge with recDer p or HDM extract resulted in a rapid development of ear swelling reaching a maximum at 10 to 30 min after challenge. At 60 min post challenge, the ear thickness returned to basal levels. The PCA induced by crosslinking of Der p 2-specific Ig light chains could be completely prevented by F991. Conclusions The role of IgE in atopic dermatitis (AD) induced by aeroallergens remains controversial. Total or Ag-specific IgE does not always correlate with clinical symptoms. In several studies a discordance between the results from the atopy patch test or skin prick test and the levels of IgE specific for HDM were reported. We show that Der p 2-specific IgLC can elicit an IgE/IgG 1-independent cutaneous anaphylaxis. Our study suggests that induction of immediate hypersensitivity-like responses via IgLC might be responsible for positive skin reactions in non-atopic or intrinsic AD.

The expression of murine eutaneous late phase reaction requires both IgE antibodies and CD4 T eells

Exposure of atopic patients to a specific allergen evokes an immediate response which is followed, in many cases, by a late phase reaction (LPR) some hours later. Here we have examined the immunological mechanisms required for the expression of cutaneous LPR in mice. BALB/c mice were immunized by i.p. injection of ovalbumin (OVA) and alum actively or by i.v. injection of anti-OVA IgE monoclonal antibody (mAb) passively. After challenge by intradermal injection of OVA into ears, the changes in ear thickness, the number of eosinophils, and the levels of IL-4 and IFN-gamma protein at the site of antigen challenge were examined. Actively immunized mice developed a biphasic response at the site of OVA injection, while mice passively immunized with IgE anti-OVA mAb displayed a strong early response but no LPR. Cell transfer experiments using BALB/c nu/nu mice revealed that both OVA-specific IgE mAb and OVA-primed CD4 T cells were required to evoke LPR. Moreover, LPR was associated with increased levels of IL-4 production concomitant with reduced IFN-gamma production and was abolished by pretreatment with anti-IL-4 neutralizing mAb. It is suggested that murine cutaneous LPR against OVA is a type 2 inflammatory response in which both IgE antibodies and CD4 T cells play an obligatory role.