IgE-mediated Cutaneous Reaction Research Articles

Effects of a selective casein kinase 1δ and ε inhibitor on FcεRI expression and IgE-mediated immediate-type cutaneous reactions in dogs.

The molecular clock network in mast cells has been shown to be a factor responsible for circadian regulation of allergic inflammation. PF670462 is a selective inhibitor of casein kinase 1δ and ε (CK1δ/ε) that control the posttranslational modification of clock proteins. The aims of this study were to evaluate the effects of PF670462 on gene and protein expression of FcεRI, the high-affinity IgE receptor, in canine mast cells and on IgE-mediated immediate-type cutaneous reactions in dogs. PF670462 decreased mRNA expression of FcεRIα and β, but not γ, and protein expression of FcεRI in a canine mast cell line. Furthermore, PF670462 suppressed IgE-mediated immediate-type cutaneous erythema in dogs. These findings indicate that CK1δ/ε function as regulators for FcεRI expression and IgE-mediated cutaneous reactions in dogs.

Protein tyrosine phosphatase 1B (PTP1B) is dispensable for IgE-mediated cutaneous reaction in vivo

Mast cells play a critical role in allergic reactions. The cross-linking of FcεRI-bound IgE with multivalent antigen initiates a cascade of signaling events leading to mast cell activation. It has been well-recognized that cross linking of FcεRI mediates tyrosine phosphorylation. However, the mechanism involved in tyrosine dephosphorylation in mast cells is less clear. Here we demonstrated that protein tyrosine phosphatase 1B (PTP1B)-deficient mast cells showed increased IgE-mediated phosphorylation of the signal transducer and activator of transcription 5 (STAT5) and enhanced production of CCL9 (MIP-1γ) and IL-6 in IgE-mediated mast cells activation in vitro. However, IgE-mediated calcium mobilization, β-hexaosaminidase release (degranulation), and phosphorylation of IκB and MAP kinases were not affected by PTP1B deficiency. Furthermore, PTP1B deficient mice showed normal IgE-dependent passive cutaneous anaphylaxis and late phase cutaneous reactions in vivo. Thus, PTP1B specifically regulates IgE-mediated STAT5 pathway, but is redundant in influencing mast cell function in vivo.

Involvement of cannabinoid CB 2 receptors in the IgE-mediated triphasic cutaneous reaction in mice

Involvement of cannabinoid CB 2 receptors in the IgE-mediated cutaneous reaction was investigated. Epicutaneous challenge with 2,4-dinitrofluorobenzene caused a triphasic swelling in the ear of BALB/c and C57BL/6 mice passively sensitized with anti-dinitrophenol IgE. Peak responses of the ear swelling appeared at 1 h, 24 h, and 8 days after the challenge in both strains of mice. In contrast, cannabinoid CB 2 receptor-deficient mice failed to exhibit the obvious triphasic ear swelling observed in wild-type mice. Oral administration of cannabinoid CB 2 receptor antagonist/inverse agonists [ N-(benzo[1,3]dioxol-5-ylmethyl)-7-methoxy-2-oxo-8-pentyloxy-1,2-dihydroquinoline-3-carboxamide] (JTE-907) and { N-[(1S)- endo-1,3,3-trimethylbicyclo[2,2,1]heptan-2yl]5-(4-chloro-3-methyl-phenyl)-1-(4-methylbenzyl)pyrazole-3-carboxamide} (SR144528) at doses of 0.1–10 mg/kg significantly and dose-dependently suppressed all three phases of ear swelling in BALB/c mice. Interestingly, epicutaneous treatment with an ether-linked analogue of endogenous cannabinoids, 2-arachidonoylglycerol, caused an ear swelling that could be detected at 1 h, 24 h, and 8 days after treatment of both BALB/c and C57BL/6 mice. These results suggest that cannabinoid CB 2 receptors are involved in induction of the triphasic cutaneous reaction mediated by IgE, and that cannabinoid CB 2 receptor antagonist/inverse agonists may serve as anti-allergic agents in the treatment of allergic dermatitis.

Differential Effect of Antiallergic Drugs on IgE-Mediated Cutaneous Reaction in Passively Sensitized Mice

We investigated the effect of several antiallergic agents on murine IgE-mediated biphasic cutaneous reaction. Mice were passively sensitized by an intravenous injection of monoclonal anti-dinitrophenol (anti-DNP) IgE antibody. Skin reaction was elicited by an epicutaneous challenge of dinitrofluorobenzene (DNFB) and occurred biphasically with immediate-phase response (IPR) and late-phase response (LPR) at 1 and 24 h, respectively. Classical histamine H1 receptor antagonists and some chemical mediator-release inhibitors, such as diphenhydramine and terfenadine, inhibited IPR, but not LPR. In contrast, leukotriene B₄ (LTB₄) receptor antagonist (ONO-4057) inhibited LPR only. Antagonists for LTC₄, D₄, E₄ receptor (ONO-1078) and platelet-activating factor (PAF) receptor (Y-24180) significantly inhibited both IPR and LPR, similarly to prednisolone. We recently found that a third-phase inflammatory reaction with marked infiltration of eosinophils (named very-late-phase response; vLPR), which is supposed to be a more important reaction in allergic diseases, was induced, peaking at day 8 following IPR and LPR in this model. The effect of these drugs on the triphasic skin reaction can be scored based on efficacy against IPR / LPR / vLPR; +/+/+ (prednisolone, a PAF receptor antagonist Y-24180, cyclosporin A and FK-506), +/–/– (diphenhydramine), +/+/– (azelastine and LT receptor antagonist ONO-1078), and –/+/+ (an LTB₄ receptor antagonist ONO-4057). Thus the inhibitory effect on vLPR as well as LPR may relate to the inhibition of eosinophil function mediated by LTB₄ and PAF and/or T cells. These findings may provide the basis for a treatment modality using various antiallergic agents in allergic disease.

A third-phase cutaneous (very late phase) response after elicitation with dinitrofluorobenzene in passively or actively sensitized mice

Previous studies have reported that the mice passively sensitized with anti-dinitrophenol (DNP) IgE antibody exhibited IgE-mediated cutaneous reaction with an immediate phase response (IPR) at 1 h and a late phase response (LPR) at 24 h after the challenge of dinitrofluorobenzene (DNFB). We found that the third-phase inflammatory reaction with intense and persisting infiltration of eosinophils, named ‘very late phase reaction (vLPR)’, was induced following IPR and LPR in response to DNFB in actively and passively sensitized mice, and that the peak response of vLPR was at 8 days after the challenge. This reaction was slightly observed in non-sensitized mice. Since the accumulation of eosinophils in vLPR was markedly observed when compared with that of LPR at 24 h, the vLPR may be an important reaction in allergic diseases. The development of vLPR was partly decreased in mast cell-deficient WBB6F1-W/Wv mice and was absent in T cell-deficient BALB/c-nu/nu mice in passive sensitization. These results indicate that the vLPR in the triphasic cutaneous reaction may be mainly mediated by T cells and partially by mast cells and/or IgE antibody, and consequently lead to an intense ear swelling accompanying massive infiltration of eosinophils.

Inhibitory mechanisms of glycoprotein fraction derived from Miscanthus sinensis for the immediate phase response of an IgE-mediated cutaneous reaction.

We investigated the inhibitory effect of the glycoprotein fraction (fraction 2) extracted from Miscanthus sinensi ANDERSSON (M. sinensis) on biphasic cutaneous reactions in mice passively sensitized with IgE. Biphasic skin reactions with peak responses at 1 (IPR, immediate phase reaction) and 24 h (LPR, late phase reaction) were caused by passive sensitization with an anti-dinitrophenol IgE monoclonal antibody (anti-DNP IgE mAb) followed by an epicutaneous challenge of 0.1% dinitrofluorobenzene (DNFB) in 100% ethanol. Intraperitoneal injection of fraction 2 before the DNFB challenge significantly inhibited the biphasic ear swelling response in passively sensitized mice in a dose-dependent manner (1-30 mg/kg). We also found that fraction 2 was effective at inhibiting the vascular permeability in mouse ear induced by an injection of compound 48/80, histamine or serotonin. In addition, fraction 2 inhibited scratching behavior as well as ear edema observed within 2 h after DNFB challenge. Marked inhibition was observed in both passively sensitized and non-sensitized mice. The locomotor activity of mice was also reduced by the administration of fraction 2 as well as by diphenhydramine. These results suggest that the inhibitory effect of glycoprotein fraction 2 of M. sinensis on an IgE-mediated allergic inflammatory reaction is due to the protection of mediator-induced vascular permeability and that in addition to the inhibition of an inflammatory reaction, a sedative action is responsible for the inhibition of allergy-induced scratching responses.

Effect of Y-24180, an antagonist at platelet-activating factor (PAF) receptors, on IgE-mediated cutaneous reactions in mice

We examined the effect of Y-24180, a potent platelet-activating factor (PAF) antagonist, on IgE-mediated cutaneous reactions in mice. Female BALB/c mice were used. Drugs were orally administered 1 h before a dinitrofluorobenzene (DNFB) challenge or 1 h before and 12 h after the challenge. Biphasic increase in ear thickness, with peak responses at 1 h (immediate phase reaction, IPR) and 24 h (late phase reaction, LPR) after the DNFB challenge, were induced in mice which had been passively sensitized with monoclonal anti-dinitrophenyl IgE antibody 24 h before the DNFB challenge. Ear thickness was measured with a dial thickness gauge. Y-24180, WEB2086, ketotifen, and suplatast suppressed the IPR. Y-24180 also suppressed the LPR when administered once at 10 mg/kg or twice at 1 to 10 mg/kg. WEB2086 suppressed the LPR only when administered twice. However, ketotifen and suplatast did not suppress the LPR even when administered twice. Single administration of prednisolone significantly suppressed both the IPR and LPR. These results indicate that PAF may be involved in the induction of biphasic cutaneous reactions mediated by IgE, and Y-24180 is more effective compared with WEB2086 in this model. It is possible that the difference in the effectiveness between Y-24180 and WEB2086 depends on the persistence of those activities.

Topical glucocorticoid augments IgE-mediated passive cutaneous anaphylaxis in Balb/C mice and mast cell deficient WBB6F1 v/v mice.

In a last decade, new types of skin manifestations have been recognized in atopic dermatitis especially in Japan. They are frequently observed in adult patients with atopic dermatitis after a long-standing steroid ointment and termed adult type-atopic dermatitis. To clarify whether topical glucocorticoid (GC) modulates cutaneous inflammatory reactions in addition to known anti-inflammatory effect, we have examined the effect of long-term application of topical GC on IgE-mediated murine cutaneous reactions. Fifty microlitres of diflucortolone valerate (1 mg/mL), prednisolone valerateacetate (3 mg/mL), or triamcinolone acetonide (1 mg/mL) were applied seven times on alternate day, to the flank skin of mice. On day 12 when mice received the seventh application of GC, each mouse was given an intravenous application of IgE anti-DNP antibody (PCA titre > x 2560) 1 h before the skin test with 0.15% DNFB in acetone:olive oil (4:1) on the right pinna. The left pinna was painted with a vehicle as a control. Increased ear thickness was measured at 1, 4, 24, 48 and 72 h to assess the augmentry effect of GC. Topical application of GC (50 microg diflucortolone valerate in ethanol) on the flank skin seven times on alternate days, augmented expression of passive cutaneous anaphylaxis reaction on the ear skin induced by intravenous applications of monoclonal IgE anti-DNP antibody and following the challenge test. In contrast, topical application of GC inhibited the reactions when applied on the challenged sites. Several types of GC, but not vitamin D3, augmented the skin reactions and these augmented reactions persisted for 72 h when control skin reactions subsided. GC induced a late phase but not an early phase cutaneous reaction in mast cell deficient WBB6F1 v/v mice by IgE anti-DNP antibody. Long-term application of topical GC might modulate local cutaneous immune response and augment IgE-mediated cutaneous reactions. Fc epsilon R(+) cells other than mast cell might be involved in the IgE-mediated late-phase reaction.

TNF-alpha participates in an IgE-mediated cutaneous reaction in mast cell deficient, WBB6F1-W/Wv mice.

The participation of tumor necrosis factor-alpha (TNF-alpha) in a IgE-mediated cutaneous reaction in WBB6F1-W/Wv (W/Wv), mast cell deficient, mice and the effect of prednisolone on this cutaneous reaction were investigated. Mice were passively sensitized by an intravenous injection of monoclonal anti-dinitrophenol (DNP) IgE, and their ears challenged epicutaneously with dinitrofluorobenzene 24 h later. The cutaneous reaction estimated by ear thickness reached a peak 48-72 h after the antigen challenge. A monoclonal anti-tumor necrosis factor (TNF)-alpha antibody inhibited the IgE-mediated cutaneous reaction. An increase of TNF-alpha mRNA was demonstrated 4 h after the application of antigen by the reverse transcriptase-polymerase chain reaction. The injection of recombinant murine TNF-alpha induced a cutaneous reaction which peaked at 24 h in nonsensitized mice. Prednisolone at doses of 3 to 10 mg/kg clearly inhibited the IgE-mediated cutaneous reaction, however, it did not affect the expression of TNF-alpha-mRNA. Prednisolone at doses of 1 to 10 mg/kg clearly inhibited the TNF-alpha-induced cutaneous reaction. These results suggest that TNF-alpha plays a role in the IgE-mediated cutaneous reaction in W/Wv mice and that prednisolone inhibits the cutaneous reaction at least in part by inhibiting the action of TNF-alpha.

Effect of mast cell modulators on IgE-mediated murine biphasic cutaneous reactions.

Hapten-specific and mast cell-dependent biphasic cutaneous reactions were induced by intravenous application of anti DNP-IgE antibodies and a subsequent skin test. These reactions were also demonstrated in SCID mice, which indicates that T cell-mediated immunity might not be involved in these IgE-mediated cutaneous reactions. Simultaneous application of anti histaminics did not suppress these reactions significantly, while several immunomodulators, such as azelastine, FK506, and prednisolone, significantly inhibited both early and late phase reactions except for the failure of FK506 to inhibit the early reaction. Anti-VCAM-1 antibody and anti-tumor necrosis factor-alpha (TNF alpha) antibody but not anti-IL 5 antibody showed similar suppressive effects on both early and late phase reactions. Mast cell and inflammatory cells other than T cells are thought to play an important role in these IgE-induced biphasic reactions. TNF alpha and/or VCAM-1 are required for tissue accumulation of inflammatory cells in this system.

Topical vitamin D3 downregulates IgE-mediated murine biphasic cutaneous reactions.

Hapten-specific and mast-cell-dependent biphasic (immediate and delayed-onset) cutaneous reactions were induced in a murine model by intravenous injection of anti-DNP IgE antibodies followed by a skin test. Four daily applications of topical 1 alpha,24(OH)2D3 ointment significantly inhibited both the immediate and the delayed-onset cutaneous reactions in a dose-dependent fashion, as well as the croton-oil-induced cutaneous reaction and DNFB contact sensitivity reaction. 1 alpha,24(OH)2D3 itself did not show any sensitizing or irritant potential. The inhibitory effect of 1 alpha,24(OH)2D3 on these reactions was limited to the application site and no systemic effect was observed. Another vitamin D3 analog 1 alpha,25(OH)2D3, also showed an inhibitory effect on IgE-mediated cutaneous reactions. These results suggest that topically applied 1 alpha,24(OH)2D3 might modulate IgE-mediated cutaneous reactions and could thus be useful in the treatment of certain human cutaneous disorders other than psoriasis and related keratinizing disorders.

Rhesus Monkey Airway Responses to Substance P

The tachykinin, substance P (SP), was used as a model to study airway and cutaneous responses in a group of normal rhesus monkeys or animals with IgE-mediated cutaneous reactions to Ascaris antigen (AA) alone or animals with both IgE-mediated cutaneous and airway responses to AA. Aerosolized SP (10 mg/ml) resulted in airway responses qualitatively similar in duration and pulmonary function abnormalities to AA. These SP airway responses were most closely associated with the presence of IgE antibody to AA and were not increased in animals with airway reactivity to AA. A dose response to aerosolized SP can be established. An enkephalinase inhibitor, thiorphan, did not potentiate SP airway responses but appeared to potentiate SP cutaneous responses in some animals. SP cutaneous reactions could be demonstrated and did not correlate with the IgE-mediated cutaneous dilution titers to AA.