anti-DNP IgE Research Articles

Role of TGF-β1 on the IgE-Dependent Anaphylaxis Reaction

Abstract TGF-β1 is a member of a family of polypeptide factors that control proliferation, differentiation, chemotaxis, and other functions in many cell types. TGF-β1 has been shown to inhibit many immunologic functions. However, here we report that TGF-β1 has an important role in the elicitation of IgE-dependent allergic reactions. The synthetic antisense TGF-β1 oligonucleotides dose-dependently inhibit passive cutaneous anaphylaxis (PCA) reaction and histamine release from the mast cells activated by anti-DNP IgE in rats. The level of cAMP in mast cells, when antisense TGF-β1 oligonucleotides was added, significantly increased ∼7-fold compared with that of basal cells. The antisense TGF-β1 oligonucleotides also had a significant inhibitory effect on anti-DNP IgE-induced TNF-α release from mast cells. In situ hybridization analysis showed that the PCA reaction sites treated with antisense TGF-β1 oligonucleotides exhibited no detectable levels of TGF-β1 and l-histidine decarboxylase mRNA after anti-DNP IgE stimulation, whereas the PCA reaction sites treated with sense TGF-β1 oligonucleotides possessed significant amounts of their mRNA. Additionally, neutralizing Ab to TGF-β1 blocked the PCA reaction significantly, but its Ab did not inhibit peritoneal mast cell-released histamine upon treatment with anti-DNP IgE. Our results suggest that TGF-β1 is critical to the development of IgE-dependent anaphylaxis reactions.

Involvement of Farnesyl Protein Transferase (FPTase) in FcεRI-Induced Activation of RBL-2H3 Mast Cells

Changes in farnesyl protein transferase (FPTase) activity and FPTase β-subunit protein levels were determined in IgE-sensitized RBL-2H3 mast cells in response to polyvalent antigen administration. Ten minutes after the addition of DNP modified BSA to mast cells, whose high affinity receptor for IgE (FcεRI) contained bound anti-DNP IgE, FPTase specific activity increased by 54 ± 28%. Time course studies showed FPTase specific activity doubled during a 20- to 30-min period after antigen-induced cell aggregation. Also, an increase in FPTase β-subunit protein during this time (∼30%) was observed; this protein increase was not accompanied by a similar increase in FPTase β-subunit m-RNA levels. The FcεRI aggregation had no significant effect on the activities of other enzymes involved with farnesyl diphosphate (FPP) metabolism: FPP synthase, isopentenyl diphosphate isomerase, geranylgeranyl protein transferase, and squalene synthase. Specific inhibition of FPTase activity by manumycin was studied to determine what role FPTase plays in mast cell activation. Manumycin profoundly inhibited hexosaminidase release in activated cells, indicating FPTase is required for signal transduction involved with protein exocytosis from RBL-2H3 mast cells.

Lavender oil inhibits immediate-type allergic reaction in mice and rats.

We studied the effects of lavender oil on mast cell-mediated immediate-type allergic reactions in mice and rats. Lavender oil (1:500, 1:100, 1:10, 1:1, 1:0) inhibited concentration-dependently mast cell-dependent ear swelling response induced by compound 48/80 in mice by both topical and intradermal application. Lavender oil (1:500, 1:100, 1:10, 1:1, 1:0) inhibited concentration-dependently passive cutaneous anaphylaxis induced by anti-dinitrophenyl (DNP) IgE in rats by both topical and intradermal application. Lavender oil (1:500, 1:100, 1:10, 1:1, 1:0) also inhibited concentration-dependently the histamine release from the peritoneal mast cells by compound 48/80 or anti-DNP IgE. Moreover, lavender oil (1:1000, 1:100, 1:10, 1:0) had a significant inhibitory effect on anti-DNP IgE-induced tumour necrosis factor-alpha secretion from peritoneal mast cells. These results indicate that lavender oil inhibits immediate-type allergic reactions by inhibition of mast cell degranulation in-vivo and in-vitro.

MAGNOLIAE FLOS INHIBITS MAST CELL-DEPENDENT IMMEDIATE-TYPE ALLERGIC REACTIONS

The mast cell plays a pivotal role in initiating allergic response by secreting intracytoplasmic granular mediators such as histamine. Magnoliae flos has been used for the treatment of allergic disease in Korea. However, its effect in experimental models remains unknown. The present report describes an inhibitory effect of Magnoliae flos on mast cell-mediated immediate-type allergic reactions. Topical application of compound 48/80 can induce an ear swelling response in normal (WBB6F1-+/+) mice but not in the congenic mast cell-deficient WBB6F1-W/Wvmice. Magnoliae flos inhibited concentration-dependently mast cell-dependent ear swelling response induced by compound 48/80 by topical application. Magnoliae flos inhibited concentration-dependently passive cutaneous anaphylaxis induced by anti-dinitrophenyl (DNP) IgE in rats by topical application. Magnoliae flos also inhibited concentration-dependently the histamine release from the rat peritoneal mast cells by compound 48/80 or anti-DNP IgE. Moreover, Magnoliae flos had a significant inhibitory effect on compound 48/80-induced systemic anaphylactic reaction. These results indicate that Magnoliae flos inhibits immediate-type allergic reactions by inhibition of mast cell degranulationin vivoandin vitro.

Eft of Schizonepeta Tenuifolia Extract On Mast Cell-Mediated Immediate-Type Hypersensitivity In Rats

We investigated the effect of an aqueous extract of Schizonepeta tenuifolia (STAE) on mast cell-mediated immediate-type hypersensitivity. STAE inhibited systemic allergic reaction induced by compound 48/80 in rats dose-dependently. STAE also inhibited plasma histamine levels induced by compound 48/80. STAE inhibited local allergic reaction activated by anti-dinitrophenyl (DNP) IgE. In addition, STAE does-dependently inhibited histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. However, STAE had a significant enhancing effect on anti-DNP IgE-induced tumor necrosis factor-α (TNF-α) production from RPMC. These results indicate that STAE inhibits immediate-type hypersensitivity and suggest that STAE can selectively activate the TNF-α production from RPMC.

The evaluation of antianaphylactic effect of Oryza sativa L. in rats.

This study was carried out to examine the effect of methanol extract of Oryza sativa L. (Dong-Jin in Korean, abbreviate as Os-DJ hereafter) on anaphylaxis. Os-DJ (10(-5) to 1 g/kg) dose-dependently inhibited systemic anaphylaxis induced by compound 48/80 in rats. When Os-DJ was pretreated at concentration ranging from 10(-5) to 1 g/kg, the serum histamine levels were reduced in a dose-dependent manner. Os-DJ (1 g/kg) also significantly inhibited local anaphylaxis activated by anti-dinitrophenyl (DNP) IgE. Moreover, Os-DJ dose-dependently inhibited the histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. These results indicate that Os-DJ possess antianaphylactic activity by inhibition of histamine release from mast cells in vivo and in vitro.

Eosinophil and IgE responses of IL-5 transgenic mice experimentally infected with Nippostrongylus brasiliensis.

Eosinophil and IgE responses of interleukin (IL)-5 transgenic and normal C3H/HeN mice were studied after experimental infection with Nippostrongylus brasiliensis (Nb). Intestinal worms were recovered at day 5 post-infection (PI), and numbers of total white blood cells (WBC) and eosinophils, and total serum IgE and anti-hapten (dinitrophenyl) (DNP) specific IgE titers, were measured at days 0, 14 and 21 PI. IL-5 mice appeared resistant to Nb infection showing a significantly lower worm recovery rate than normal mice (P < 0.05). Total WBC and eosinophil counts (/mm3) were significantly increased in Nb infected normal mice (P < 0.05), but unchanged (total WBC) or decreased (eosinophils) in IL-5 mice at day 21 PI. The total serum IgE level remarkably increased in normal mice, but only a little in IL-5 mice at days 14 and 21 PI. Priming with DNP brought about more remarkable increases of the total and anti-DNP specific IgE in normal mice than in IL-5 mice. The results show that IL-5 mice are resistant to Nb infection, and that eosinophil and IgE responses in these mice are not augmented by Nb infection.

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.

Inhibition of Mast Cell-Dependent Anaphylactic Reactions by the Pigment of Polygonum tinctorium (Chung-Dae) in Rats

1. The effect of the pigment obtained from the stem and leaf of Polygonum tinctorium Loar (PtP) on anaphylactic reactions was studied in rats. 2. PtP totally inhibited compound 48/80-induced anaphylactic shock with doses of 10 2 and 10 3 mg/kg. When PtP was pretreated at concentrations ranging from 10 −2 to 10 3 mg/kg, the serum histamine levels induced by compound 48/80 were reduced in a dose-dependent manner. 3. We also investigated the effect of PtP on mast cell-dependent passive cutaneous anaphylaxis (PCA) activated by anti-dinitrophenyl (DNP) IgE antibody. PtP potently inhibited PCA when administered orally, topically, intraperitoneally and intradermally. However, it did not show inhibitory activity when administered intravenously. 4. PtP inhibited dose dependently histamine release from rat peritoneal mast cells (RPMCs) induced by compound 48/80 and anti-DNP IgE. Moreover, the level of cAMP in RPMC, when PtP was added, significantly increased about 12-fold at 4 min compared with that of basal cells. 5. These results indicate that PtP may possess strong antianaphylactic activity and suggest that differences in bioavailability may cause differential activity following different administration routes.

Short Communication Inhibition of immediate type allergic reactions by the aqueous extract of Kum-Hwang-San

We studied the effect of aqueous extract of Kum-Hwang-San (KHS) on mast cell-mediated immediate type allergic reactions. KHS (1–100 μg\\site) inhibited concentration-dependently mast cell-dependent ear swelling response induced by compound 48\\80 (200 μg\\site) in mice by both topical and intradermal application. KHS (0.1–100 μg\\site) inhibited concentration-dependently passive cutaneous anaphylaxis induced by anti-dinitrophenyl (DNP) IgE in rats by both topical and intradermal application. KHS also inhibited concentration-dependently the histamine release from the rat peritoneal mast cells (RPMC) by compound 48\\80 and anti-DNP IgE. Moreover, KHS had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor- fn2 fn2 tumor necrosis factor- α; HSA: human serum albumin; ELISA: enzyme linked immunosorbent assay. (TNF-α) secretion from RPMC. These results indicate that KHS inhibits immediate type allergic reactions by inhibition of histamine release and TNF-α secretion from mast cells in vivo and in vitro.

Effect of Rehmannia glutinosa on immediate type allergic reaction

We investigated the effect of aqueous extract of Rehmannia glutinosa steamed root (RGAE) on the allergic reactions in vivo and in vitro. RGAE dose-dependently inhibited systemic allergic reaction induced by compound 48/80. When RGAE was pre-treated at the same concentrations with systemic allergic reaction test, the plasma histamine levels were reduced in a dose-dependent manner. RGAE dose-dependently inhibited skin allergic reaction activated by anti-dinitrophenyl (DNP) IgE. RGAE also dose-dependently inhibited the histamine release from the rat peritoneal mast cells (RPMC) by compound 48/80 or anti-DNP IgE. Moreover, RGAE had significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor- α production of RPMC. These results indicate that RGAE may be beneficial in the regulation of immediate type allergic reaction.

Inhibition of Anaphylaxis by Sulfasalazine in Rats

We studied the effect of sulfasalazine on anaphylaxis. Sulfasalazine dose-dependently inhibited systemic anaphylaxis induced by compound 48/80 in rats. Sulfasalazine also inhibited local anaphylaxis activated by anti-dinitrophenyl (anti-DNP) IgE. Moreover, sulfasalazine dose-dependently inhibited histamine release in the peritoneal mast cells activated by compound 48/80 or anti-DNP IgE. All of these effects were comparable to those of the disodium cromoglycate (reference drug) tested. When sulfasalazine was added, the level of cAMP in rat peritoneal mast cells transiently and significantly increased about 6-fold compared with that of basal cells. Our studies provide evidence that sulfasalazine may be beneficial in the treatment of anaphylaxis.

Inhibitory Effect of Mast Cell-Mediated Immediate-Type Allergic Reactions in Rats by Spirulina

We investigated the effect of spirulina on mast cell-mediated immediate-type allergic reactions. Spirulina dose-dependently inhibited the systemic allergic reaction induced by compound 48/80 in rats. Spirulina inhibited compound 48/80-induced allergic reaction 100% with doses of 100–1000 μg/g body weight, i.p. Spirulina (10–1000 μg/g body weight, i.p.) also significantly inhibited local allergic reaction activated by anti-dinitrophenyl (DNP) IgE. When rats were pretreated with spirulina at a concentration ranging from 0.01 to 1000 μg/g body weight, i.p., the serum histamine levels were reduced in a dose-dependent manner. Spirulina (0.001 to 10 μg/mL) dose-dependently inhibited histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. The level of cyclic AMP in RPMC, when spirulina (10 μg/mL) was added, transiently and significantly increased about 70-fold at 10 sec compared with that of control cells. Moreover, spirulina (10 μg/mL) had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-α production. These results indicate that spirulina inhibits mast cell-mediated immediate-type allergic reactions in vivo and in vitro.

Regulation of 10P2 murine mast cell proliferation and secretory function by stem cell factor or IL-9.

Mast cells are effectors of inflammatory responses. When triggered by immunological or nonimmunological mechanisms, mast cells release potent biological mediators from preformed stores and synthesize others de novo. In previous investigations from this laboratory, the signal transduction pathways of cloned 10P2 cytokine-independent mast cells were explored. Results suggested that 10P2 cells undergo activation-secretion coupling assessed as release of stored [14C]serotonin (5-HT) when challenged with IgE-specific antigen, influx of extracellular calcium, release of intracellular calcium stores, or by direct activation of protein kinase C isozymes. In the present investigations, cytokine proliferative effects and modulatory roles on release of stored [14C]5-HT have been explored. Following passive sensitization with anti-dinitrophenol (anti-DNP) IgE and challenge with DNP, mast cells released up to 32% of the stored [14C]5-HT. Pretreatment of cells with 10, 30, or 50 ng/ml stem cell factor (SCF) did not alter the response. SCF did not directly induce [14C]5-HT release. Pretreatment with 25 ng/ml interleukin-9 (IL-9) significantly potentiated the IgE-antigen release by 51.1%, 35.7%, or 31.6% when challenged with 3, 10 or 30 ng/ml DNP-HSA. Treatment of cells with 1-100 ng/ml SCF for 72 hr resulted in significantly enhanced proliferation whereas this did not occur when cells were treated with 1-100 ng/ml IL-9. Collectively, these results suggest that SCF alone has a proliferative effect, does not alter the IgE-specific antigen signal transduction pathway, and does not directly stimulate mast cell degranulation. In contrast, IL-9 potentiates the IgE-antigen signal transduction response but exerts no proliferative response. Reports of effects of orally administered cytokines are now beginning to emerge. This raises the possibility that cytokines may be a future therapeutic approach to treatment of allergic and nonallergic inflammatory diseases. The 10P2 cytokine-independent mast cell line may be a valuable adjunct to existing mast cell models as this avenue of drug discovery is explored.

Kinetics of Multivalent Antigen DNP-BSA Binding to IgE-FcεRI in Relationship to the Stimulated Tyrosine Phosphorylation of FcεRI

Abstract Multivalent DNP-BSA is commonly used to cross-link anti-DNP IgE bound to FcεRI to stimulate cellular responses, although key features of the binding process are unknown. Fluorescence quenching can be used to study the kinetics of DNP-BSA binding to FITC-IgE. We observe that DNP-BSA binds more slowly to IgE than does an equimolar amount of a monovalent DNP ligand, suggesting that the average effective number of DNP groups per BSA is less than one. The binding data are well described by a transient hapten exposure model in which most of the DNP groups are unavailable for binding but have some probability of becoming exposed and available for binding during the time of the binding measurement. Additional experiments indicate that, for suboptimal to optimal concentrations of DNP-BSA, most of the FITC fluorescence quenching on the cell surface is due to cross-linking events. With these concentrations at 15°C, the kinetics of FITC fluorescence quenching by DNP-BSA correlates with the kinetics of DNP-BSA-stimulated tyrosine phosphorylation of FcεRI. At 35°C, the phosphorylation kinetics are biphasic during the time period in which cross-linking continues to increase. Our results establish a quantitative relationship between the timecourse for cross-linking by multivalent Ag and FcεRI-mediated signaling, and they provide the means to predict the kinetics of cross-linking under a wide variety of conditions.

Topical glucocorticoid augments anti-DNP IGE mediated cutaneous reaction through increased IL1α production by keratinocytes

Topical glucocorticoid augments anti-DNP IGE mediated cutaneous reaction through increased IL1α production by keratinocytes

Effect of spikelets of Miscanthus sinensis on IgE-mediated biphasic cutaneous reaction in mice.

The effect of spikelets of Miscanthus sinensis Andersson (M. sinensis) on IgE-mediated biphasic cutaneous reactions was investigated in BALB/c mice. Mice were passively sensitized by an intravenous (i.v.) injection of monoclonal antidinitrophenol IgE antibody (anti-DNP IgE mAb), or actively by an intraperitoneal (i.p.) injection of DNP-derivatized ovalbumin (DNP-OVA) plus aluminium hydroxide gel (Alum) as an adjuvant. Skin reactions were elicited by an epicutaneous challenge of dinitrofluorobenzene (DNFB) and occurred biphasically with peak responses at 1 and 24 h in both animal models. The administrations of a nondialysable water extract of M. sinensis within 2 h before or after DNFB challenge via oral or i.p. route significantly inhibited the biphasic cutaneous reactions in passively and actively sensitized mice. The inhibitory effect was much stronger than those of a glucocorticoid, prednisolone, and histamine release inhibitor, amlexanox, as positive controls. The active component(s) was predominantly located in the glycoprotein fraction by gel chromatography. In the ears of DNFB-challenged mice, this fraction suppressed the accumulation of inflammatory cells, including mast cells and neutrophils/macrophages. In addition, the biphasic ear swelling was also improved by an administration of the glycoprotein fraction 24 h before active sensitization. These findings indicate that the glycoprotein fraction of M. sinensis was able to inhibit not only the IgE-mediated allergic inflammatory reaction but also the IgE formation. Thus, this fraction may be a useful antiallergic therapy.

Inhibitory effect of Anaphylactic Reaction of Sosiho-Tang

Mast cells synthesize and secrete chemical mediators which play an central role in anaphylactic reactions. Compound 48/80 is a condensation product of formaldehyde with paramethoxyphenylethylamine that reliably induces the release of chemical mediators in the mast cell granules. Aggregation of the high-affinity Fc receptor also stimulates the mast cells. The objective of the current study was to determine the effect of Sosiho-Tang (SS-Tang) on mast cell-mediated anaphylactic reaction. SS-Tang completely inhibited systemic anaphylaxis induced by compound 48/80 in mice. SS-Tang inhibited local anaphylaxis induced by anti-dinitrophenyl (DNP) IgE. In addition SS-Tang concentration-dependently inhibited histamine release in mast cells induced by compound 48/80 or anti-DNP IgE. These results indicate that SS-Tang may contain compounds with actions that inhibit mast cell degranulation.

Effect of Korean folk medicine ‘Chung-Dae-San' on mast cell-dependent anaphylactic reaction

We investigated the effect of the herbal formulation ‘Chung-Dae-San’ (CDS) on anaphylactic reactions. CDS inhibited compound 48/80-induced anaphylactic shock 100% with the dose of 10 0 g/kg body weight (BW). When CDS was given as pretreatment at concentrations ranging from 10 −4 to 10 0 g/kg BW, the serum histamine levels induced by compound 48/80 were reduced in a dose-dependent manner. We also investigated the effect of CDS on mast cell-dependent passive cutaneous anaphylaxis (PCA) activated by anti-dinitrophenyl (DNP) IgE antibody. CDS potently inhibited PCA when administered orally, topically, intraperitoneally or intradermally. However, it did not show inhibitory activity when administered intravenously. CDS dose-dependently inhibited the histamine release from the rat peritoneal mast cells (RPMC) by compound 48/80 and anti-DNP IgE. Moreover, the level of cAMP in RPMC, when CDS was added, significantly increased about 4-fold at 4 min compared with that of basal cells. These results indicate that CDS may possess strong antianaphylactic activity and also suggest the differential activity follwing administration routes may be caused by difference in bioavailability.

Solanum Lyratum Inhibits Anaphylactic Reaction and Suppresses the Expression of L- Histidine Decarboxylase mRNA

We investigated the effect of aqueous extract of Solanum lyratum Thunb. (Solanaceae) (SLAE) on anaphylactic reaction. The mast cell is widely thought to contribute to the acute changes associated with anaphylaxis. SLAE inhibited skin mast cells-mediated anaphylactic reaction activated by anti-dinitrophenyl (DNP) IgE. SLAE dose-dependently inhibited histamine release in mouse peritoneal mast cells activated by anti-DNP IgE or substance P. Substance P increased steady state levels of L-histidine decarboxylase (HDC) mRNA in mouse mastocytoma P-815 cells. Northern-blot analysis demonstrated that significantly reduced level of the mRNA of HDC was expressed in mast cells treated with SLAE, compared to that without SLAE. We conclude that SLAE directly affect IgE-mediated anaphylactic reaction and substance P-induced HDC mRNA over-expression.