Tryptophan Negatively Regulates IgE-mediated Mast Cell Activation / 체질인류학회지

Mast cells are major immune cells in allergy to secrete allergic mediators by a degranulation process and make and secrete inflammatory lipids and cytokines in response to antigen stimulation. An amino acid tryptophan regulates immune functions. Tryptophan ameliorates inflammatory colitis in which mast cells are engaged. However, its effects on mast cells remain to be solved. We investigated the effect of tryptophan on IgE-mediated allergic responses in the mast cells and mice. IgE-mediated passive cutaneous anaphylaxis (PCA) in mice were examined. Also IgE-mediated mast cell activation responses such as degranulation of stored granules and secretion of inflammatory lipid LTB₄ and cytokines (TNF-α and IL-4) were measured. Intraperitoneal administration of tryptophan suppressed PCA in mice. Also, in the cellular level tryptophan inhibited IgE-mediated mast cell activation such as IgE-mediated degranulation and the production of LTB₄. Also, it inhibited production of inflammatory cytokines TNF-α and IL-4. In summary, tryptophan suppressed IgE-mediated allergic activation in vivo and in vitro. Tryptophan supplementation is beneficial for IgE-mediated allergy.

Study on allergy of Astragaloside injection / 药物评价研究

Objective To evaluate the hypersusceptibility of Astragaloside injection on animal,and provide reference for clinical use with active systemic anaphylaxis (ASA),passive cutaneous anaphylaxis (PCA) and determination of serum sample titer.Methods ASA:Guinea pigs was ip with 0.4,1.6 mg/kg Astragaloside injection five times every other day.On the eleventh day after the last administration,the test substance was quickly injected to fore limb vein,and animal allergy symptoms were observed within 30 min.PCA:Astragaloside injection was ip injected to rats five times every other day and antiserum was collected.The antiserum was appropriately diluted,and sc injected to another group rats for passive sensitization.About 48 hours later,Astragaloside was quickly iv to rats,and the skin allergy was observed.Meanwhile,the antibody titer of the antiserum was determined.Results ASA:Astragaloside injection of 0.4,1.6 mg/kg in guinea pigs did not show any allergic reaction,that is,ASA was negative;PCA:Astragaloside injection of 0.5,2.0 mg/kg in rats did not show any allergic reaction,and Astragaloside specific antibodies were not determined in serum samples.That is,PCA was negative.Conclusion The results of ASA and PCA were negative in the experimental dose,and there was no specific antibody against Astragaloside in the serum prepared by PCA,which indicated that the possibility of hypersensitivity reaction was weak in clinical use.

Study on allergy of Astragaloside injection / 药物评价研究

Objective To evaluate the hypersusceptibility of Astragaloside injection on animal,and provide reference for clinical use with active systemic anaphylaxis (ASA),passive cutaneous anaphylaxis (PCA) and determination of serum sample titer.Methods ASA:Guinea pigs was ip with 0.4,1.6 mg/kg Astragaloside injection five times every other day.On the eleventh day after the last administration,the test substance was quickly injected to fore limb vein,and animal allergy symptoms were observed within 30 min.PCA:Astragaloside injection was ip injected to rats five times every other day and antiserum was collected.The antiserum was appropriately diluted,and sc injected to another group rats for passive sensitization.About 48 hours later,Astragaloside was quickly iv to rats,and the skin allergy was observed.Meanwhile,the antibody titer of the antiserum was determined.Results ASA:Astragaloside injection of 0.4,1.6 mg/kg in guinea pigs did not show any allergic reaction,that is,ASA was negative;PCA:Astragaloside injection of 0.5,2.0 mg/kg in rats did not show any allergic reaction,and Astragaloside specific antibodies were not determined in serum samples.That is,PCA was negative.Conclusion The results of ASA and PCA were negative in the experimental dose,and there was no specific antibody against Astragaloside in the serum prepared by PCA,which indicated that the possibility of hypersensitivity reaction was weak in clinical use.

Optimization of parameters of passive cutaneous anaphylaxis in rats / 中国药理学与毒理学杂志

OBJECTIVE To optimize the parameters of passive cutaneous anaphylaxis(PCA)in rats immunized by ovalbumin(OVA). METHODS 1-2 month-old Sprague-Dawley rats were immu?nized by ip injection of OVA(0.2,1.0 and 5.0 mg per rat)mixed with complete Freund′s adjuvant once every other day 3 times. Serum was collected on the 12th-16th days after final immunization. Then the rats were intracutaneously injected with sensitized serum and then stimulated by iv injection of the same dose of OVA mixed with Evans blue after a latent period of 0.5,1.5,3,6,12,24,36,48 and 60 h. Finally,the diameters of blue spots in the skin were measured at stimulation. RESULTS Serum total-IgE(T-IgE)and OVA-specific IgE(sIgE)levels increased significantly and reached the peak on the 3rd-7th days and 12th-16th days after final immunization,respectively. There was no correlation between the serum T-IgE level and OVA-sIgE level when the rats were immunized with OVA at OVA 0.2-5.0 mg per rat. The rats experienced PCA after injection of OVA 1.0 and 5.0 mg per rat. Diameters of blue spots in the skin reached the maximum value after rats were sensitized for 0.5-3 h. Moreover,the shape,color and size of blue spots were better 30-60 min after stimulation. CONCLUSION Optimized PCA is as follows:1-2 month-old rats are immunized on the 1st,3rd and 5th days by ip injection of OVA 1.0-5.0 mg. The immunizing serum is collected at 12-16 d after final immunization. The rats are stimulated by OVA and Evans blue after a latent period of 0.5-3 h. Diameters of blue spots in rats′ skin are then measured 30-60 min after stimulation.

Antibody to FcεRIα Suppresses Immunoglobulin E Binding to High-Affinity Receptor I in Allergic Inflammation

PURPOSE: High-affinity receptor I (FcεRI) on mast cells and basophils plays a key role in the immunoglobulin E (IgE)-mediated type I hypersensitivity mediated by allergen cross-linking of the specific IgE-FcεRI complex. Thus, prevention of IgE binding to FcεRI on these cells is an effective therapy for allergic disease. We have developed a strategy to disrupt IgE binding to FcεRI using an antibody targeting FcεRIα. MATERIALS AND METHODS: Fab fragment antibodies, which lack the Fc domain, with high affinity and specificity for FcεRIα and effective inhibitory activity against IgE-FcεRI binding were screened. IgE-induced histamine, β-hexosaminidase and Ca2+ release in basophils were determined by ELISA. A B6.Cg-Fcer1a(tm1Knt) Tg(FCER1A)1Bhk/J mouse model of passive cutaneous anaphylaxis (PCA) was used to examine the inhibitory effect of NPB311 on allergic skin inflammation. RESULTS: NPB311 exhibited high affinity to human FcεRIα (KD=4 nM) and inhibited histamine, β-hexosaminidase and Ca2+ release in a concentration-dependent manner in hFcεRI-expressing cells. In hFcεRIα-expressing mice, dye leakage was higher in the PCA group than in controls, but decreased after NPB311 treatment. NPB311 could form a complex with FcεRIα and inhibit the release of inflammation mediators. CONCLUSION: Our approach for producing anti-FcεRIα Fab fragment antibody NPB311 may enable clinical application to a therapeutic pathway in IgE/FcεRI-mediated diseases.

Antibody to FcεRIα Suppresses Immunoglobulin E Binding to High-Affinity Receptor I in Allergic Inflammation

PURPOSE: High-affinity receptor I (FcεRI) on mast cells and basophils plays a key role in the immunoglobulin E (IgE)-mediated type I hypersensitivity mediated by allergen cross-linking of the specific IgE-FcεRI complex. Thus, prevention of IgE binding to FcεRI on these cells is an effective therapy for allergic disease. We have developed a strategy to disrupt IgE binding to FcεRI using an antibody targeting FcεRIα. MATERIALS AND METHODS: Fab fragment antibodies, which lack the Fc domain, with high affinity and specificity for FcεRIα and effective inhibitory activity against IgE-FcεRI binding were screened. IgE-induced histamine, β-hexosaminidase and Ca2+ release in basophils were determined by ELISA. A B6.Cg-Fcer1a(tm1Knt) Tg(FCER1A)1Bhk/J mouse model of passive cutaneous anaphylaxis (PCA) was used to examine the inhibitory effect of NPB311 on allergic skin inflammation. RESULTS: NPB311 exhibited high affinity to human FcεRIα (KD=4 nM) and inhibited histamine, β-hexosaminidase and Ca2+ release in a concentration-dependent manner in hFcεRI-expressing cells. In hFcεRIα-expressing mice, dye leakage was higher in the PCA group than in controls, but decreased after NPB311 treatment. NPB311 could form a complex with FcεRIα and inhibit the release of inflammation mediators. CONCLUSION: Our approach for producing anti-FcεRIα Fab fragment antibody NPB311 may enable clinical application to a therapeutic pathway in IgE/FcεRI-mediated diseases.

Effect of Traditional Chinese Medicine Injections on Allergic Reactions / 世界科学技术-中医药现代化

This study was aimed to investigate allergic reactions of traditional Chinese medicine (TCM) injections, and to determine contents of serum IgE in sensitized animals. The correlation between preceding contents in serum and allergic reactions may be found, in order to offer experimental evidences for advancing the accuracy of anticipa-tion by allergic reactions. Passive cutaneous anaphylaxis (PCA) tests and anaphylactoid reactions were used in a vari-ety of TCM injections. ELISA method was used to determine the content of total serum IgE in sensitized animals. The PCA results of SHL, QKL, YXC, XST, GGS and CHN were negative. The PCA result of CWJ was positive. There was no significant difference for total serum IgE level between the experimental group and the normal saline group in the group of SHL, QKL, YXC, XST, GGS, CHN and CWJ. All TCM injections caused anaphylactoid symp-toms in guinea pigs. It was concluded that all TCM injections can cause allergic reactions in guinea pig. And the al-lergic reactions of TCM injections were not correlated with serum total IgE.

Immunotoxicity study of Ginenoside Compound K Injection / 国际中医中药杂志

Objective Evaluate the immunotoxicity of Ginenoside Compound K Injection.Methods Active Systemic Anaphylaxis (ASA)tests and Passive Cutaneous Anaphylaxis (PCA)tests were used to evaluate Ginenoside Compound K Injection.Results In the ASA tests,positive control group showed pole-strength anapbylaxis,both the high-dose group and the low-dose group of Ginenoside Compound K Injection didn't produce allergic reaction and the body weights of all groups showed no significant differences.In the PCA tests,all rats of positive control group caused blue spots with their diameters bigger than 5 mm (diameters on the left side of blue spots was (10.1± 3.34) mm and diameters on the right side of blue spots was(7.57± 1.94)mm.Serum IgE was significantly increased.While both high dose and low dose of Ginenoside Compound K Injection group didn't show blue spots with their diameters greater than 5 mm and their IgE levels showed no significant differences compared with negative control group.Conclusion Ginenoside Compound K Injection showed no immunotoxicity under these experimental conditions.

Isolation and purification of anaphylactic antibody from guinea pig and preliminary research on its role in passive cutaneous anaphylaxis test (PCA) / 中华微生物学和免疫学杂志

Objective To establish guinea pig model of type Ⅰ anaphylaxis, isolate the anaphy-lactic antibody(IgE and IgG) preliminarily from sera of sensitized guinea pigs, and investigate its functional characteristics. Methods Animal model was established by sensitizing guinea pigs with OVA and Al(OH)_3, level of antibody was determined by ELISA, IgE and IgG in sera were preliminarily isolated through saturated ammonium sulphate precipitate and affinity chromatograph of Protein A. A continuous passive cutaneous ana-phylaxis test (PCA) was performed by sensitizing guinea pigs individually with IgE and IgG and then challenging at different time , and the variation of blue spots in skin were observed after challenge . Results Concentration of IgE in model group and control group were 719.3750 ng/ml and 2.5250 ng/ml, the optical density of IgG in model group and control group were 0.9921 and 0.0174, the level of two antibodies in model group were significantly higher than that of control group (P<0.05). In 9 d continuous PCA test, the blue spot induced by IgE in skin lasted for 9 days and appeared the largest when challenged at day 5. The diameter of blue spot induced by IgG was the largest when challenged at day 2 and then decreased fast. Con-clusion Anaphylactic antibodies were successfully preliminarily isolated from sera of sensitizing guinea pig, both IgE and IgG play roles in type Ⅰ anaphylaxis of guinea pig, the hypersensitive reaction induced by IgG is fast and short than that induced by IgE, and IgG may become an important surrogate marker in immunotoxic-ity evaluation(type Ⅰ anaphylaxis)of vaccine.

Efecto de Plectranthus amboinicus (Lour.) Spreng. tabletas sobre la anafilaxia pasiva cutánea, transmisión histaminérgica y adrenérgica / Effect of Plectranthus amboinicus (Lour.) Spreng tablets on the passive cutaneous anaphylaxis, histaminergic and adrenergic transmission.

Plectranthus amboinicus (Lour.) Spreng., más conocido en nuestro país como orégano francés, es una planta a la que se le confieren una gran cantidad de propiedades terapéuticas para el tratamiento de diferentes enfermedades dentro de las que se encuentra el asma bronquial. En el presente trabajo se realizó la evaluación de las tabletas 100 mg de Plectranthus amboinicus sobre la anafilaxia pasiva cutánea y la transmisión adrenérgica e histaminérgica. Como resultado final de este trabajo pudimos comprobar que Plectranthus amboinicus tabletas de 100 mg inhibe la anafilaxia pasiva cutánea, potencia la transmisión adrenérgica e inhibe los efectos de la histamina cuando esta interactúa con los receptores H1 presentes a nivel intestinal. Podemos concluir que las tabletas de 100 mg de Plectranthus amboinicus podrían emplearse en el tratamiento de enfermedades alérgicas tipo I y con esto justificar el uso popular de la planta en el asma bronquial.

Plectranthus amboinicus (Lour.) Spreng, well-known in our country as French origan, is a plant with many therapeutical properties for the treatment of different diseases, including bronchial asthma. In the present paper, the effect of Plectranthus amboinicus tablets (100 mg) on the passive cutaneous anaphylaxis and on the adrenergic and histaminergic transmission was evaluated. As a final result of this paper, it was possible to confirm that these tablets inhibit the passive cutaneous anaphylaxis, potentiates the adrenergic transmission, and inhibits the effects of histamine when it interacts with H1 receptors present at the intestinal level. It was concluded that the Plectranthus amboinicus tablets 100 mg could be used in the type I treatment of allergic diseases, and thus justify the popular use of this plant for bronchial asthma.

Evaluation of tryptase measurement in diagnosing allergic diseases / 中华检验医学杂志

Objective To evaluate the use of tryptase in diagnosing allergic diseases.Methods Serum tryptase was measured by Unicap 100 in 35 patients with different types and states of allergic diseases, and 30 healthy volunteers as controls ;Serum tryptases were compared with IgE level.Results Among the 35 allergic patients, 18 cases were with chronic urticaria(4.74?2.54)?g/L; 10 cases had past histories of anaphylaxis shocks(4.86?2.55)?g/L ;7 cases were in acute phase of anaphylaxis(17.4?7.87)?g/L ;30 healthy volunteers(4.36?1.67)?g/L. A statistically significant increase in tryptase concentration was found in patients with acute phase of anaphylaxis compared with other groups.No correlation was found between tryptase concentration and total IgE level.Conclusions Measurement of Serum tryptase level could be an additional tool for diagnosis of anaphylaxis but not chronic urticaria.

Inhibitory Effect of Danggui Decoction on Passive Cutaneous Anaphylaxis in Mice / 广州中医药大学学报

【Objective】To explore the mechanism of Danggui Decoction (DD) in counteracting allergic reaction bymeans of passive cutaneous anaphylaxis (PGA) test.【Methods】Among fifty NEH mice, ten were used to prepareantiserum and the rest forty were randomized into 4 groups: normal control group (A), clarityne group (B), low-doseDD group (C) and high-dose DD group (D) . PGA was induced by intraperitoneal injection of antiserum in the fourgroups and the mice were treated with gastric infusion simultaneously. After the last feeding, antigen attack wasconducted. Inhibitory rate of PCA in four groups were measured by colorimetric analysis after treatment. 【Results】High-and low-dose DD and clarityne could inhibit PCA in mice (P0.05).【Conclusion】The anti-allergic effect of DD may be associated with the inhibition of type Ⅰ allergic reaction.

Regulation of IgE antibody production in mice Ⅲ.IgE antibody response to hapten-carrier conjugate / 中国免疫学杂志

The anti-DNP and anti-OA IgE antibody responses of six strains of mice immunizedwith DNP-OA conjugate in presence of A1(OH)_3 adjuvant was investigated.Signifi-cant differences of the magnitude of serum IgE were found in anti-hapten and anti-carrier responses among strains.The primary and secondary anti-DNP and anti-OAIgE antibodies were elicited in NIH,BALB/c and DBA/2 mice after immunizationwith 1?g of DNP-OA.Among them NIH mice was the highest responder,whereasLACA,C3H and B6D2F1 mice were poor or nonresponders.In comparison the IgEresponse in BALB/c mice immunized with 1?g and 10?g of DNP-OA,the former de-monstrated a latency of primary response,but two groups had a very similar PCAtiters after 4 weeks of immunization.On the other hand,LACA mice gave a dose de-pendent IgE response.Profound primary and secondary IgE antibody responses can beonly seen in mice injected with 10?g of DNP-OA and PCA titer of anti-DNP washigher than that of anti-OA IgE antibody.However,no difference of IgE antibodylevels in NIH mice can be found between groups immunized with different doses.DNP_(4.7)-OA primed mice gave higher IgE response than that of mice immunized withDNP_3-OA.A prevailing anti-hapten IgE response was observed when higher dose ofDNP-OA conjugates was injected.