Passive Cutaneous Anaphylaxis Test Research Articles

Allergenic response induced by Pterobothrium crassicolle (Cestoda: Trypanorhyncha) extracts in murine model.

The aim of this study was to determine the allergenic activity of components present in crude extracts of Pterobothrium crassicolle plerocerci (CPE) and blastocysts (CBE) obtained from Micropogonias furnieri in a murine model. Two groups of seven animals each received 50 µg of CPE or CBE on days 1, 35 and 120. Serum samples were tested by ELISA and Immunoblotting. Specific IgG and IgE levels were detected by ELISA, showing specific humoral responses for the primary immunization for both immunoglobulins and continuously growing titers for IgE. Positive Passive Cutaneous Anaphylaxis tests in rats sensitized with anti-CBE sera and tested by CBE, showed biologically, the allergenic activity of the extracts. The CPE and CBE showed some different recognition regions but both experimental groups recognized all regions of the extracts when tested for cross reactions, showing that CPE and CBE could share antigenic recognition sites.

Nonclinical safety evaluation of food colorant lac dye via systematic toxicity profiling with assessment of in vivo antigenic potential.

Lac dye is a natural colorant derived mainly from the insect Kerria lacca (Kerr) and has been used in food and beverage as a red-coloring additive. Despite its increasing use for human consumption as an alternative for allergy-associated cochineal, its toxicity profile remained incomplete to sufficiently assess its safety for the intended use. In this study, we evaluated systemic and genetic toxicity by performing acute and subacute oral toxicity studies in Sprague-Dawley (SD) rats using highly purified lac dye (LD) formulated in water and a battery of genotoxicity tests, respectively. To assess antigenic potentials, we carried out an in vivo passive cutaneous anaphylaxis test. A single dose of LD did not cause mortality at 5000mg/kg body weight (BW), setting oral LD50 of >5000mg/kg BW in SD rats. In the 90-day study, transient salivation without accompanying histopathological lesions in the salivary glands in 200 and 500mg/kg BW groups and red-purple pigmentation on the surface of femora and skulls in 500mg/kg groups were observed as nonadverse effects associated with LD, and no adverse effect was detected in all of the parameters examined, establishing a 500mg/kg BW as no-observed-adverse-effect-level (NOAEL). Furthermore, LD was not mutagenic nor clastogenic in the genotoxicity tests. When tested for antigenicity, LD did not induce anaphylactic skin responses as opposed to the positive reaction by ovalbumin, suggesting a lack of antigenicity. Taken together, these findings provide extended toxicity information on LD with direct evidence supporting the lack of antigenicity, providing essential guidance for its safe use in humans.

Vasicine alleviates 2,4-dinitrochlorobenzene-induced atopic dermatitis and passive cutaneous anaphylaxis in BALB/c mice

Atopic dermatitis (AD), a type of skin inflammation, is associated with immune response mediated by T-helper 2 (Th2) cells, and mast cells. Vasicine is an alkaloid isolated from Adhatoda vasica, a popular Ayurvedic herbal medicine used for treating inflammatory conditions. In the present study, the anti-AD effects of vasicine were evaluated on 2,4-dinitrochlorobenzene-induced AD-like skin lesions in BALB/c mice. The potential anti-allergic effects of vasicine were also assessed using the passive cutaneous anaphylaxis (PCA) test. The results showed that the oral administration of vasicine improved the severity of AD-like lesional skin by decreasing histopathological changes and restoring epidermal thickness. Vasicine also inhibited the infiltration of mast cells in the skin and reduced the levels of pro-Th2 and Th2 cytokines as well as immunoglobulin E in the serum. Finally, vasicine inhibited the expression of pro-Th2 and Th2 cytokines in skin tissues, indicating the therapeutic potential of vasicine for AD.

The quest for ingested peanut protein in human serum

There is mounting evidence that systemic uptake of food allergens is key to triggering anaphylaxis. However, direct proof for this theory is still lacking. The purpose of this study was to quantify the absorption and to determine the absorption kinetics of immunoreactive peanut protein in relation to the allergic response in human. Quantitative protein assays including mass spectrometry, dot blots and Western blotting were developed to determine the level of Ara h 2 absorption in human serum. The double monoclonal sandwich ELISA was applied to quantify absorbed Ara h 2 and 6, and the basophil histamine release assay and the human passive cutaneous anaphylaxis test were utilized to study the absorption kinetics of immunologically intact peanut proteins. The protein assays worked but were not sensitive enough to trace the minute amounts of absorbed Ara h 2 in human serum. The level of Ara h 6 in serum was found to be up to 0.2ng/mL, but Ara h 2 could not be detected with the ELISA. Both the in vivo and the in vitro methods were successful in demonstrating that: immunoreactive peanut protein was absorbed shortly after ingestion (≤5minutes); the peanut protein concentration peaks between 1 and 4hours; and peanut proteins can circulate for at least 48hours in the bloodstream. Ingested peanut protein is absorbed systemically and retains its immunoreactive capacity in human serum. However, the precise quantities and the implication for the elicitation of anaphylaxis remains to be elucidated.

Anti-allergic rhinitis effects of caffeoylquinic acids from the fruits of Xanthium strumarium in rodent animals via alleviating allergic and inflammatory reactions

The fruits of Xanthium strumarium L., Asteraceae, have been used for various diseases in Chinese folk medicine, including allergic rhinitis, tympanitis, arthritis, ozena etc. The current study was aimed to investigate the therapeutic effects of caffeoylquinic acids from fruits of X. strumarium on allergic rhinitis in animals. The toxicity test indicated that the caffeoylquinic acids have no obvious toxicity. By using HPLC assays combined with reference standards, ten caffeoylquinic acids were identified as the predominant constituents. Anti-allergic activities of the caffeoylquinic acids were evaluated using passive cutaneous anaphylaxis test and Schultz-Dale test; dimethylbenzene induced ear edema test was performed to evaluate its anti-inflammatory effect. Then, the allergic rhinitis model in rats was established to evaluate the therapeutic effects of the caffeoylquinic acids against allergic rhinitis with the following indexes: allergic rhinitis symptom scores, serum levels of pro-inflammatory cytokines, histopathological examination, and histamine release. Our study revealed that the caffeoylquinic acids showed obvious anti-allergic and anti-inflammatory properties, and its treatments were beneficial for ameliorating the nasal symptoms, decreasing pro-inflammatory cytokines, and inhibiting the releases of histamine. Collectively, the caffeoylquinic acids might be utilized as effective and safe disease therapeutic agents for allergic rhinitis.

Systemic and Local Anaphylaxis is Not Induced by Korean Red Ginseng Mixture in Guinea Pigs

Currently, injuries to customers due to health functional foods are annually increasing. To evaluate the antigenicity of Korean red ginseng mixture (KRGM), we tested for systemic anaphylactic shock and passive cutaneous anaphylaxis in guinea pigs. Based on a comparison of measured body weights, there were no changes in body weight for the KRGM treatment group compared with the control group. In the ovalbumin treated group, however, there was a statistically significant decrease in body weight. For the active systemic anaphylaxis test, after the induction, there were no symptoms that suggested anaphylactic shock in the control and KRGM treatment group. In the ovalbumin treated group, there were symptoms that suggested severe anaphylaxis, and those symptoms included restlessness, piloerection, tremor, rubbing or licking the nose, sneezing, coughing, hyperpnea, dyspnea, staggering gait, jumping, gasping and writhing, convulsion, side position and Cheyne-stokes respiration. All animals died within thirty minutes in the ovalbumin treated group. For the passive cutaneous anaphylaxis test in guinea pigs sensitized to KRGM, each anti-serum was diluted in a stepwise manner. This was followed by an intravenous injection of a mixture of KRGM and Evans blue. The results of the test showed that all the responses were negative in the control and the low-dose and high-dose administration groups. However, in the ovalbumin treated group, all the responses were positive. Based on the above results, there were no anaphylactic responses for up to 12 times the amount of human intake of KRGM in Hartley Guinea-pigs. The results suggest that KRGM is safe as measured by the systemic and local antigenicity in guinea pigs.

RhoA/ROCK Signaling Pathway Mediates Shuanghuanglian Injection-Induced Pseudo-allergic Reactions.

Background: Shuanghuanglian injection (SHLI) is a famous Chinese medicine used as an intravenous preparation for the treatment of acute respiratory tract infections. In the recent years, the immediate hypersensitivity reactions induced by SHLI have attracted broad attention. However, the mechanism involved in these reactions has not yet been elucidated. The present study aims to explore the characteristics of the immediate hypersensitivity reactions induced by SHLI and deciphers the role of the RhoA/ROCK signaling pathway in these reactions.Methods: SHLI-immunized mice or naive mice were intravenously injected (i.v.) with SHLI (600 mg/kg) once, and vascular leakage in the ears was evaluated. Passive cutaneous anaphylaxis test was conducted using sera collected from SHLI-immunized mice. Naive mice were administered (i.v.) with a single dose of 150, 300, or 600 mg/kg of SHLI, and vascular leakage, histamine release, and histopathological alterations in the ears, lungs, and intestines were tested. In vitro, human umbilical vein endothelial cell (HUVEC) monolayer was incubated with SHLI (0.05, 0.1, or 0.15 mg/mL), and the changes in endothelial permeability and cytoskeleton were observed. Western blot analysis was performed and ROCK inhibitor was employed to investigate the contribution of the RhoA/ROCK signaling pathway in SHLI-induced hypersensitivity reactions, both in HUVECs and in mice.Results: Our results indicate that SHLI was able to cause immediate dose-dependent vascular leakage, edema, and exudates in the ears, lungs, and intestines, and histamine release in mice. These were pseudo-allergic reactions, as SHLI-specific IgE was not elicited during sensitization. In addition, SHLI induced reorganization of actin cytoskeleton and disrupted the endothelial barrier. The administration of SHLI directly activated the RhoA/ROCK signaling pathway both in HUVECs and in the ears, lungs, and intestines of mice. Fasudil hydrochloride, a ROCK inhibitor, ameliorated the SHLI-induced hypersensitivity reactions in both endothelial cells and mice indicating its protective effect. SHLI-induced pseudo-allergic reactions were mediated by the activation of the RhoA/ROCK signaling pathway.Conclusion: This study presents a novel mechanism of SHLI-induced immediate hypersensitivity reactions and suggests a potential therapeutic strategy to prevent the associated adverse reactions.

Anti-allergic rhinitis effect of caffeoylxanthiazonoside isolated from fruits of Xanthium strumarium L. in rodent animals

The fruits of Xanthium strumarium L. (Asteraceae) have been used extensively in China for treatment of various diseases such as allergic rhinitis (AR), tympanitis, urticaria and arthritis or ozena. This study was designed to systemically investigate the effects of the caffeoylxanthiazonoside (CXT) isolated from fruits of X. strumarium on AR in rodent animals. Animals were orally administered with CXT. Anti-allergic activity of CXT was evaluated by passive cutaneous anaphylaxis test (PCA); acetic acid-induced writhing tests were used to evaluate the analgesic effects of CXT; acetic acid-induced vascular permeability tests were performed to evaluate anti-inflammatory effect of CXT. Then, the model AR in rats was established to evaluate the effects of CXT on AR with the following tests: the sneezing and nasal scratching frequencies, IgE level in serum, and histopathological examinations. Our results demonstrated that CXT had favorable anti-allergic, anti-inflammatory and analgesic effects. Additionally, we found that CXT was helpful to ameliorate the nasal symptoms and to down-regulate IgE levels in AR rats. Thus, we suggested that CXT can be treated as a candidate for treating AR.

Experimental study on antinociceptive and anti-allergy effects of patchouli oil

Pogostemon cablin (Blanco) Benth. (Lamiaceae), which is named ‘Guang-Huo-Xiang’ in Chinese, has various therapeutic functions to remove dampness, relieve summer-heat and exterior syndrome, stop vomiting, and stimulate appetite. Patchouli oil is the essential oil of Pogostemonis herba. This study is aimed to investigate the antinociceptive and anti-allergy activities of patchouli oil. The antinociceptive activity of patchouli oil (20, 40, and 80 mg/kg) was evaluated using the acetic acid-induced writhing test (NIH mice) and the hot-plate test (NIH mice), and those for anti-allergy activity were tested using Schultz–Dale reaction, the passive cutaneous anaphylaxis (PCA) test, and delayed-type hypersensitivity (DTH) test. All doses of patchouli oil could significantly (p<0.05) extend writhing latent period and moderately decrease writhing frequency. In the hot-plate test, patchouli oil treatment significantly (p<0.01) increased the latency period at all three doses at 30, 60, and 90-minute time intervals compared with the control group. Patchouli oil could remarkably (p<0.01) inhibit the contraction induced by the Schultz–Dale reaction. Moreover, patchouli oil at all doses significantly (p<0.01) inhibited the PCA reaction induced by Sprague–Dawley rat anti-ovalbumin serum and the DTH reaction induced by the 2,4-dinitrochlorobenzene when comparing with those of the control group. The findings suggested that patchouli oil may provide additional evidence for its potential uses as antinociceptive and anti-allergy agents.

Experimental Study of Antigenicity of Sweet Bee Venom in Guinea Pigs

Objectives: This study was performed to examine the antigenic potential of pure melittin (Sweet Bee Venom - SBV) extracted from the bee venom by utilizing protein isolation method of gel filtration. Methods: All experiments were conducted at Biotoxtech (Chungwon, Korea), authorized a non-clinical studies institution, under the regulations of Good Laboratory Practice (GLP). Antigenic potential of SBV was examined by active systemic anaphylaxis (ASA) and passive cutaneous anaphylaxis (PCA) in guinea pigs. SBV was subcutaneously administered at 0.07 and 0.28mg/kg and also as a suspension with adjuvant (Freund`s complete adjuvant: FCA). Ovalbumin (OVA) as a suspension with adjuvant was used to induce positive control response (-FCA). Results: 1. In the ASA test, experimental groups showed some symptoms of anaphylaxis like piloerection, hyperpnea and staggering gait. 2. In the PCA test, low dosage group did not show any antibody responses, whereas high dosage group showed positive responses. 3. In the weight measurement and clinical observation, experimental groups didn`t show any significant changes compared with control group. 4. In the autopsy of body, the abnormalities of lung were detected in the corpse. This means that the cause of death may induced anaphylactic shock. Conclusions: Above findings suggested that SBV had antigenic potential in guinea pig. Further studies on the subject should be conducted to yield more concrete evidences.

Preparation, Characterization and Related &lt;i&gt;in Vivo&lt;/i&gt; Release, Safety and Toxicity Studies of Long Acting Lanreotide Microspheres

The goal of this project was to prepare long-acting lanreotide acetate poly(lactic-co-glycolic acid) (PLGA) microspheres and to analyze the in vivo and in vitro release, safety and toxicology of these preparations. Long-acting lanreotide acetate PLGA microspheres that exhibited a 5-week slow-release period were prepared by a multiple-emulsion solvent evaporation method. Physical characterization, as well as the analysis of the in vivo and in vitro release, safety, acute toxicity and chronic toxicity of the lanreotide microspheres, were conducted in animal models in rats, guinea pigs, rabbits and beagle dogs. The lanreotide acetate PLGA microspheres prepared by multiple-emulsion solvent evaporation had smooth surfaces, uniform particle size and stable lanreotide loading. In vivo and in vitro experiments showed that the lanreotide acetate PLGA microspheres could continuously release lanreotide for 5 weeks. The safety of these long acting lanreotide microspheres was good in the following animal models: active systemic anaphylaxis test in guinea pigs, passive cutaneous anaphylaxis test in rats, hemolytic test in rabbits, local skin irritation test after subcutaneous administration in rabbits and muscle stimulation test in rabbits. Furthermore, no significant acute toxicity or chronic toxicity was observed after administration of lanreotide acetate PLGA microspheres in beagle dogs at dosages up to 22 mg/kg. The lanreotide acetate PLGA microspheres that were prepared in this study exhibited beneficial characteristics in apparent property and structural stability, as well as in release trends in vivo and in vitro.

기니픽을 이용한 Sweet Bee Venom의 항원성 평가

Objectives: This study was performed to examine the antigenic potential of pure melittin (Sweet Bee Venom - SBV) extracted from the bee venom by utilizing protein isolation method of gel filtration. Methods: All experiments were conducted at Biotoxtech (Chungwon, Korea), authorized a non-clinical studies institution, under the regulations of Good Laboratory Practice (GLP). Antigenic potential of SBV was examined by active systemic anaphylaxis (ASA) and passive cutaneous anaphylaxis (PCA) in guinea pigs. SBV was subcutaneously administered at 0.07 and 0.28mg/kg and also as a suspension with adjuvant (Freund`s complete adjuvant: FCA). Ovalbumin (OVA) as a suspension with adjuvant was used to induce positive control response (-FCA). Results: 1. In the ASA test, experimental groups showed some symptoms of anaphylaxis like piloerection, hyperpnea and staggering gait. 2. In the PCA test, low dosage group did not show any antibody responses, whereas high dosage group showed positive responses. 3. In the weight measurement and clinical observation, experimental groups didn`t show any significant changes compared with control group. 4. In the autopsy of body, the abnormalities of lung were detected in the corpse. This means that the cause of death may induced anaphylactic shock. Conclusions: Above findings suggested that SBV had antigenic potential in guinea pig. Further studies on the subject should be conducted to yield more concrete evidences.

Alpha-momorcharin possessing high immunogenicity, immunotoxicity and hepatotoxicity in SD rats

Momordica charantia L., a genus of Momordica Linn. of the family Cucurbitaceae, commonly known as bitter melon, has been widely planted in China, Southeast Asia, Turkey and other areas, and has been used as a medicine for a long time. Alpha-momorcharin (α-MMC) extracted and purified from bitter melon seeds has significant anti-tumor and anti-virus effects, and has potential toxicity as well, especially when taken overdose. However, up to date studies on its safety evaluation are still insufficient. Aims of the studyThe immunogenicity, immunotoxicity and general toxicity of α-MMC were investigated in rats and guinea-pigs, and the potential toxic effects of the agent on the body were also examined. Materials and methodsThe major ribosome-inactivating protein was isolated by column chromatographies from the protein extracted from bitter melon seeds, and was verified as α-MMC. After rats were immunized by α-MMC, titers of specific antibody to α-MMC in immunized rats serum were detected by indirect ELISA. Guinea-pigs and rats immunized with α-MMC were used to evaluate the active systemic anaphylaxis and passive cutaneous anaphylaxis induced by α-MMC relatively. α-MMC of 6.25mg/kg, 2.08mg/kg and 0.70mg/kg was administered to rats every 2 days. Five weeks later, animals were sacrificed, and then, biochemical examination, analysis of bone marrow and peripheral blood cells, and histopathologic examination were performed. ResultsThe ribosome-inactivating protein isolated and purified from bitter melon seeds was identified as α-MMC. It induced high titer (1:46.4) of specific IgG and high positive results of the active systemic anaphylaxis and passive cutaneous anaphylaxis tests in animals. With the time of the α-MMC administration increasing, the body weights of the animals administered with α-MMC of 6.25mg/kg decreased significantly, and point necrosis was also observed in liver cells, along with abnormal findings in serum chemistry, hematology and bone marrow histopathology test. The toxic effect lessened with the decrease of the dose of α-MMC and further reduced after the convalescence stage. ConclusionsThe results of the study show that α-MMC has high immunogenicity and immunotoxicity, and can cause obvious organic liver lesion.

Airway responsiveness in an allergic rabbit model

Background Animal models of allergy and bronchial hyperresponsiveness (BHR) are useful in researching pulmonary diseases and evaluating drug effects on the airways. Neonatally immunised rabbits exhibit several features of asthma as adults, including early and late airway responses following antigen challenge, oedema and inflammatory cell infiltration into the lung, BHR to inhaled histamine and methacholine (compared with naïve rabbits) and exacerbations of BHR following antigen challenge. Therefore this model can be used to investigate the underlying mechanisms of BHR and for the preclinical evaluation of new drugs for the treatment of asthma. Aim To describe the characteristics of airway responses in a rabbit model of allergic inflammation and to evaluate the relationship between skin test reactivity to antigen, airway inflammation and BHR. Methods New Zealand White rabbits were neonatally immunised against Alternaria tenius. At 3 months of age, airway responsiveness was measured to aerosolised histamine, methacholine or allergen. Bronchoalveolar lavage (BAL) was performed and cell counts recorded. Direct skin tests were performed to assess skin reactivity to allergen and passive cutaneous anaphylaxis (PCA) tests were performed to determine titres of circulating IgE. Results In a population of allergic rabbits, allergen challenge induced a significant bronchoconstriction, airway inflammation and BHR. Skin test responsiveness to allergen did not correlate with various indices of pulmonary mechanics e.g. baseline sensitivity to methacholine and histamine, or allergen-induced BHR. In contrast, skin test responsiveness did predict airway inflammation as assessed by measurements of eosinophil recruitment to the lung. Conclusion The allergic rabbit is a useful model to further our understanding of allergic diseases. Recording lung function using a minimally invasive procedure allows repeated measurements to be made in rabbits longitudinally, and each animal may thus be used as its own control. Our observations do not support the use of skin responsiveness to allergen as a predictor of airway sensitivity as we observed no correlation between skin sensitivity and airway responsiveness to inhaled histamine, methacholine or allergen. However, skin reactivity did predict airway inflammation as assessed by measurements of eosinophil recruitment to the lung. Our results also further highlight the likely dissociation between airway inflammation and BHR.

Effect of traditional Chinese medicine injections on type I allergy

To investigate allergic reactions of traditional Chinese medicine (TCM) injections, and to determine the contents of serum IgE and histamine in sensitized animal. The correlation between the preceding contents in serum and allergic reactions may be found, thus offering experimental evidences for advancing the accuracy of anticipation by type I allergy. We carried out passive cutaneous anaphylaxis (PCA) tests,active systemic anaphylaxis (ASA) tests and anaphylactoid reactions using three TCM injections, and determined the contents of serum OVA-sIgE, total serum IgE and histamine in sensitized animals by ELISA method. The results of PCA test were negative, and there was no significant difference for total serum IgE level between experimental group and normal saline group. In the study of adjuvant effect in TCM injections + OVA (at the dose level that doesn't cause allergic reactions), the PCA results of SHL and YXC were positive and there was a increase in content of serum OVA-sIgE, while the PCA result of QKL was negative with a unobvious increase in the content of serum OVA-sIgE. The content of total serum IgE wasn't remarkably increased in each group and the results of ASA test were all positive. Three injections all caused anaphylactoid symptoms in guinea pigs in different doses or injection speed and the response intensity was found to be dosage and injection speed dependant. Furthermore, there was no significant difference for the content of total serum IgE in each group, whereas serum histamine concentration in every experimental group was markedly higher than normal saline group. SHL and YXC increase the sensitivity of guinea pigs on OVA, and three TCM injections can cause allergic reactions in guinea pig. Allergic reactions of three TCM injections are correlated with specific IgE antibodies and histamine contents.

Study of immunogenicity of chlorogenic acid-BSA with different number of CGA

To investigate the immunogenicity of chlorogenic acid-BSA(CGA-BSA) and the foundation for the allergization of CGA-BSA. The CGA-BSA with different number of CGA was synthesized to allergize the BALB/c mice and SD rats to get antiserum. The level of IgE and histamine was analyzed by ELISA, and the passive cutaneous anaphylaxis (PCA) test was carried to analyze the antibody titer. When the coupling rate of CGA-BSA is 20, the highest IgE and histamine level in BALB/c mice and the antibody titer in PCA are the highest. It has better immunogenicity when the coupling rate of CGA-BSA is between 10 to 20, while the CGA-BSA 20 with the best immunogenicity.

Montelukast treatment (cysteinyl leukotriene receptor antagonist) in a model of food allergy: modifications in lymphatic cell population from rectal mucosa

The aim is to determine immunopathological modifications in rectal mucosa from rabbits after local challenge in ovalbumin (OVA) sensitized animals previously treated with montelukast. thirty two rabbits divided into four groups: G1: normal; G2: subcutaneously OVA sensitized; G3: sensitized, locally OVA challenged and sampled 4 hours after challenge; and G4: sensitized, locally OVA challenged and treated 4 hours before challenge with montelukast (0.15 mg/kg). Specific anti-OVA IgE levels were evaluated by passive cutaneous anaphylaxis test (PCA). In each group 200 high microscopical power fields (HPF) were counted. Results were expressed as arithmetic mean and SE. Anti -CD4, CD5, micro chain monoclonal antibodies were used. Avidin biotin horseradish peroxidase system was used. CD 4: G1: 8.3 +/- 0.06; G2: 13.4 +/- 0.08, G3: 8.25 +/- 0.06, G4: 11.8 +/- 0.02. CD 5: G1: 7.3 +/- 0.05; G2: 9.4 +/- 0.05, G3: 11.3 +/- 0.06, G4: 8.1 +/- 0.06. mu chain: G1: 10.4 +/- 0.06; G2: 3.8 +/- 0.02, G3: 6.0 +/- 0.10, G4: 2.2 +/- 0.10. In all cases, experimental groups (G3 vs. G4) presented statistical significant differences (p < 0.05). CD4+, CD5+ cells and mu chain+ decrease in experimental group (G4), probably due to lymphocyte migration inhibition to challenged mucosa. mu chain+ cell decrease could be based on B cell activation and expression of different surface immunoglobulins. Cells expressing mu chain decreased in G2 and G3 likely due to activation of B cells and subsequent expression of other immunoglobulin chains in cell surface. We conclude that obtained data are important to elucidate immunopathology of local anaphylactic reaction in rectal mucosa from systemic sensitized animals after treatment with montelukast.

Isolation and purification of anaphylactic antibody from guinea pig and preliminary research on its role in passive cutaneous anaphylaxis test (PCA)

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. Key words: Guinea pig; Type Ⅰ anaphylaxis; Passive cutaneous anaphylaxis; IgE; IgG

Evaluation of the allergenicity of goat milk, cow milk, and their lactosera in a guinea pig model

The object of this study was to determine the allergenicity of goat milk (GM) and cow milk (CM) and that of their respective lactosera (GML and CML), by in vivo and in vitro assays. Two systemic tests for anaphylaxis were carried out in guinea pigs, the animals being sensitized orally with the 2 types of milk and lactosera. Sera were taken from the orbital sinus of the experimental animals at 0 and 22 d of the experiment to perform the serological study and the passive cutaneous anaphylaxis test. For the latter, the guinea pigs were sensitized passively with antibodies against the 4 antigen solutions. Enzyme-linked immunosorbent assay and Western blot were used to determine the specific antibodies of the isotypes immunoglobulin G1 and immunoglobulin G(Fc) developed against the same 4 antigen solutions. From these anaphylaxis and antibody-production tests, it was concluded that GM is hypoallergenic when compared with CM. The lactosera produced more closely grouped results, with values always below those of the corresponding milk. None of the proteins in the 4 immunizing solutions were identified as being their main allergen. These results show the hypoallergenicity of GM versus CM, and also that both casein and lactoserum proteins may be responsible for allergy in each case. To analyze the possibility of producing an innocuous food for those allergic to milk proteins, it would be of interest to identify the epitope(s) responsible for such allergenicity.

감마선 조사에 의한 분유 단백질의 항원성 저감화

본 연구는 안전성 확보차원에서 식품분야에 활용성이 증가하고 있는 감마선 조사처리가 유제품의 항원성 저감에도 효과가 있음을 보이기 위해 수행되었다. 모세관전기영동 결과 <TEX>${\alpha}_{S1}$</TEX>-케이신과 <TEX>${\beta}_{A1}$</TEX>-케이신이 <TEX>${\alpha}_{S0}$</TEX>-케이신과 <TEX>${\beta}_{A2}$</TEX>-케이신보다 상대적으로 많이 분해되었음을 확인하였다. cELISA를 이용한 in vitro 시험 결과와 기니픽을 이용한 수동피부아나필랙시스(Passive cutaneous anaphylaxis) 반응을 통해 10 kGy의 감마선 조사에서 뚜렷한 항원성 저감을 확인하였다. 이상의 결과는 감마선 조사가 우유 단백질의 항원, 특히 <TEX>${\alpha}_{S1}$</TEX>-케이신과 <TEX>${\beta}_{A1}$</TEX>-케이신의 구조를 변화시켜 항원성을 감소시키는 것으로 판단되며, 따라서 감마선 조사는 알레르기 유발 식품의 항원성 감소에 유용한 기술로 생각된다. The aim of this work was to examine the reduction in antigenicity of milk proteins in powdered milk by gamma irradiation which is increasingly used for food safety. Skim milk powder samples were exposed to irradiation doses of 1, 5, and 10 kGy. A greater reduction of <TEX>${\alpha}_{S1}$</TEX>-casein and <TEX>${\beta}_{A1}$</TEX>-casein was found than <TEX>${\alpha}_{S0}$</TEX>-casein and <TEX>${\beta}_{A2}$</TEX>-casein by capillary electrophoresis. Competitive indirect ELISA and passive cutaneous anaphylaxis tests using guinea pigs showed a reduction in antigenicity of powdered milk by 10kGy gamma irradiation. These results indicated that gamma irradiation reduce allergenicity of milk proteins by structural changes of <TEX>${\alpha}_{S1}$</TEX>-casein and <TEX>${\beta}_{A1}$</TEX>-casein, and can be useful for dairy products.