Effect of Kumhwang-San on anaphylactic reaction in a murine model

We investigated the effect of Kumhwang-San on anaphylactic reaction. Kumhwang-San dose-dependently inhibited mast cell-mediated systemic anaphylactic reaction induced by compound 48/80. Kumhwang-San significantly inhibited plasma histamine levels induced by compound 48/80. Kumhwang-San dose-dependently inhibited histamine release from the rat peritoneal mast cells activated by compound 48/80 or substance P. The level of cAMP in rat peritoneal mast cells, when Kumhwang-San was added, transiently and significantly increased about 3-fold compared with that of basal cells. These results suggest that Kumhwang-San will be beneficial in the treatment of anaphylactic reaction.     

Sensitizing capacity and residual allergenicity of hydrolyzed cow's milk formulae: results from a murine model

BACKGROUND: Cow's milk allergy is the most common cause of clinically relevant adverse reactions to food in infants and children. Partially and extensively hydrolyzed formulae are used for the therapy and prevention of cow's milk allergy. However, the immunogenic potency of hydrolyzed cow's milk formulae to induce and/or enhance the allergic phenotype in vivo is still under debate. The aim of this study was to assess the sensitizing capacity and residual allergenicity of various partially and extensively hydrolyzed cow's milk formulae in a murine model of cow's milk allergy. METHODS: BALB/c mice were immunized with either a cow's milk formula or various partially and extensively hydrolyzed formulae. Immediate cutaneous hypersensitivity reactions and allergen-specific antibody production were assessed. RESULTS: Although immunization with cow's milk resulted in 12/13 cases in a positive skin test response to intradermal injection of cow's milk formulae, only 1 mouse showed a positive skin test to one of the partially hydrolyzed formulae, and none showed positive reactions to other partially hydrolyzed formulae, any of the extensively hydrolyzed formulae, phosphate-buffered saline or the amino acid formula. However, 6 of 8 mice showed positive skin tests when immunized with partially hydrolyzed formulae and with one of the extensively hydrolyzed formulae. CONCLUSIONS: The residual allergenic potential is markedly reduced in many hydrolyzed formulae, but most of the formulae investigated could induce an allergic immune response in BALB/c mice. Our murine model seems to be suitable to investigate the sensitizing capacity of hydrolyzed formulae and to differentiate even between extensively hydrolyzed formulae.     

Comparison of intestinal anaphylactic reactions in sensitized mice challenged with untreated bovine milk and homogenized bovine milk

Outbred NMRI mice were sensitized for high IgE production either by subcutaneous injections of low doses of untreated bovine milk or homogenized bovine milk in combination with intraperitoneal injections of Freund's Complete Adjuvant or by oral administration of untreated or homogenized bovine milk without adjuvant. When analysed in murine passive cutaneous anaphylaxis test both types of milk resulted in production of reaginic antibodies against bovine milk proteins when given subcutaneously. When given orally, homogenized milk resulted in reagin production in 10 out of 14 mice, whereas untreated milk resulted in reagin production in only one out of 12 mice. The sensitized mice, and control mice, were orally challenged with either untreated milk, homogenized milk or 0.9% NaCl. Examination of the intestines 40 min after oral administration revealed that homogenized milk, contrary to untreated milk or 0.9% NaCl, resulted in a large increase in the mass of the proximal gut segment of mice sensitized orally with homogenized milk compared with control mice orally challenged with saline (P less than 0.001), and only mice both sensitized and challenged orally with homogenized milk showed degranulation of mast cells in the intestinal wall. By contrast, subcutaneously sensitized mice or mice sensitized orally with untreated bovine milk showed no significant intestinal reaction upon oral challenge with either homogenized or untreated bovine milk. These observations may indicate that the route of sensitization is of great importance when intestinal reactions are to be studied, and that homogenization of bovine milk may render the milk more aggressive with respect to its ability to induce intestinal reactions. The study indicates that mice may be an attractive experimental animal model for mimicking the intestinal anaphylactic reactions of cow milk-allergic humans.     

Effect of sensitization and anaphylactic shock on the plasma kinin system in guinea pigs

An increase in the circulating kininogen concentration was found in guinea pigs sensitized with human serum albumin or BCG vaccine. An anaphylactic reaction, subacute in character or manifested as shock with a marked disturbance of respiration, was accompanied by an increase of 1.5–2 times in kallikrein activity and of 6–10 times in plasma kininase activity. The mean decrease in the kininogen concentration was 20–48%, but when the shock followed an acute course it fell by 73–86%. The free kinin concentration was either raised or lowered.Department of Clinical Pathophysiology, M. F. Vladimirskii Moscow Regional Clinical Scientific-Research Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Fedorov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 78, No. 8, pp. 48–49, August, 1974.     

Evaluation of allergenicity of genetically modified soybean protein extract in a murine model of oral allergen-specific sensitization

Background With the development of genetically modified crop plants there has been a growing interest in the approaches available to assess the potential allergenicity of novel gene products. For additional assessment of the potential allergenicity of expressed proteins, informative data can be generated using animal models. Soybean is one of the major source of protein in human and animal nutrition, and has also been well characterized as a major allergenic source. Advances in biotechnology have resulted in an increasing number of genetically engineered foods, and among these soybean is one of the most widespread. Objective To develop and characterize a murine model of IgE‐mediated soybean sensitization induced by intragastric immunization, in the presence of Cholera Toxin, with wild‐type soybean extract (wt‐SE) or with genetically modified soybean extract (gm‐SE). Methods Balb/c mice born in our animal facilities, from females fed on soy‐free food, were fed with the same soy‐free food and used in all the experiments. Mice were sensitized by gavages with soybean extracts, and allergen‐specific IgE and IgG responses were studied by direct ELISA and ELISA inhibition. Antigen‐specific cell proliferation and cytokine production were evaluated in spleen cell cultures. Results Sensitization with both soybean extracts induced high levels of antigen‐specific IgE and IgG1 and low levels of specific IgG2a. Both wt‐SE and gm‐SE were able to inhibit the binding of specific IgE from mice immunized with gm‐SE to the same antigen used for the ELISA coating. A comparable proliferative response was obtained with the homologous as well as with the heterologous extracts. Conclusion In sensitized mice, we observed a predominantly T‐helper type 2 (Th2)‐type immune response, with increased soybean‐specific IgE and IgG1 antibodies and a concomitant increase of IL‐4 and IL‐5 production. Results obtained by specific IgE ELISA inhibition and by antigen‐specific T cell proliferation demonstrated that wt‐SE and gm‐SE shared B and T epitopes. The present murine model of soybean sensitization established by the oral route should provide valuable information about risk assessment for food allergy from new proteins of genetically modified foods.     

Effect of bilateral destruction of regions of the medial hypothalamus on the course of anaphylactic shock

Anaphylactic shock induced in rabbits after preliminary injury to various regions of the medial hypothalamus ran a more severe course than in the control. Regardless of the location of the foci of injury, in response to injection of the reacting dose of antigen the hypotensive response was more marked and the compensatory rise of blood pressure was slower. The severity of the anaphylactic shock depended on the time elapsing after injury to the hypothalamus until injection of the reacting dose of antigen.Allergological Research Laboratory, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. D. Ado.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 8, pp. 977–979, August, 1976.     

Effects of fermentation by Lactobacillus rhamnosus GG on the antigenicity and allergenicity of four cows' milk proteins

In this study, the effects of fermentation by Lactobacillus rhamnosus GG on the antigenicity and Immunoglobulin E (IgE)-binding inhibitions of α-lactalbumin (LA), β-lactoglobulin (LG), and α- caseins (CN) and β-CN were investigated, using polyclonal antibodies and milk-allergic patient sera in competitive enzyme-linked immunosorbent assay (ELISA). Meanwhile, the proteolysis of milk proteins was detected by trinitrobenzene sulfonic acid (TNBS) assay and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). As a result, we found that fermentation by L. rhamnosus GG could significantly reduce the antigenicity and IgE binding of α-LA, β-LG, α-CN and β-CN in reconstituted milk. In addition, the antigenicity of four proteins decreased to a lower value at 12 h of fermentation and at 0.5 d of cold storage after fermentation. And the IgE-binding inhibitions of four milk proteins in fermented milk were reduced and ranged from 6.5 to over 70% compared to raw milk.     

Enkephalins and anaphylactic shock: modulation and prevention of shock in the rat

Wistar rats were sensitized for anaphylactic shock with ovalbumin and treated intraperitoneally with 10 injections of methionine-enkephalin and leucine-enkephalin (4 mg/kg of body weight). Both pentapeptides suppressed the development of systemic anaphylaxis after intravenous injection of a shocking dose of ovalbumin. Methionine-enkephalin completely protected the animals from fatal shock. The level of circulating IgE and precipitating anti-ovalbumin antibodies decreased in rats given 10 injections of enkephalins. Mast cell degranulation was significantly inhibited in these animals as observed at autopsy. A single intraperitoneal injection of 4 mg/kg body weight of methionine-enkephalin given 30 min before the challenge with shock-inducing dose of antigen was also effective, although to a lesser extent, in protecting animals from anaphylactic shock. One injection of leucine-enkephalin did not exhibit significant antishock activity. The results suggest that enkephalins, and methionine-enkephalin in particular, are potent modulators of the complex biochemical processes underlying the anaphylactic shock in the rat.     

Determination of cow milk formula allergenicity in the rat model by in vitro mast cell triggering and in vivo IgE induction

The allergenicity of cow milk formulas was assessed in vitro by their ability to trigger prelabelled 3H-serotonin release from normal rat mast cells sensitized passively with rat reaginic anti-cow milk protein antisera. It was found that enzymatically hydrolyzed milk formulas had 1,000 (BEBAHA) to over 100,000 (ALFARE) times lower mast cell-triggering capacities than a standard milk formula (NAN). In vivo, these hydrolyzed formulas induced 100 (BEBAHA) to 10,000 (ALFARE) times less IgE antibodies than the standard milk formula.     

The effects of atropine on anaphylactic shock in the guinea-pig

The effects of atropine, 2 mg/kg i.v., on anaphylactic shock were studied in guinea-pigs sensitized to ovalbumin.Atropine only moderately reduced (–31%) the increase in pulmonary resistance observed and slightly prolonged (+26%) the survival time in pretreated animals compared with controls. These effects, however, were not statistically significant. The drug temporarily improved ventilation but had no influence on haematosis.On the other hand, atropine significantly reduced the amount of histamine released (–60%) and of GMPc synthetized in the lung (–21%). The levels of AMPc and prostaglandins E₁, E₂ and F₂ remained comparable to those of control animals.These results suggest that the reflex-induced action of the cholinergic system during anaphylaxis primarily affects large-calibre airways and that the role of acetylcholine in severe reactions is moderate when compared with the direct action of other mediators.     

Evaluation of the residual antigenicity and allergenicity of cow's milk substitutes by in vitro tests

BACKGROUND: This study aimed the to investigate presence of residual allergenic cow's milk proteins (CMP) in some milk substitutes employed in the treatment of cow's milk allergy (CMA). These allergens may interfere with the treatment, and elicit allergic reactions in sensitized individuals. METHODS: The protein composition of the different extracts was evaluated by Lowry's method and tricine SDS-PAGE. Different immunoenzymatic methods were used (ELISA, EAST and immunoblotting) to quantify total serum IgE and specific serum IgE, as well as to detect the presence of antigenic and allergenic components. RESULTS: The results showed a higher protein content in mammalian milks (cow, sheep, mare, goat, and human) than in hydrolyzed substitutes (partially or extensively hydrolyzed casein or whey proteins). Residual native, processed, or contaminant polypeptides have been identified in the moderate hydrolysates, whereas extensive hydrolysates did not show the presence of residual components by immunoblotting. However, specific antibodies with capacity to bind to peptides have been detected by EAST and ELISA, suggesting that extensive hydrolysates contain residual peptides that preserve immunoreactive epitopes. We were unable to demonstrate either residual antigenicity or allergenicity in an amino-acid-based formula. CONCLUSIONS: Immunoenzymatic methods were used to detect the presence of cross-reactive components in mammalian milks. Residual allergenic components from cow's milk could be identified in both the moderate and extensive hydrolysates analyzed. This information may be relevant to the treatment of CMA.