Anaphylactic Reactions In Guinea Pigs Research Articles

Preclinical safety evaluation of low molecular weight heparin-deoxycholate conjugates as an oral anticoagulant.

The preclinical safety of a newly developed oral anticoagulant, the low molecular weight heparin-deoxycholate conjugate (OH09208), was evaluated by a comprehensive evaluating program in compliance with standard guidelines. The single dose oral toxicity study in rats receiving 2000 and 5000 mg kg(-1) of OH09208 did not reveal any mortality, unusual body weight changes or necropsy findings. The results of the 4-week oral toxicity study with a 4-week recovery program in rats receiving OH09208 in doses of 100, 300 and 1000 mg kg(-1) day(-1) did not reveal any mortality, or indicate any unusual clinical signs, or show any toxicokinetic relationships to the administration of OH09208. Although the increase in liver enzymes in one male dog treated with 300 mg kg(-1) day(-1) and one female dog treated with 1000 mg kg(-1) day(-1) could not be excluded from the effect of the test substance, no other toxicologically significant changes were observed in the 4-week oral toxicity study with a 4-week recovery in beagle dogs. Thus, while the no-observed-adverse-effect level value from the 4-week study in both male and female rats was 1000 mg kg(-1) day(-1), those from the 4-week study in male and female beagle dogs were 300 and 1000 mg kg(-1) day(-1), respectively. Furthermore, OH09208 did not induce anaphylactic reactions in guinea pigs, micronucleated bone marrow cells in male ICR mice, chromosomal aberration in Chinese hamster lung cell lines, bacterial reverse mutation, and any abnormalities in hERG current assay, mouse central nervous system and dog cardiovascular studies. Overall, there were no unexpected toxicities in this preclinical study that might have precluded the safe administration of OH09208 to humans.

Antiallergenic activity of birch bark dry extract with at least 70% betulin content

Birch bark dry extract (BBDE) containing at least 70% betulin and betulin alone were compared with the reference drugs Suprastin and Claritin for their antiallergenic action. It is established that BBDE and betulin reduce by half the systemic anaphylactic reaction in guinea pigs immunized with ovalbumin from chicken egg white, their action being comparable with that of Suprastin in mild and medium anaphylactic shock. The course of BBDE administration per os or i.p. leads to a significant decrease in the content of IgE antibodies to ovalbumin in mice of various strains immunized in different regimens. The effect of BBDE on anaphylactic contraction of isolated guinea pig ileum samples was evaluated and showed no direct action upon the H1-receptors. Studies on models of pseudoallergenic reaction to concanavalin A in mice and carragenan-induced paw edema in rats showed that the anti-inflammatory action of BBDE is comparable with that of Suprastin and Claritin and is more pronounced than the effect of betulin.

Role of neurogenic inflammation in antigen-induced vascular extravasation in guinea pig trachea.

Conflicting results have been reported about the role of sensory nerves in the allergen-induced plasma extravasation in sensitized guinea pigs using capsaicin desensitization. To investigate the role of tachykinins released from sensory nerves in the anaphylactic reaction in guinea pigs in vivo, we used a selective inhibitor of neutral endopeptidase, phosphoramidon, and a selective neurokinin (NK)-1 receptor antagonist, CP-96,345. Male Hartley guinea pigs were sensitized to OVA by two i.p. injections (70 mg) at 1-wk intervals. Two wk later, the animals were anesthetized and OVA was administered for 2 min by aerosol through a tracheal cannula. Plasma extravasation was assessed by the photometric measurement of the extravasated Evans blue after formamide extraction. Administration of aerosolized OVA to sensitized guinea pigs increased dye extravasation in the trachea in a dose-dependent manner, an effect that was demonstrable 5 min after exposure to allergen and that reached a maximum 10 min after exposure. At 5 min after OVA (5%), phosphoramidon (2.5 mg/kg, i.v.) did not increase the amount of dye in the trachea significantly and CP-96,345 (4 mg/kg) did not decrease the extravasated dye. At 10 min after OVA, allergen-induced plasma extravasation was potentiated by phosphoramidon by 56%, and was inhibited by CP-96,345 (4 mg/kg) by 42%. In the presence of phosphoramidon, CP-96,345 reduced the OVA-evoked plasma extravasation at 10 min in a dose-related manner (0.1-4 mg/kg). CP-96,345 (4 mg/kg) did not affect plasma extravasation induced by platelet-activating factor (100 nmol/kg, i.v.). These results suggest that tachykinin release from sensory nerves after allergen challenge to airways in sensitized guinea pigs is not responsible for the early increase in plasma extravasation, but, tachykinin release appears to play an important role in the subsequent extravasation.

The study of host defense mechanisms of the nose. Third report: absorption of an antigen from the nasal mucosa

Nasal provocation test, by placing the allergen disc on the nasal mucosa of the patient with pollinosis consecutively, brought easily the systemic symptoms such as cough, periorbital edema, and urticaria. Similarly, the sensitized guinea pigs showed a change upon respiratory examination after the nasal administration of a large doses of the allergen. On the other band, an extract of Bordetella bronchiseptica induced an anaphylactic reaction in guinea pigs and γ1 antibody participated in this reaction. After the nasal administration of radioisotope labeled extract of the bacteria, radioactivity in the blood of guinea pigs sensitized with the extract, was lower than that of non-sensitized guinea pigs. These facts suggest that the nasal mucosa could absorb a high molecular substance such as an antigen, and that the existence of the antibody in nasal mucosa has a great influence on absorption of the antigen from nasal mucosa.

Antigenicity of semisynthetic penicillin preparations to evoke systemic anaphylactic reactions in animal models.

Four commercially available penicillin preparations (benzylpenicillin, aminobenzylpenicillin, carbenicillin and sulbenicillin) and two kinds of the new semisynthetic penicillin preparations (mezlocillin and piperacillin) evoked systemic anaphylactic reactions in guinea pigs immunized with one of the following penicillin-Ascaris extract (As) conjugates in aluminum hydroxide gel (alum), i.e., benzylpenicilloyl (BPO)-As, aminobenzylpenicilloyl (ABPO)-As, carbenicilloyl (CBPO)-As, sulbenicilloyl (SBPO)-As, mezlocillin-As conjugate (MEZ-As) and piperacillin-As conjugate (PIP-As). The tracheal chains prepared from the tracheas of the guinea pigs immunized with BPO-As, MEZ-As, or PIP-As revealed anaphylactic contractions in vitro on addition of benzylpenicillin, mezlocillin, or piperacillin preparations. These results indicated that the semisynthetic penicillin preparations as well as benzylpenicillin preparation have enough antigenicity to evoke systemic anaphylactic reactions in guinea pigs immunized with penicillin-As conjugate. Such guinea pigs seem to be suitable animals for investigations of penicillin allergy.

Suppression of Anaphylactic Reactions in Guinea Pigs by Previous Treatment with Ragweed Aerosol

Guinea pigs subjected to intranasal dusting with ragweed (RW) pollen or intranasal instillation of RW extract (RWE) showed anaphylactic reactions when subjected to RW extract aerosol (RWE). These animals, however, did not respond to an aerosol of RW whole pollen. Animals which received the RWEA treatment prior to nasal dusting with RW pollen showed no subsequent anaphylactic reactions when exposed to RWEA. Both the aerosol-sensitive and the aerosol-nonresponsive animals exhibited high titers of homocytotropic antibodies to RWE. In addition, the aerosol-nonresponsive animals also had precipitating antibodies of IgG type. The state of nonresponsiveness did not appear to be due to the precipitating antibodies, since this condition could not be passively transferred by sera and blocking antibody activity was not demonstrable.

In vitro release of eosinophil chemotactic factor of anaphylaxis (ECF-A) from guinea pig skin.

An in vitro release of the eosinophil chemotactic factor of anaphylaxis (ECF-A) in anaphylactic reactions was studied using skin slices from actively sensitized guinea pigs. ECF-A and histamine were released in vitro from actively sensitized guinea pig skin by antigen challenge. The magnitude of ECF-A release did not always parallel that of anaphylactic histamine release in guinea pig skin, suggesting that there may be no correlation between the amount of ECF-A and that of histamine released from the skin in anaphylactic reactions in guinea pigs.

Benzylpenicillin preparations can evoke a systemic anaphylactic reaction in guinea pigs

All of the five commercially available benzylpenicillin preparations obtained from different sources and a PcG preparation prepared by filtration of a commercial PcG on Sephadex G10 elicited the systemic anaphylactic reactions in guinea pigs which had been immunized with benzylpenicilloyl (BPO)- Ascaris extract conjugate (BPO-As) mixed with aluminum hydroxide gel. These preparations could evoke no such reactions in guinea pigs immunized with BPO-bovine gamma globulin conjugate (BPO-BGG) emulsified with complete Freund's adjuvant. The severity of the systemic anaphylactic reactions correlated significantly with the titers of either 8-day passive cutaneous anaphylactic (8-day PCA) reactions or 4-hr PCA reactions evoked with the same benzylpenicillin preparations. In vitro benzylpenicillin preparation contracted the tracheas of the guinea pigs immunized with BPO-As. These results indicated that the commercially available benzylpenicillin preparations have enough antigenicity to evoke systemic anaphylactic reactions in guinea pigs immunized with BPO-As mixed with aluminum hydroxide gel. Such guinea pigs represent an animal model for investigation of penicillin allergy.

Inhibition of anaphylactic reaction in guinea pigs by antibodies to fragments of Fc from autologous IgG.

The effect of antibodies to Fc subfragments of guinea pig IgG on the anaphylactic reaction in guinea pigs has been studied. Control animals were immunized with guinea pig IgG and Fab, human IgG and albumin. All animals with antibodies to fragments of autologous Fc survived challenge with horse serum, whereas the control animals died in anaphylactic shock.

Evidence for the Domain Hypothesis: Location of the Site of Cytophilic Activity toward Guinea Pig Macrophages in the CH3 Homology Region of Human Immunoglobulin G

Abstract Most of the effector functions of human immunoglobulin G (IgG), such as complement fixation, transplacental transfer, cytophilic activity toward macrophages and control of catabolic rate are mediated through the Fc region of the molecule. Some of these functions have been shown more recently to be associated with particular subfragments of the Fc region corresponding to either the CH2 or CH3 homology regions of the IgG1 heavy chain. Minta and Painter (1) showed that the pFc' fragment which comprises the CH3 homology region was able to interfere with the passive cutaneous anaphylactic reaction in guinea pigs, presumably by competing with antibody for sites on the cutaneous mast cells. Kehoe and Fougereau (2) showed that fragments from the CH2 region of a mouse immunoglobulin were able to activate the complement system when suitably aggregated. More recently Ellerson et al. (3) showed that a fragment comprising the whole CH2 homology region of human IgG was able to activate the complement system when aggregated on polystyrene latex.

Depletion of Plasma Complement in Vivo by a Protein of Cobra Venom: Its Effect on Various Immunologic Reactions

Abstract The purified low molecular weight anticomplementary factor (CoF) in the venom of the cobra Naja naja was tested for its behavior in vivo and for its effect on various components of complement in several species of animals and the development of several immunologic reactions. CoF complexed with a plasma protein immediately after injection, the sedimentation rate of the complex being about 9S. The complex which inactivated purified C3 circulated with a half-life of 32 hr in rabbits. High blood levels of the CoF complex and minimal side effects were obtained using multiple injections of CoF over a 24-hr period. Under these conditions hemolytic complement levels fell to less than 10% of normal values in rabbits, rats and guinea pigs. C3 was markedly diminished in these species, and in guinea pigs C3 levels, as revealed in immune adherence, fell to less than 1% of normal. C3 and hemolytic complement levels remained low for 4 to 6 days. In two species tested, C5 protein was also lost during CoF treatment and in rabbits C6 activity diminished about half to one third of normal. The effect of CoF treatment in several immunologic diseases was assayed. Lesions depending upon neutrophil participation for full development were markedly inhibited. In acute nephrotoxic nephritis neutrophil accumulation in glomeruli and proteinuria were diminished. Arthus reactions, especially in the rat, were also inhibited. By contrast, immunologic reactions not requiring neutrophils, i.e., homologous cutaneous anaphylactic reactions in guinea pigs and contact (delayed) sensitivity were not affected by CoF treatment. Other possible effects of CoF in vivo were studied. The numbers of formed elements of the blood were not markedly altered. The most consistent finding was a modest elevation in neutrophils. Hemolysis was avoided by treatment of animals with multiple small doses of CoF. Neutrophils responded normally to immune adherence and chemotactic stimuli. Kininogen levels and clotting tests were unchanged.

A PENICILLOYLATED PROTEIN IMPURITY AS A SOURCE OF ALLERGY TO BENZYLPENICILLIN AND 6-AMINOPENICILLANIC ACID

Fractionation of the sodium salt of 6-aminopenicillanic acid (6-A.P.A.) led to the isolation of a high-molecular-weight material which gave strong passive cutaneous anaphylactic reactions in guineapigs. The " purified " 6-A.P.A. no longer elicited a response. It seems likely that this impurity derived from the Escherichia coli preparation used in the production process. A similar proteinaceous impurity was found in commercially available benzylpenicillin. A method for purifying benzylpenicillin was developed. On storage however, a high-molecular-weight substance developed and this may be derived from the penicillin molecule by polymerisation. Thus, not all the immunological activity of the penicillins is due to the penicilloylated protein. It is thought that the administration of purified benzylpenicillin will reduce the level of induction of hypersensitivity and may decrease the number of allergic manifestations if given inadvertently to a penicillin-sensitive patient. Purified benzylpenicillin should not, however, be deliberately given to a patient known to be sensitive. The implications for oral therapy are not yet known.

Comparative investigations of purified diphtheria and tetanus T-components and their fragments

1. By means of Rivanol ®, ammonium sulphate and zone electrophoresis it was possible to isolate highly purified T-components from tetanus- and diphtheria-horse hyperimmune sera which sediment in the ultracentrifuge homogeneously with S7. The 7 S T-components were digested with pepsin to homegeneous 5 S fragments from which by reduction with mercaptoethanol and fractionation on Sephadex 3.5 S fragments were obtained. 2. From the 7 S preparations and the fragments amino acid, carbohydrate analyses and fingerprints were made. They showed differences between the 7 S and 5 S preparations. The 3·5 S fragments differed from the 5 S fragments only by their content in cystine. 3.7 S, 5 S and 3·5 S preparations were effective as toxine neutralizing antibodies. The 3·5 S preparations, however, showed a lower specific activity. 4. The T-components and their fragments proved to be species specif1. They showed, however, quantitative differences. The homologous antigens provoked stronger anaphylactic reactions in guinea pigs. 5. In amino acid, fingerprints and carbohydrate analyses no differences have been found between diptheria and tetanus 7 S T-components and fragments. These findings, however, do not allow the conclusion, whether the antibody specificity is based on the primary structure or not. The amino acid analysis suggests that the T-component is more related to the γG-globulin than to the γA-globulin fraction of human serum.

II. Reverse Passive Cutaneous Anaphylaxis in the Guinea Pig with Ovalbumin

Summary Reverse passive cutaneous anaphylaxis with ovalbumin and rabbit anti-ovalbumin antibody has been regularly produced in guinea pigs. The quantity of antibody necessary to elicit such reactions was found to be the same as that required to sensitize guinea pig skin for direct anaphylaxis in a few minutes after the intravenous injection of antibody. It may be concluded that in explanation of anaphylactic reactions in guinea pigs in the direct as well as the reverse procedure, since antibody “fixation” appears to be unnecessary in sensitizing guinea pig tissues, one can invoke an antigen-antibody system giving rise to complexes toxic for the guinea pig, providing some quantitative conditions have been respected.

ANTIALLERGIC ACTIVITY AND SOME CHEMICAL PROPERTIES OF AN ANTIHISTAMINE PRINCIPLE IN HUMAN AND ANIMAL TISSUE EXTRACTS

In continuation of earlier studies it has been observed that extracts prepared from animal and human tissues such as gingiva, colon, brain, lung, breast, rectum, stomach, placenta, exert antihistamine activity in both in vitro and in vivo experiments; gingiva and lung showed the highest potency. It has also been observed that these extracts can protect against both the in vitro anaphylactic reaction (Schultz-Dale reaction) and the in vivo anaphylactic reaction in guinea pigs previously sensitized to egg white. This antihistamine activity appears to be due to a relatively simple chemical compound containing hydroxyl and carbonyl groups with characteristic chromatographic and spectral properties, although its exact structure has not been established.In view of the widespread distribution of this antihistamine factor, it is suggested that it might be of physiological significance as an antiallergic substance and warrants further study as a potential new drug for allergic diseases. These and other aspects of the problem are being explored.

ANTIALLERGIC ACTIVITY AND SOME CHEMICAL PROPERTIES OF AN ANTIHISTAMINE PRINCIPLE IN HUMAN AND ANIMAL TISSUE EXTRACTS

In continuation of earlier studies it has been observed that extracts prepared from animal and human tissues such as gingiva, colon, brain, lung, breast, rectum, stomach, placenta, exert antihistamine activity in both in vitro and in vivo experiments; gingiva and lung showed the highest potency. It has also been observed that these extracts can protect against both the in vitro anaphylactic reaction (Schultz-Dale reaction) and the in vivo anaphylactic reaction in guinea pigs previously sensitized to egg white. This antihistamine activity appears to be due to a relatively simple chemical compound containing hydroxyl and carbonyl groups with characteristic chromatographic and spectral properties, although its exact structure has not been established.In view of the widespread distribution of this antihistamine factor, it is suggested that it might be of physiological significance as an antiallergic substance and warrants further study as a potential new drug for allergic diseases. These and other aspects of the problem are being explored.

Molecular Bases of Passive Sensitization

Summary The effect of chemical procedures designed to break the disulfide linkages of γ-globulins on passively sensitize guinea pig skin was studied with a human γ-globulin-rabbit anti-HGG system. Human (antigen) and rabbit (antibody) γ-globulins lost sensitizing activity after treatment with mercaptoethanol followed by sulfhydryl blocking agents such as iodoacetate or iodoacetamide, as determined by passive cutaneous anaphylaxis tests in guinea pigs. Treatment with either mercaptoethanol or iodoacetate alone did not affect their sensitizing activity. Mercaptoethanol-iodoacetate treated proteins, either as antigen or antibody elicited cutaneous anaphylactic reactions in guinea pigs previously sensitized with unmodified γ-globulins as antigen or antibody and also showed precipitin reactions with the complementary reagents comparable to reactions using unmodified proteins. The complement-fixing properties of the HGG-anti-HGG systems were unaffected by the treatment of either antigen or antibody by mercaptoethanol and iodoacetate. The results suggested that irreversible cleavage of disulfide linkages in human and rabbit γ-globulin molecules was correlated with a decrease in their affinity for guinea pig tissues.

Tissue mast cells, histamine and 'slow reacting substance' in anaphylactic reaction in guinea pig.

AbstractAnaphylactic reaction was produced in vitro in isolated guinea‐pig tissue. Histamine and ‘slow reacting substance’ (‘SRS’) in different tissues occurred in amounts proportional to the mast‐cell population. The occurrence of these substances was attended with a reduction of the number of tissue mast cells. When the appearance of histamine and ‘SRS’ was blocked by enzyme inhibitors, there was no decrease in mast cells. It is suggested that the tissue mast cells may be the common source of both histamine and ‘SRS’ in anaphylactic reaction.

The mechanism of histamine release in anaphylactic reaction in guinea pig and rat.

AbstractChakravarty, N. The mchanism of histamine release in anaphylactic reaction in guinea pig and rat.The mechanism of histamine release was studied in anaphylactic reaction in vitro in tissues from guinea pig and rat; it was compared, in rat, with histamine release induced by compound 48/80. The observations suggest that histamine release in both species is mediated by the activation of an enzyme system probably with essential sulphydryl and amino groups. Histamine release in anaphylaxis and that induced by compound 48/80 in rat appear to be mediated by the same mechanism. Some differences were observed in anaphylactic histamine release in the two species: anoxia inhibited histamine release in guinea‐pig tissue but not in rat tissue; the same was true of two triphenylmethane dyes. Despite these differences, anaphylactic histamine release in the two species seems to follow, on the whole, a common pathway.

Immunochemistry of Synthetic Polypeptides and Polypeptidyl Proteins

Summary One polyglutamic acid, a polylysine, and a multichain polypeptide caused the formation of antibodies in rabbits which would not precipitate the homologous polypeptide but did form precipitates with bovine albumin, fibrinogen, and a few other naturally occurring proteins. The reactions between the modified proteins and antibodies appeared to be especially sensitive to salt concentration—precipitation being more easily inhibited by high salt concentration than the precipitins of many natural proteins. With a few other polyglutamic acid preparations, specific precipitates were formed with their antisera when the ionic strength was lowered to 0.05, suggesting that physiologic saline may have prevented the precipitin formation. The rabbit antisera to polyglutamic acid and the multichain polypeptide caused only a weak anaphylactic reaction in guinea pigs when the challenging antigen was the homologous or similar polypeptide and caused a moderate to fatal anaphylactic shock in guinea pigs challenged with bovine albumin or polyglutamyl bovine albumin. The synthetic polypeptides did not inhibit the reactions between their antisera and the proteins when tested by inhibition of precipitation techniques and passive anaphylaxis. No evidence of a strong reaction between the polypeptides and antisera to similarly modified proteins was found in inhibition of precipitation methods and passive anaphylaxis. Incomplete antibodies were not detected in antisera to polyglutamic acids which did not contain precipitating antibodies. A complex between the multichain polypeptide and its antiserum was not detected in electrophoresis and ultracentrifugal experiments.