Local Anaphylaxis Research Articles

Nutritional Value and Antigenicity of Two Milk Protein Hydrolysates in Rats and Guinea Pigs

Two milk protein (whey protein and casein) hydrolysates were obtained by enzymatic hydrolysis. In the casein hydrolysate, an ultrafiltration stage was performed to remove all the peptides with molecular weights > 2500. A nutritional value study of both hydrolysates was undertaken. No differences were found between the native proteins and their enzymatic hydrolysates. Neither the enzymatic hydrolysis nor the heat treatments used affected the nutritional value of the protein sources. The two hydrolysates were studied for antigenic properties. The enzymatic hydrolysis of the whey protein concentrate followed by a thermal treatment at 90 degrees C for 10 min reduced significantly its antigenicity. However, the in vivo allergenicity tests showed some positive reactions in both systemic anaphylaxis and passive cutaneous anaphylaxis. The casein hydrolysate, after being ultrafiltered, was devoid of sensitizing capacity by the oral route and was also ineffective in producing local or systemic anaphylaxis in previously sensitized animals, as shown by in vivo assays.

Medical consequences of multiple fire ant stings occurring indoors

Stings by the imported fire ant almost always lead to dermal wheal and flare reactions followed by sterile pustules at sting sites. Less commonly, large local dermal reactions, pyoderma, anaphylaxis, or neuropathy may occur. Such reactions have previously been associated with contact with the insects out of doors. We present two previously unreported cases of indoor attacks on individuals by imported fire ants. One patient experienced a cerebrovascular accident in association with the attack, whereas the second patient had no obvious sequelae. With these two reports, a total of four such indoor massive sting episodes have appeared in the recent medical literature. Physicians and other individuals living in areas indigenous to the fire ant should be aware that infestation of buildings with fire ants may be associated with attacks on human beings indoors. Individuals with cognitive dysfunction seem to be especially at risk for attacks by fire ants.

Intestinal nematode infections in children: the pathophysiological price paid

The mechanism by which small animals such as rodents resist or eliminate nematode parasites requires mucosal inflammation as the final effector of the immune response. The resulting freedom from chronic infection may be worth the price of short-term illness. Putative vaccines which attempt to enhance the natural effect will have to take into account the inflammatory cost to the host. Human helminthiases involve a more stable equilibrium between host and parasite. The medical literature on hookworm disease and clinical ascariasis describes, for the former, some chronic inflammatory effects correlated with worm burden, but for the latter a less quantified or predictable set of detrimental effects. We describe a current, systematic study of the inflammatory response to whipworm infection, in which anaemia, growth retardation and intestinal leakiness are viewed as predictable consequences related to infection intensity. There is evidence for the absence of cell-mediated immunopathology. However, a specific, IgE-mediated local anaphylaxis may, at least partly, mediate the deleterious effects. Increased numbers of mucosal macrophages may also contribute to the chronic, systemic effects through their output of cytokines. Similar attempts to show the mechanisms of pathogenesis and quantify the effects of hookworm disease should be undertaken.

Immunoregulation of Endometrial and Jejunal Epithelia Sensitized by Infection

The hypothesis was tested that the uterus of the rat orally infected with the parasite Trichinella spiralis becomes hypersensitized and that subsequent antigenic challenge affects functions in the endometrial epithelium. Results of experiments comparing the immunological responsiveness of isolated rat uterus with that of the jejunum supports our hypothesis. Antigenic challenge of uterus mounted in Ussing-type chambers causes an elevation in transuterine short circuit current (Isc) of 6.4 +/- 0.8 microA/cm2. The transduction of the antigenic signal to elicit the electrophysiological response involves 5-hydroxytryptamine (5-HT) working through a nerve-independent pathway. The antigen-stimulated rise in Isc peaks approximately 3 min after challenge. The uterine response is blocked by diisothiocyanostilbene-2,2'-disulfonic acid, an inhibitor of bicarbonate-chloride exchange. The antigen-evoked change in jejunal Isc is biphasic, peaking at 1.5 and approximately 4.0 min after challenge, and is about 10-fold greater in magnitude than the Isc in the uterus. The transductive pathway in the jejunum involves 5-HT, histamine and prostaglandin acting partly through intrinsic nerves. The jejunal response to antigen is inhibited by diphenylamine-2-carboxylate, a chloride channel blocker. Changes in net ion transport which are primed by infection and evoked by antigen are apparently triggered by local anaphylaxis in both the uterus and jejunum.

Involvement of Type I Hypersensitivity in Rapid Rejection of Trichinella spiralis from Adult Rats

The role of type I hypersensitivity in the rapid rejection of Trichinella spiralis from actively and passively immunized rats was examined. Net intestinal fluid secretion, which occurs during the rejection of the parasites, was used to verify the expression of local anaphylaxis and was examined for its possible role in the rejection process. Worm establishment in the small intestine 30 min after intraduodenal inoculation was significantly reduced in rats that were passively immunized with immune serum containing anti-Trichinella IgE as compared with recipients of normal serum. Worm-induced fluid secretion in immune rats was completely inhibited by the combined action of indomethacin and diphenhydramine. However, worm rejection was not affected. L-651,392, a 5-lipoxygenase inhibitor, also failed to prevent rejection. Ketanserin (a serotonin S2 receptor antagonist) and MDL-72222 (a serotonin S3 receptor antagonist) together blunted the rapid rejection response and reduced fluid secretion. Furthermore, intra-arterial perfusion of serotonin into nonimmune rats caused fluid secretion and reduced worm establishment. In nonimmune rats prostaglandin E2, cholera toxin, and hypertonic Krebs-mannitol solution were used to evoke the same level of intestinal fluid secretion as that induced by reinfection in immune rats or by serotonin in nonimmune hosts. When larvae were inoculated into the secreting gut, their infectivity was unaffected. The results suggest that anaphylaxis is involved in the rapid rejection of T. spiralis in immune rats and that serotonin is a possible chemical mediator of worm rejection. Although the mode of action of serotonin in the rejection process remains unknown, its possible involvement through fluid secretion can be ruled out.

Immunological control of drug absorption from the gastrointestinal tract: the mechanism whereby intestinal anaphylaxis interferes with the intestinal absorption of bromthymol blue in the rat.

Rats were immunized intraperitoneally with ovalbumin and the disappearance of bromthymol blue (BTB) from the intestinal lumen, its accumulation in the tissue, and its net absorption were examined by means of an in-situ recirculation technique during local anaphylaxis. The disappearance of BTB from the intestinal lumen and its net absorption were significantly reduced, but there was no significant effect on its accumulation in the tissue. The pH value of the luminal solution and the perfusate volume were not influenced by intraluminal challenge with the antigen in ovalbumin-immunized rats. In addition, no significant effect was observed on intestinal permeability to BTB in the in-vitro everted sac technique. The intestinal blood flow, measured by a hydrogen clearance method, was not reduced significantly by the intraluminal exposure to antigen. There was enhanced Evans Blue leakage and mucus release in the perfusate after intraluminal challenge with ovalbumin in ovalbumin-immunized rats, but not in non-immunized rats. A significant increase of BTB binding with macromolecular substances in the perfusate was observed during the local anaphylaxis. These findings suggest that the decreased absorption of BTB is due to the interaction with the macromolecular substances in the perfusate during local anaphylaxis.

Anamnestic stimulus-specific myoelectric responses associated with intestinal immunity in the rat

Changes in small intestinal motility reflective of local anaphylaxis are hypothesized to be associated with the rapid, immune-mediated rejection of infective larvae of the nematode Trichinella spiralis. This hypothesis was tested by comparing intestinal myoelectric activity in primarily and secondarily infected rats; the latter group represented immunized hosts, i.e., immunized by initial contact with the parasite. Patterns of slow waves, action potentials, interdigestive migrating myoelectric complexes, and migrating action-potential complexes induced by secondary infection differed from those associated with primary infection. Patterns in both cases differed from those in uninfected rats. Secondary but not primary infection evoked changes in myoelectric patterns within 15 min after administration of infective larvae. These changes could not be elicited by inoculation with dead larvae, larval excretory-secretory antigens, or a heterologous parasite, Eimeria nieschulzi. Results indicate that altered smooth muscle contractile activity is part of an anamnestic, stimulus-specific response to T. spiralis in immunized rats. These changes may represent the in vivo equivalent of the anaphylactically mediated intestinal smooth muscle contraction elicited in vitro in the Schultz-Dale reaction.

Allergenicity of Whey Protein

Enzymatic in vitro hydrolysis was evaluated as a possible treatment to abolish the allergenicity of whey proteins in view of their use in infant formulas. Guinea pigs without prior immunological contact (including fetal life) with cow's milk were fed various preparations of cow's milk proteins. Oral exposure to milk or untreated whey protein led to anaphylactic sensitization of the animals. In contrast, trypsin-hydrolyzed whey protein and a peptide preparation produced from the tryptic hydrolysate by ultrafiltration were devoid of sensitizing capacity by the oral route. The hydrolysate (crude or purified) was also ineffective in triggering local or systemic anaphylaxis in previously sensitized animals.

Allergenicity of Whey Protein

Enzymatic in vitro hydrolysis was evaluated as a possible treatment to abolish the allergenicity of whey proteins in view of their use in infant formulas. Guinea pigs without prior immunological contact (including fetal life) with cow'S milk were fed various preparations of cow'S milk proteins. Oral exposure to milk or untreated whey protein led to anaphylactic sensitization of the animals. In contrast, trypsin‐hydrolyzed whey protein and a peptide preparation produced from the tryptic hydrolysate by ultrafiltration were devoid of sensitizing capacity by the oral route. The hydrolysate (crude or purified) was also ineffective in triggering local or systemic anaphylaxis in previously sensitized animals.

Immunological control of drug absorption from the gastrointestinal tract: Effect of local anaphylaxis on the intestinal absorption of low molecular weight drugs in the rat.

Intestinal absorption of various drugs was examined by means of in situ recirculation technique during local anaphylaxis. The antibody was determined by passive cutaneous anaphylaxis technique in rats immunized once or three times. The optimal condition of local anaphylaxis was determined by the leakage of Evans Blue. The most significant increase in leaks of the dye was observed by the intraluminal challenge with 400 mg of ovalbumin for 10 min in ovalbumin-immunized rats, and this condition was chosen as the optimal condition of local anaphylaxis. Under this condition, intestinal absorption of caffeine, phenylbutazone, and bromthymol blue (BTB) significantly decreased in ovalbumin-immunized rats compared with the control, whereas no significant effect was noted in the intestinal absorption of salicylic acid, quinine, pralidoxime iodide (2-PAM), tetracycline, and phenol red. In normal rats, no significant decrease was obtained in the intestinal absorption of caffeine, phenylbutazone, and BTB. On the other hand, the decreased absorption of BTB was not found in ovalbumin-immunized rats by the intraluminal challenge with bovine gamma-globulin. Furthermore, there was no significant change in the decreased absorption of BTB between rats immunized once and three times. The most effective condition for decreased BTB absorption was observed by the intraluminal challenge with 200 mg of ovalbumin for 10 min in ovalbumin-immunized rats, which almost correlated with the data of Evans Blue leakage. From these observations, it appears that the mucosal immune responses affect the intestinal absorption of low molecular weight drugs.

Histamine release in acute anaphylactic enteropathy of the rat.

Investigations into the role of allergic enteropathy in acute and chronic intestinal inflammation have been hampered by the lack of objective confirmation for intestinal mast cell activation. Utilizing an established model of acute allergic enteropathy in the rat, we report the enhanced intraluminal recovery of the mast cell mediator histamine after in vivo antigen challenge in sensitized animals. The enhanced histamine recovery is dose dependent, antigen-specific, and restricted to that segment of bowel challenged, thus confirming local intestinal anaphylaxis. The progression of histologic enteropathy is documented and shown to correlate with the entry of mast cells into the intestinal lumen during, but not before, the anaphylactic response. Pretreatment of the sensitized animal with prostaglandin E2 or doxantrazole, but not cromolyn, significantly inhibits the anaphylactic response.

Local ocular anaphylaxis to papain enzyme contained in a contact lens cleansing solution

This is a case report of a patient who developed IgE-mediated sensitization and subsequently ocular angioedema and conjunctivitis from papain contained in a commercial contact lens cleansing solution. Serum-specific IgE and positive cutaneous prick tests to papain and chymopapain were detected. When the lens solution containing papain was stopped, there was resolution of her allergic symptoms. Recognition of this route of papain-induced sensitization may be important in those patients undergoing chemonucleolysis with chymopapain who may be at greater risk to develop a systemic allergic reaction after injection of this enzyme.

Macrophages in ocular tissues of rats. Determination of their number after local anaphylaxis and other procedures.

The number of macrophages in rat ocular tissues was determined, with the tip of the eyelid containing about 3,000 macrophages per cubic millimeter, which is similar to the number of mast cells at this site. Fewer macrophages were present in orbital tissues and conjunctiva. Macrophages accumulated in ocular tissues of immunized rats injected locally with antigen, but the number did not exceed that observed in antigen-injected controls. Injection of various fluids into ocular tissues, but not the trauma of needle punctures alone, stimulated a marked accumulation of macrophages. Thus, the response to nonspecific stimuli masked the macrophage response to antigen-induced, ocular anaphylaxis.

Investigation of leukotriene involvement in the vasopermeability response associated with guinea pig tracheal anaphylaxis: Comparison with cutaneous anaphylaxis

A direct comparison of the role of leukotrienes in mediating the increase in microvascular permeability associated with guinea pig tracheal and cutaneous anaphylaxis was obtained by simultaneous administration of inflammatory stimuli to both trachea and ear. The SRS-A antagonist, FPL 55712, reduced the increase in tracheal extravascular permeability response associated with local anaphylaxis. Moreover, the inhibitory effect of the histamine H 1-receptor antagonist, mepyramine, was not augmented by the additional presence of FPL 55712. In contrast to tracheal anaphylaxis, a distinct leukotriene component was indicated in cutaneous anaphylaxis since the mepyramine-FPL 55712 combination produced a greater inhibition than mepyramine alone. These results suggest that the degree of leukotriene involvement in anaphylaxis may vary between tissues.

A new method of testing anti-allergic drugs.

Rats were sensitized with a single intraperitoneal dose of bovine serum albumin in alhydrogel plus Bordetella pertussis vaccine, and local anaphylaxis was elicited in the paw by soluble antigen 2 weeks later. The response was mainly due to IgE-type antibodies and proved to be highly sensitive to beta-adrenoceptor agonists. Dexamethasone inhibited the response after a lag phase. Methysergide, disodium chromoglycate, diethylcarbamazine, BW 775/c, nordihydroguarjaretic acid, and FPL 55712 were also suppressive, while mepyramine was without effect. A synergism between methysergide and FPL 55712 was shown. Active local paw anaphylaxis appears to be adequate and easily applicable for large-scale screening of anti-allergic drugs.

Effect of locally induced intestinal anaphylaxis on the uptake of a bystander antigen

Adult Sprague-Dawley rats weighing 250 gm were immunized with 100 μg egg albumin (EA) and alum. Fourteen days later, intestinal anaphylaxis was induced by intraduodenal or intragastric challenge with EA. To test for the changes in vascular and mucosal permeability that accompany anaphylaxis, the rats were injected intravenously with 125 I-rat serum albumin (RSA) at the time of challenge and the amount of radioactivity retained in the wall of gut segments, as well as the trichloroacetic acid (TCA)-precipitable radioactivity present in the secretions obtained from these segments, was determined. Enhanced retention of 125 I-RSA was found in the wall of the duodenum and first and second halves of the small intestine, and increased amounts of TCA-precipitable radioactivity was found in gut segments obtained from antigen-challenged compared with buffer-challenged rats. In other EA-immunized rats, bovine serum albumin (BSA) was administered by gavage 1 hr before challenge with EA. Increased amounts of immunoreactive BSA were detected by radioimmunoassay in the serum of rats subjected to local intestinal anaphylaxis. Possible consequences of the enhanced systemic uptake of protein present during local intestinal anaphylaxis were considered. It was suggested that such proteins might induce an IgE antibody response, thereby broadening the anaphylactic sensitivity of the animal to additional antigens.

936 GUT INDUCED ANAPHYLAXIS AND UPTAKE OF A BYSTANDER PROTEIN: AN AMPLIFICATION OF ANAPHYLACTIC SENSITIVITY

This study sought to test the effect of intestinal anaphylaxis on uptake of an unrelated antigen. Adult rats were immunized with 100 ug egg albumin (EA) and alum. Fourteen days later, intestinal anaphylaxis was induced by intraduodenal or intragastric challenge with EA. To test for the changes in vascular and mucosal permeability with anaphylaxis, rats were given 125I-RSA intravenously on challenge and the radioactivity retained in the wall of the gut segments and the trichloracetic acid (TCA)-precipitable radioactivity present in secretions from these segments was determined. Enhanced retention of 125I-RSA was found in the wall of the gut segments and increased amounts of TCA-precipitable radioactivity was found in secretions from antigen-challenge compared to control rats. In other EA immunized rats, bovine serum albumin (BSA) was administered by gavage one hour before challenge with EA. Increased amounts of immunoreactive BSA was detected by radioimmunoassay in serum of rats subjected to local intestinal anaphylaxis. Among the consequences of enhanced systemic uptake of protein present during intestinal anaphylaxis is the induction of an IgE antibody response to this protein. Preliminary experiments suggest that an IgE antibody response can be induced by this mechanism. These findings suggest the broadening of anaphylactic sensitivity of animals to new antigens and may explain multiple sensitivities to food antigens in humans.

Expulsion of Nippostrongylus brasiliensis by mice deficient in mast cells.

Expulsion of the intestinal helminth, Nippostrongylus brasiliensis, occurs spontaneously about 2 weeks after a primary infection of rats and mice. Cellular changes in the small intestine coincident with the period of expulsion have suggested several mechanisms by which this 'self-cure' may be effected. Local anaphylaxis was proposed as a possible means of parasite clearance; this hypothesis has been supported by the demonstration of specific reaginic antibody production and jejunal mast cell accumulation in infected animals. In addition, increased mucus secretion and more recently, goblet cell proliferation in the jejunal mucosa of rats have been noted and considered as potentially important in mediating the self-cure reaction. The data presented below indicate that in the absence of demonstrable mast cells, the course of a primary infection with this parasite is unchanged; however, they are supportive of a role for globlet cells in the self-cure reaction.

Reversed response to histamine following local anaphylaxis in blood-perfused calf trachea in situ

Tracheas of calves were perfused via the tracheal artery in situ. An endotrcheal tube recorded tracheal muscle contractions. Carbachol, histamine or antigen caused tracheal vascular depressor responses. Carbachol caused contraction; histamine, 2-methylhistamine and antigen relaxed the tracheal muscle. Following antigen challenge, vascular responses remained unchanged; however endotracheal relaxations to histamine were reversed (i.e. contractions). Allergenic ‘challenge’ impairs H 2-receptors (airway relaxation), thus diminishing a compensatory mechanism which may relate to hyperresponsiveness characteristic of asthma.

Quantitation of immediate and delayed hypersensitivity responses in Trichinella-infected mice. Correlation with worm expulsion.

The developmental relationships between active cutaneous or local anaphylaxis, delayed hypersensitivity and worm expulsion were quantitatively examined in Trichinella-infected mice. The onset of both types of hypersensitivities and increase in sensitivity to antigen following an initial infection correlated with the onset and rate of elimination of adult worms. Mice passively sensitized with serum containing both IgG1 and IgE antibodies, but not IgG1 alone, expelled their worms at a faster rate in comparison to the controls. On the basis of these findings it is suggested that local allergic reactions mediated by anaphylactic antibodies are involved in the development of resistance to Trichinella infection.