Depletion of mucosal mast cell protease by corticosteroids: effect on intestinal anaphylaxis in the rat.

Rats primed by infection with the intestinal nematode Nippostrongylus brasiliensis and challenged intravenously with soluble whole-worm antigen undergo systemic anaphylactic shock. The primary lesions are in the gut and include increased permeability of the mucosa together with release, into enteric secretions, of a mucosal mast cell (MMC)-specific serine proteinase, rat mast cell protease II (RMCP-II). This enzyme is also released into the blood of shocked rats. These manifestations of anaphylaxis were abolished in rats previously treated with corticosteroids (methylprednisolone acetate, 25 mg per kg of body weight, 48 and 24 hr before i.v. challenge with antigen). Suppression of the response was associated with depletion of RMCP-II and of MMC from the intestinal mucosa. Depletion occurred 4-24 hr after treatment with as little as 1 mg of methylprednisolone per kg. By contrast, neither connective tissue mast cells nor serum levels of parasite-specific IgE were depleted in rats given 2 X 25 mg of methylprednisolone per kg. The capacity of unprimed treated rats to mount passive cutaneous anaphylaxis was, however, impaired.     

Macrophage engulfment of mucosal mast cells in rats treated with dexamethasone

The effects of corticosteroid treatment on mucosal mast cells in rat jejunal mucosa were examined. Rats previously infected with Nippostrongylus brasiliensis received a single IP injection of 1 mg dexamethasone. Three hours later, one third of mucosal mast cells demonstrated minor granular changes (fusion or peripheral clear zones) by electron microscopy. At 7 hours, by light microscopy, the majority of mucosal mast cells appeared abnormal with clustering of granules. By electron microscopy, 151 of 233 (65%) mucosal mast cells had been engulfed by enlarged macrophages and were in various stages of degeneration inside large phagosomes. By 24 hours, the number of mucosal mast cells had decreased to less than 10% of the initial number with parallel decreases in tissue rat mast cell protease II and histamine levels. Serum levels of rat mast cell protease II did not increase, and intestinal morphology was invariably normal with no evidence of inflammatory changes up to and including 24 hours. Observations were similar in uninfected animals. In contrast, in rats undergoing antigen-induced anaphylaxis, a significant elevation of serum rat mast cell protease II level was evident at 3 and 7 hours, and macrophage engulfment of mucosal mast cells was never seen, although tissue edema, enterocyte loss, and hemorrhage were observed. It is concluded that dexamethasone treatment results in macrophage engulfment and destruction of mucosal mast cells that occurs without granular mediator release and local inflammatory effects.     

Long-term antigen challenge results in progressively diminished mucosal mast cell degranulation in rats

BACKGROUND & AIMS: The effects of short-term antigenic activation of mast cells on the gastrointestinal tract have been well characterized, but little is known about the effects of long-term exposure to antigen on mucosal mast cell reactivity. The aim of this study was to determine the effects of long-term antigen exposure on mucosal mast cell reactivity in the gastrointestinal mucosa. METHODS: Rats sensitized to chicken ovalbumin were orally challenged (short-term or long-term) with antigen. Rat mast cell protease II (RMCP-II) content was measured in serum as an index of mucosal mast cell degranulation. RESULTS: Short- term oral antigen challenge caused a 30-fold increase in serum RMCP-II levels. RMCP-II release was markedly diminished in long term-challenged rats (P < 0.001), despite increased tissue RMCP-II levels in stomach and jejunum. Although short-term antigen challenge significantly increased gastric acid secretion, no such response was observed after the long-term antigen challenge. In rats undergoing long-term challenge, a significant release of RMCP-II in response to intravenous antigen was not observed; however, mucosal mast cells remained responsive to intravenous anti-immunoglobulin E. CONCLUSIONS: Repeated activation of mucosal mast cells results in a progressive diminution of RMCP-II release not attributable to depletion of this mediator. This may represent an adaptive response aimed at minimizing the potentially deleterious effects of repeated exposure to an antigen. (Gastroenterology 1996 Dec;111(6):1516-23)     

Intestinal anaphylaxis in the rat. Effect of chronic antigen exposure

The effect of chronic dietary antigen challenge on the intestine was examined in sensitized rats. Three groups of Hooded-Lister rats were studied: animals sensitized to egg albumin; sham-sensitized animals; and unmanipulated controls. In sensitized rats, serum immunoglobulin E titers to egg albumin were greater than or equal to 1:64, whereas control and pair-fed rats showed no response. Sensitized rats received egg albumin 1 mg/ml in drinking water and rat chow ad libitum. Pair-fed animals also received egg albumin but were pair-fed with sensitized animals. Controls received water and rat chow ad libitum. Chronic antigen challenge resulted in reduced food intake and weight gain in sensitized animals. When the rats were killed after 9 days of antigen exposure, proximal intestine from experimental animals showed decreased disaccharidase activity, brush-border microvillus surface, area, and villus height. Crypt depth and enterocyte migration rate were increased. Mucosal mast cell involvement was suggested by mast cell proliferation, evidence of mast cell degranulation, and increased serum rat mast cell protease II levels. At the time of death, only sensitized jejunum demonstrated an increase in short-circuit current in Ussing chambers in response to antigen challenge. The findings indicate that chronic antigen exposure leads to intestinal injury, reduced food intake, and diminished weight gain.     

In vitro modelling of rat mucosal mast cell function in Trichinella spiralis infection

Intestinal infection with the parasitic nematode, Trichinella spiralis, provides a robust context for the study of mucosal mast cell function. In rats, mucosal mast cells are exposed to parasites during the earliest stage of infection, affording an opportunity for mast cells to contribute to an innate response to infection. During secondary infection, degranulation of rat mucosal mast cells coincides with expulsion of challenge larvae from the intestine. The goal of this study was to evaluate the rat bone marrow‐derived mast cells (BMMC) and the rat basophilic leukaemia cell line (RBL‐2H3) as models for mucosal mast cells, using parasite glycoproteins and antibody reagents that have been tested extensively in rats in vivo. We found that BMMC displayed a more robust mucosal phenotype. Although T. spiralis glycoproteins bound to mast cell surfaces in the absence of antibodies, they did not stimulate degranulation, nor did they inhibit degranulation triggered by immune complexes. Parasite glycoproteins complexed with specific monoclonal IgGs provoked release of rat mast cell protease II (RMCPII) and β‐hexosaminidase from both cell types in a manner that replicated results observed previously in passively immunized rats. Our results document that RBL‐2H3 cells and BMMC model rat mucosal mast cells in the contexts of innate and adaptive responses to T. spiralis.     

Ion transport abnormalities in inflamed rat jejunum. Involvement of mast cells and nerves

Basal and stimulated changes in ion transport in vitro were examined in jejunal mucosa from rats during inflammation produced after infection with the nematode Nippostrongylus brasiliensis. The gut was acutely inflamed at days 7 and 10 when net secretion of Na+ and Cl- ions was evident. Serum levels of rat mast cell protease II were elevated, providing evidence for mast cell activation. In addition, the magnitude of the short-circuit current responses to electrical transmural stimulation of enteric nerves (but not to histamine in the presence of neural blockade) were significantly reduced (p less than 0.01) to 17%-33% of control values, suggesting abnormalities of mucosal nerves. Following worm expulsion, serum levels of rat mast cell protease II and ion transport returned to normal. However, mastocytosis was apparent in gut mucosa and parasite antigen stimulated net secretion. In the absence of antigen, short-circuit current responses to nerve stimulation were increased (to 122% of controls; p less than 0.05). These findings suggest that changes in mast cells and enteric nerves occur during inflammation in this model and implicate neural and mast cell interactions with the epithelium in producing the ion-transport abnormalities.     

Colonic and jejunal motor disturbances after colonic antigen challenge of sensitized rat

BACKGROUND & AIMS: Inflammation in the colon may alter motility in the proximal gut and potentiate clinical symptoms. The aim of this study was to characterize the effect of colonic anaphylaxis on local (colonic) and remote (small intestinal) motility and identify the mechanism and mediators involved. METHODS: Rats were sensitized by intraperitoneal injection of 10 microg egg albumin and surgically prepared with electrodes in jejunum and colon and a colostomy tube. Colonic and jejunal myoelectric activity were recorded in fasted animals before and after colonic antigen challenge without and then after pretreatment with specific antagonists. RESULTS: Colonic antigen challenge of sensitized rats was associated with significant (1) increase in colonic myoelectric spike activity, (2) disruption of fasting jejunal motility and initiation of aborally propagating spike complexes, and (3) increase in plasma rat mast cell protease II levels with a decrease in granulated mast cells in colon but not jejunum. The myoelectric disturbance in both colon and jejunum was inhibited significantly by pretreatment with atropine and hexamethonium, doxantrazole, cyclooxygenase, and lipoxygenase inhibitors. Methysergide inhibited only the jejunal disturbance. CONCLUSIONS: Colonic antigen challenge of sensitized animals results in local mast cell activation and the release of mediators that modulate neural pathways to initiate both a local colonic and a remote jejunal myoelectric disturbance. (Gastroenterology 1997 Jun;112(6):1996-2005)     

Stem cell factor dependent hyperplasia of mucosal-type mast cells but not eosinophils in Schistosoma mansoni-infected rats

Stem cell factor (SCF) is a growth factor with multiple activities which acts on numerous cell types including primordial germ cells, haemopoietic stem cells, melanocytes and mast cells. SCF is critical for the development of the mast cell hyperplasia associated with infection with the intestinal parasites Nippostrongylus brasiliensis and Trichinella spiralis. In the present study we have assessed the role of SCF in the mast cell and eosinophil responses to Schistosoma mansoni in the rat by blocking its effects in vivo with polyclonal antibody to SCF. Rats treated with sheep anti-SCF antibody on days 21, 24, 27 and 30 of infection with S. mansoni showed a rapid decrease in serum concentrations of the mucosal mast cell-associated protease rat mast cell protease II (RMCP II) by day 24, compared with normal sheep IgG-treated controls. Similarly, the number of mucosal mast cells and RMCP II levels in both small intestine and liver were also significantly reduced by day 32 of infection. In contrast with the depeletion of mast cells and mast cell proteases, eosinophil numbers in liver or intestine did not change significantly after anti-SCF treatment compared with controls. These results confirm that mast cell survival and hyperplasia are dependent on the presence of SCF whilst demonstrating that the eosinophil recruitment to liver and intestine associated with S. mansoni infection is SCF-independent.     

Mediation of anaphylaxis-induced jejunal circular smooth muscle contraction in rats

Altered intestinal motility and diarrhea are features of food protein-induced intestinal anaphylaxis in the conscious rat. These experiments were performed to determine the mediator(s) responsible for jejunal circular smooth muscle contraction during this response. Hooded-Lister rats were sensitized by intraperitoneal injection of 10-g egg albumin, and controls were sham-sensitized with saline. Fourteen days later the contractility of the circular muscle in jejunal segments (mucosa intact) was examined in standard tissue baths in response to antigen (Ag) or other agents. While control and sensitized tissues contracted in similar fashion in response to stretch, bethanechol, histamine, or 5-hydroxytryptamine (5HT), Ag contracted only the segments of sensitized animals. The contractile response was: (1) specific to the sensitizing Ag, as bovine serum albumin did not induced contraction and (2) could be passively transferred with serum containing specific immunoglobulin E antibody (IgE-Ab). Concanavalin A, which degranulates both mucosal and connective tissue-type mast cells, and compound 48/80, which degranulates only connective tissue-type mast cells produced contractile responses. Ag-induced contraction was significantly inhibited by the mucosal and connective tissue-type mast cell stabilizer doxantrazole, but not the connective tissue mast cell stabilizer disodium cromoglycate. Diphenhydramine and cimetidine together significantly inhibited histamine-induced contraction, but failed to effect the Ag-induced contraction in sensitized tissues. While the contractile response to 5HT was reduced in the presence of methysergide (5HT₁-receptor antagonist), cinanserin (5HT₂-receptor antagonist), and ICS 205-930 (5HT₃-receptor antagonist), only cinanserin significantly inhibited the contractile response to Ag. Indomethacin significantly inhibited Ag-induced contraction. Ag-induced contraction was resistant to atropine and tetrodotoxin. Thus, food protein-induced alterations in intestinal motility in sensitized rats are due in part to an IgE-mediated mast cell degranulation, 5HT release, prostaglandin synthesis, and contraction of the circular smooth muscle.     

Relationship between mast cell degranulation and jejunal myoelectric alterations in intestinal anaphylaxis in rats.

The effects of two degranulators of mast cells and intestinal anaphylaxis on jejunal myoelectric activity were compared in rats fasted for 15 hours. Attempts to antagonize the motility changes were performed using antagonists of histamine and serotonin and a cyclooxygenase and lipoxygenase inhibitor. Hooded Lister rats were chronically fitted with electrodes implanted in the jejunal wall. A group of rats was sensitized to egg albumin and challenged 14 days later by intraduodenal infusion of antigen. Sensitized animals had serum titers greater than or equal to 1:64. The other group was administered with mast cells degranulators. Both 48/80 (1 mg/kg), a degranulator of connective mast cells, and bromolasalocid (2 mg/kg), acting on connective and mucosal mast cells, induced a phase of total spiking inhibition followed by a progressive irregular spiking activity until the recovery of migrating myoelectric complex pattern (about 3 hours after injection). In contrast, antigen challenge disrupted the migrating myoelectric complex pattern, which was replaced by a peculiar pattern characterized by propagated spike burst, lasting 98 +/- 11.3 minutes. Chlorpheniramine (1 mg/kg) antagonized only the inhibitory phase induced by degranulators and was ineffective on the intestinal anaphylaxis-induced motor changes. Methysergide (1 mg/kg) and indomethacin (5 mg/kg) significantly reduced the degranulator effects as well as the anaphylaxis-induced alterations of intestinal motility. It is concluded that anaphylaxis-induced motor disturbances are relevant to mucosal mast cell degranulation involving 5-hydroxytryptamine and arachidonic acid derivative products, whereas histamine release appears to be a minor component.     

Anaphylaxis inhibitory factor in IgE-dependent mast cell stimulation.

We have previously demonstrated that there is a factor present in some human serum which inhibits the passive cutaneous anaphylaxis reaction mediated by IgE. The present study analyzes the effect of this factor on mast cell IgE-dependent tumor necrosis factor (TNF)-alpha release. Rat peritoneal mast cells and RBL-2H3 cells treated with monoclonal mouse IgE anti-dinitrophenol (anti-DNP) followed by DNP-bovine serum albumin (DNP-BSA) were used and TNF-alpha release was measured at different time points. Similarly the percentage of rat peritoneal mast cell degranulation was determined. Results show a period of 30 min as optimal incubation time for TNF-alpha release in both mast cell populations. Human serum anaphylaxis inhibitory factor enriched fraction inhibited TNF-alpha release when it was in contact with IgE before the antigen treatment. Under these conditions the percentage of mast cell degranulation decreased. Mast cells incubated before IgE treatment with the factor alone do not release TNF-alpha and the percentage of degranulation increases due to a non-IgE-dependent process. A possible role of the inhibitory factor in the later phase reaction in addition to immediate hypersensitivity described previously is suggested.     

Basal secretion and anaphylactic release of rat mast cell protease-II (RMCP-II) from ex vivo perfused rat jejunum: translocation of RMCP-II into the gut lumen and its relation to mucosal histology.

The kinetics of the release of rat mast cell protease-II (RMCP-II) from mucosal mast cells in the jejunum of Nippostrongylus brasiliensis primed (immune) rats was investigated using ex vivo perfusion of a segment of jejunum through the cranial mesenteric artery. The aim of the study was to assess the role of the protease in anaphylaxis and in particular to ascertain whether it is responsible for the histological changes, which include widespread epithelial shedding, seen in the mucosa in in vivo models of anaphylaxis. Perfusion of the jejunal vasculature with a Krebs-Ringer buffer showed that there was basal secretion of RMCP-II by jejunal mast cells in all rats studied. The baseline concentration of RMCP-II was significantly greater (p < 0.05) in immune rats (> 7 ng/ml) previously exposed to nippostrongylus infection than in control, naive animals (< 2 ng/ml). Challenge of immune rats with 100 or 400 worm equivalents of whole worm antigen resulted in an immediate (within 40 seconds) and significant (p < 0.02) increase in the concentration of RMCP-II (to > 3 micrograms/ml) in the vascular perfusate, which was not seen in naive rats or immune rats challenged with an irrelevant antigen. Greater amounts of RMCP-II were also recovered from the jejunal lumen of immune rats compared with naive rats after challenge of both groups with worm antigen. Despite the release of microgram quantities of RMCP-II into the gut lumen and vascular perfusate, however, there were no significant changes seen in the mucosal histology. These results suggest that RMCP-II alone is not responsible fore the loss of gut epithelium seen during anaphylaxis in the rat.     

Colonoscopic allergen provocation (COLAP): a new diagnostic approach for gastrointestinal food allergy.

BACKGROUND: The clinical relevance of gastrointestinal food allergy in adults is largely unknown because the mechanisms are poorly understood and the diagnosis is difficult to confirm. AIMS: To improve the diagnostic means for confirming intestinal food allergy on an objective basis, a new colonoscopic allergen provocation (COLAP) test was developed. PATIENTS: The COLAP test was performed in 70 adult patients with abdominal symptoms suspected to be related to food allergy, and in five healthy volunteers. METHODS: During the COLAP test, the caecal mucosa was challenged endoscopically with three food antigen extracts, a buffer control, and a positive control (histamine). The mucosal weal and flare reaction was registered semiquantitatively 20 minutes after challenge, and tissue biopsy specimens were examined for mast cell and eosinophil activation. RESULTS: No severe systemic anaphylactic reactions were found in response to intestinal challenge. The COLAP test was positive to at least one food antigen in 54 of 70 patients (77%), whereas no reaction in response to antigen was found in healthy volunteers. Antigen induced weal and flare reactions were correlated with intestinal mast cell and eosinophil activation, as well as with patients' history of adverse reactions to food, but not with serum concentrations of total or specific IgE or skin test results. CONCLUSION: The COLAP test may be a useful diagnostic measure in patients with suspected intestinal food allergy and may provide a new tool for the study of underlying mechanisms.     

Regulation of intestinal mast cells and luminal protein release by cerebral thyrotropin-releasing hormone in rats

BACKGROUND & AIMS: Intestinal mast cell activity is modulated by the central nervous system, but the mechanisms are not well established. The aim of this study was to investigate whether cerebral thyrotropin- releasing hormone (TRH) activates intestinal mast cells and to elucidate the mechanisms involved, specifically, the contribution of mast cells to vagally stimulated luminal protein release. METHODS: In anesthetized rats, mast cell activation was assessed by measuring the release of the specific mucosal rat mast cell protease II (RMCP II) and prostaglandin (PG) D2 into the intestinal lumen. Luminal protein release was measured as an index of epithelial permeability to macromolecules. RESULTS: Intracisternal injection of the TRH analogue RX 77368 (30 ng) induced a transient increase in intestinal release of RMCP II and PGD2 that was abolished by dox-antrazole. RX 77368- stimulated RMCP II release was also abolished by vagotomy and reduced by atropine by 65%. However, both systemic capsaicin and indo-methacin enhanced RMCP II release. RX 77368-stimulated luminal protein release was abolished by vagotomy and reduced by doxantrazole. CONCLUSIONS: Central vagal activation by TRH stimulates intestinal mast cell secretion, in part via peripheral muscarinic receptors, and is modulated by PGs and capsaicin-sensitive afferent innervation. Intestinal mast cell activation contributes to the TRH analogue- stimulated luminal protein release. (Gastroenterology 1996 Dec;111(6):1465-73)     

Intestinal mucosal injury is associated with mast cell activation and leukotriene generation duringNippostrongylus-induced inflammation in the rat

We examined mucosal injury in the jejunum of the rat during infection with the nematode parasite, Nippostrongylus brasiliensis (Nb). Injury was documented morphologically (increase in crypt length with or without villus atrophy) and biochemically (activities of digestive or proliferative enzymes) and related to mast cell activation and leukotriene generation. At day 4 crypt length and thymidine kinase activity were increased; no changes in villus parameters were recorded. No evidence of mast cell activation was found and leukotriene levels in the mucosa were normal. At day 7, the gut was acutely inflamed and edema was present at the tips of the villi. This progressed to enterocyte detachment, resulting in villus atrophy with decreased activities of brush border enzymes. At this stage mucosal histamine was decreased and rat mast cell protease II (RMCP II) was increased in serum, indicating mast cell activation. In addition, mucosal leukotrienes (LTB4, LTC4, LTE4) were present in significant quantities. Following worm expulsion, the villus abnormalities resolved and serum RMCP II returned to normal. However, the crypt hyperplasia persisted. Our results suggest that during Nb infection at least two components of injury can be identified. One component, epithelial injury at the villus tips, may be related to activation of mucosal mast cells.     

Dosage de la tryptase : un guide d'utilisation pour le clinicien

Tryptase is the most abundant endopeptidase released by mast cells degranulation, involved in many pro and anti-inflammatory processes. Normal serum tryptase range is 0–11.4 μg/L. Tryptase is a useful diagnostic tool for anaphylaxis, systemic mastocytosis (SM) and mast cell activation syndrome (MCAS), where specific threshold values must be used. SM diagnosis criteria include evidence of dense mast cell infiltrate either in the bone marrow or the affected organ (such as skin), presence of KIT D816V mutation and elevated serum tryptase level (>20 μg/L). In SM, tryptase level is correlated with the burden of mast cells in bone marrow. MCAS should be considered in case of severe and recurrent typical clinical signs of systemic mast cell activation involving at least two organs, associated with an increase in serum tryptase level of 20% + 2 μg/L from the individual's baseline. Anaphylaxis is the most severe among hypersensitivity reactions. A clonal mast cell disorder is a central question in anaphylaxis and appropriate explorations should be conducted in these patients. Triggers for anaphylactic reactions vary significantly in the general population and in patients with MS or MCAS. Finally, physicians must be aware of the many pathological and physiological situations that affect tryptase levels.     

Stem cell factor contributes to intestinal mucosal mast cell hyperplasia in rats infected with Nippostrongylus brasiliensis or Trichinella spiralis, but anti-stem cell factor treatment decreases parasite egg production during N brasiliensis infection

We assessed the effects of the c-kit ligand, stem cell factor (SCF), in the jejunal mucosal mast cell hyperplasia that occurs during infection with the intestinal nematodes, Nippostrongylus brasiliensis or Trichinella spiralis in rats. Compared with vehicle-treated rats, rats treated with SCF (25 micrograms/kg/d, intravenous [i.v.] for 14 days) during N brasiliensis infection exhibited significantly higher levels of the rat mucosal mast cell (MMC)-associated protease, rat mast cell protease II (RMCP II) in the jejunum and serum on day 8 of infection, but not on days 10 or 15 of infection. By contrast, in comparison to rats treated with normal sheep IgG, rats treated with a polyclonal sheep antirat SCF antibody exhibited markedly decreased numbers of jejunal MMCs, levels of jejunal RMCP II, and serum concentrations of RMCP II during infection with either nematode, particularly at the earlier intervals of infection (< or = day 10). Taken together, these findings indicate that SCF importantly contributes to MMC hyperplasia and/or survival during N brasiliensis or T spiralis infection in rats, but that levels of endogenous SCF are adequate to sustain near maximal MMC hyperplasia during infection with these nematodes. Notably, treatment of rats with SCF somewhat increased, and treatment with anti- SCF significantly decreased, parasite egg production during N brasiliensis infection. This finding raises the interesting possibility that certain activities of intestinal MMCs may contribute to parasite fecundity during infection with this nematode.     

The immune response during a Strongyloides ratti infection of rats

A range of immune parameters was measured during a primary infection of Strongyloides ratti in its natural rat host. The immune parameters measured were interleukin-4 (IL-4) and interferon-gamma from both the spleen and mesenteric lymph node (MLN) cells; parasite-specific immunoglobulin G(1)(IgG(1)), IgG(2a) and IgG(2b) in serum and in intestinal tissue; parasite-specific IgG and total IgE in serum; parasite-specific and total IgA in intestinal tissue and rat mast cell protease II in intestinal tissue. Parasite-specific IgG(1), IgG(2a) and total IgE in serum and parasite-specific IgA and rat mast cell protease II in intestinal tissue all occurred at significantly greater concentrations in infected animals, compared with non-infected animals. Similarly, the production of IL-4 by MLN cells stimulated with parasitic female antigen or concanavalin A occurred at significantly greater concentrations in infected animals, compared with non-infected animals. In all, this suggests that there is a T-helper 2-type immune response during a primary S. ratti infection. These data also show the temporal changes in these components of the host immune response during a primary S. ratti infection.     

Resident mast cells are the main initiators of anaphylactic leukotriene production in the liver.

During anaphylaxis the sensitized liver can have substantial capacity for leukotriene production. However, the intrahepatic cellular source for these potent eicosanoid mediators has been unclear so far. We therefore analyzed the appropriate role of resident liver cells in organ-specific generation of leukotrienes by immunohistochemical localization of 5-lipoxygenase, by measurement of cysteinyl leukotriene production in animals or isolated livers and by histochemical monitoring of mast cells in rat, guinea pig and mouse livers, respectively. During anaphylaxis in vivo, these species all generated large amounts of leukotrienes. Immunohistochemistry with rat liver demonstrated resident mast cells as the predominant cell type in liver containing 5-lipoxygenase. Rat and guinea pig livers contained numerous mast cells and produced substantial amounts of leukotrienes on antigen challenge; in contrast, mouse livers neither showed detectable mast cells nor generated leukotrienes when stimulated analogously. Infusion of histamine or serotonin (1 mmol/L each) or of the degranulating substance P (8 mumol/L) did not elicit leukotriene generation in rat livers. Furthermore, substantial degranulation of liver mast cells by compound 48/80 (0.5 mg/kg body mass) was paralleled by only modest leukotriene formation (63 +/- 10 pmol in bile/kg body mass/30 min). These results indicate that during anaphylaxis mast cells are the main intrahepatic cells initiating leukotriene production and that such leukotriene generation is likely to be independent of mast cell degranulation or the release of histamine or serotonin.     

Nonatopic allergen-independent mast cell activation in parasitized eosinophilic athymic rats

It is possible to induce eosinophilia in congenitally athymic rats by infection with the parasite Ascaris suum. Athymic bronchial eosinophilia is associated with increased expression of IL-5 and eotaxin mRNA, and with the presence of residual T cells and mast cells. Anamnestic mastocytosis is particularly pronounced and in this study we examine the relationship between mast cell degranulation and IgE production in athymic rats following infection. Incubation of peritoneal mast cells from athymic rats with anti-IgE induced dose-dependent degranulation, as measured by histamine release. However, the failure of mast cells from infected athymic rats to degranulate following incubation with all but one of the parasite antigens selected confirms the absence of a specific IgE response. In contrast, all agonists induced degranulation in euthymic rats. The only parasite-derived factor to induce histamine release in all mast cells was Ascaris body fluid, which contains an element capable of inducing IgE-independent degranulation. Furthermore, increased levels of rat mast cell protease II in athymic bronchoalveolar lavage fluid suggest degranulation of mast cells by IgE and allergen independent means in vivo. We believe that the development of eosinophilic lung inflammation in the absence of specific IgE makes this a prototype for investigating the immunological mechanisms underlying nonatopic asthma.