Evidence that the endothelin A receptor can enhance IgE-dependent anaphylaxis in mice
2011; Elsevier BV; Volume: 128; Issue: 2 Linguagem: Inglês
10.1016/j.jaci.2011.04.012
ISSN1097-6825
AutoresMartin Metz, Beatrix Schäfer, Mindy Tsai, Marcus Maurer, Stephen J. Galli,
Tópico(s)Drug-Induced Adverse Reactions
ResumoTo the Editor:Anaphylaxis is a life-threatening, systemic allergic reaction that can occur suddenly after contact with an otherwise innocuous substance. In certain subjects previously sensitized to a particular antigen, re-exposure to even very small amounts of that antigen can induce the rapid and extensive activation of mast cells (MCs) and basophils. The ensuing massive release of MC- and basophil-derived mediators can then result in severe and sometimes fatal pathophysiological responses.1Summers C.W. Pumphrey R.S. Woods C.N. McDowell G. Pemberton P.W. Arkwright P.D. Factors predicting anaphylaxis to peanuts and tree nuts in patients referred to a specialist center.J Allergy Clin Immunol. 2008; 121 (e2): 632-638Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar It is generally accepted that anaphylaxis in human subjects is mediated by activation of MCs and basophils induced by the cross-linking of FcεRI, the high-affinity receptor for IgE, for example, with IgE and specific bivalent or multivalent antigen. Although IgE can also bind to the IgG receptors FcγRII and FcγRIII and to galectin-3, which are expressed on some MC populations, IgE is thought to influence MC and basophil functions in patients with anaphylaxis mainly through its interaction with FcεRI.1Summers C.W. Pumphrey R.S. Woods C.N. McDowell G. Pemberton P.W. Arkwright P.D. Factors predicting anaphylaxis to peanuts and tree nuts in patients referred to a specialist center.J Allergy Clin Immunol. 2008; 121 (e2): 632-638Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar However, it is likely that factors in addition to antigen-specific IgE can contribute to the occurrence or severity of anaphylaxis. For example, many allergic patients exhibit high levels of circulating antigen-specific IgE but never have anaphylaxis, whereas other subjects with low concentrations of specific IgE in the blood can experience anaphylactic shock.1Summers C.W. Pumphrey R.S. Woods C.N. McDowell G. Pemberton P.W. Arkwright P.D. Factors predicting anaphylaxis to peanuts and tree nuts in patients referred to a specialist center.J Allergy Clin Immunol. 2008; 121 (e2): 632-638Abstract Full Text Full Text PDF PubMed Scopus (147) Google ScholarEndothelin-1 (ET-1), a 21-amino-acid naturally occurring peptide with potent vasoconstrictor activity, has been reported to exacerbate certain allergic reactions in rodents. For example, increased levels of ET-1 have been described in animal models of active anaphylaxis,2Sanchez-Cifuentes M.V. Rubio M.L. Ortega M. Largo R. Gomez-Garre M.D. Gonzalez Mangado N. et al.Endothelin-1 expression during early response after antigen challenge in brown-Norway rats.Pulm Pharmacol Ther. 1998; 11: 215-219Crossref PubMed Scopus (5) Google Scholar, 3Shigematsu T. Miura S. Hirokawa M. Hokari R. Higuchi H. Tsuzuki Y. et al.Endothelins promote egg albumin-induced intestinal anaphylaxis in rats.Gastroenterology. 1998; 115: 348-356Abstract Full Text Full Text PDF PubMed Scopus (22) Google Scholar and pharmacological inhibition of the endothelin receptor ETA reportedly attenuated the magnitude of ovalbumin–induced intestinal anaphylaxis3Shigematsu T. Miura S. Hirokawa M. Hokari R. Higuchi H. Tsuzuki Y. et al.Endothelins promote egg albumin-induced intestinal anaphylaxis in rats.Gastroenterology. 1998; 115: 348-356Abstract Full Text Full Text PDF PubMed Scopus (22) Google Scholar or paw edema4Sampaio A.L. Rae G.A. D’Orleans-Juste P. Henriques M.G. ETA receptor antagonists inhibit allergic inflammation in the mouse.J Cardiovasc Pharmacol. 1995; 26: S416-S418PubMed Google Scholar in ovalbumin–sensitized animals. ET-1 can induce degranulation of MCs in mice,5Maurer M. Wedemeyer J. Metz M. Piliponsky A.M. Weller K. Chatterjea D. et al.Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (256) Google Scholar, 6Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google Scholar suggesting one mechanism by which increased levels of ET-1 might exacerbate anaphylaxis. However, the potential interactions between MC and ET-1 in anaphylaxis are complex. For example, mouse MCs can secrete ET-16Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google Scholar; MC-derived proteases, particularly MC-derived carboxypeptidase A,5Maurer M. Wedemeyer J. Metz M. Piliponsky A.M. Weller K. Chatterjea D. et al.Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (256) Google Scholar, 7Metz M. Piliponsky A.M. Chen C.C. Lammel V. Ǻbrink M. Pejler G. et al.Mast cells can enhance resistance to snake and honeybee venoms.Science. 2006; 313: 526-530Crossref PubMed Scopus (285) Google Scholar, 8Schneider L.A. Schlenner S.M. Feyerabend T.B. Wunderlin M. Rodewald H.R. Molecular mechanism of mast cell mediated innate defense against endothelin and snake venom sarafotoxin.J Exp Med. 2007; 204: 2629-2639Crossref PubMed Scopus (124) Google Scholar can hydrolyze ET-1 to a biologically inactive peptide by removing the C-terminal tryptophan8Schneider L.A. Schlenner S.M. Feyerabend T.B. Wunderlin M. Rodewald H.R. Molecular mechanism of mast cell mediated innate defense against endothelin and snake venom sarafotoxin.J Exp Med. 2007; 204: 2629-2639Crossref PubMed Scopus (124) Google Scholar; and pretreatment of in vitro–derived mouse MCs with ET-1 can significantly decrease the cells’ degranulation in response to IgE and antigen in vitro.6Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google Scholar In the present study we investigated whether the severity of IgE-dependent passive systemic or cutaneous anaphylaxis is altered in mice treated with BQ-123, an antagonist of the endothelin receptor ETA.MCs are thought to represent a critical effector cell in IgE-dependent anaphylaxis.1Summers C.W. Pumphrey R.S. Woods C.N. McDowell G. Pemberton P.W. Arkwright P.D. Factors predicting anaphylaxis to peanuts and tree nuts in patients referred to a specialist center.J Allergy Clin Immunol. 2008; 121 (e2): 632-638Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar Prior functional assays and expression analyses of various MC populations demonstrated that ETA can be expressed by connective tissue–type MCs, including peritoneal MCs.5Maurer M. Wedemeyer J. Metz M. Piliponsky A.M. Weller K. Chatterjea D. et al.Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (256) Google Scholar, 6Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google Scholar, 9Metz M. Lammel V. Gibbs B.F. Maurer M. Inflammatory murine skin responses to UV-B light are partially dependent on endothelin-1 and mast cells.Am J Pathol. 2006; 169: 815-822Abstract Full Text Full Text PDF PubMed Scopus (46) Google Scholar However, immature bone marrow–derived cultured mouse mast cells (BMCMCs), which have some similarities to mucosal-type MCs, are largely unresponsive to ET-1 and have very low levels of receptor expression.6Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google ScholarWe confirmed, using flow cytometric analysis, the substantial expression of ETA and weak expression of ETB on C57BL/6 mouse peritoneal MCs (Fig 1, A), as well as the absence of detectable surface ETB and very weak expression of ETA on C57BL/6 mouse BMCMCs (Fig 1, B). When BMCMCs were induced to undergo further maturation by maintaining the cells in the presence of IL-4 and stem cell factor, in addition to IL-3, the expression of ET receptors increased to levels approaching those of freshly isolated peritoneal MCs (Fig 1, C). Taken together, these results support the notion that a robust surface expression of ETA is a characteristic of mature murine MCs.Binding of ET-1 to ETA can induce strong and rapid degranulation of MCs,5Maurer M. Wedemeyer J. Metz M. Piliponsky A.M. Weller K. Chatterjea D. et al.Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (256) Google Scholar, 6Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google Scholar and ET-1 levels previously have been shown to be increased in certain animal models of anaphylaxis.2Sanchez-Cifuentes M.V. Rubio M.L. Ortega M. Largo R. Gomez-Garre M.D. Gonzalez Mangado N. et al.Endothelin-1 expression during early response after antigen challenge in brown-Norway rats.Pulm Pharmacol Ther. 1998; 11: 215-219Crossref PubMed Scopus (5) Google Scholar, 3Shigematsu T. Miura S. Hirokawa M. Hokari R. Higuchi H. Tsuzuki Y. et al.Endothelins promote egg albumin-induced intestinal anaphylaxis in rats.Gastroenterology. 1998; 115: 348-356Abstract Full Text Full Text PDF PubMed Scopus (22) Google Scholar We therefore hypothesized that ET-1 can have effects on MC-mediated allergic reactions. To investigate whether ET-1 can contribute to features of severe IgE- and MC-dependent allergic reactions, we first analyzed IgE-dependent passive systemic anaphylaxis in mice treated with the selective ETA antagonist BQ-123. Mice were sensitized intraperitoneally with IgE anti–2,4-dinitrophenol (DNP; in 100 μL of sterile, pyrogen-free 0.9% NaCl [saline]) and challenged intraperitoneally 24 hours later with 1 mg of DNP–human serum albumin (HSA; 30-40 DNP:1 HSA; Sigma, St Louis, Mo) in 100 μL of saline to induce anaphylaxis. Mice were pretreated with BQ-123 (Calbiochem, San Diego, Calif; 20 nmol in 200 μL of vehicle [saline + 0.5% dimethyl sulfoxide]) or 200 μL of vehicle intraperitoneally 45 minutes before challenge. Mice treated with BQ-123 exhibited a statistically significant, although modest, amelioration of the allergic reaction, as assessed by monitoring the ensuing change in body temperature (Fig 2, A). Furthermore, the reduced hypothermia was associated with a modest but statistically significant reduction in the extent of degranulation of peritoneal MCs, as assessed morphologically (Fig 2, B),5Maurer M. Wedemeyer J. Metz M. Piliponsky A.M. Weller K. Chatterjea D. et al.Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (256) Google Scholar, 7Metz M. Piliponsky A.M. Chen C.C. Lammel V. Ǻbrink M. Pejler G. et al.Mast cells can enhance resistance to snake and honeybee venoms.Science. 2006; 313: 526-530Crossref PubMed Scopus (285) Google Scholar indicating that blockade of ETA results in decreased activation of MCs in this setting. In support of this conclusion, pretreatment with BQ-123 in mice subjected to passive systemic anaphylaxis also resulted in reduced levels of both MC-derived mouse MC protease 1 in the serum (Fig 2, C) and histamine in the peritoneal lavage fluid (Fig 2, D).Fig 2Selective blockade of ETA attenuates IgE-dependent anaphylactic reactions. A-D, Body temperature (Fig 2, A), peritoneal MC degranulation (Fig 2, B), levels of mouse mast cell protease 1 (mMCP-1) in serum (Fig 2, C), and levels of histamine in peritoneal lavage fluid (PLF; Fig 2, D) in C57BL/6 mice sensitized with IgE anti-DNP intravenously and then injected 24 hours later with DNP-HSA or vehicle (Veh) intraperitoneally 45 minutes after intraperitoneal injection of either BQ-123 or vehicle. E and F, Ear swelling (Fig 2, E) and extravasation of Evans blue dye (Fig 2, F) 1 hour after DNP-HSA challenge in mice injected intradermally with IgE anti-DNP and challenged intraperitoneally 24 hours later with DNP-HSA (in saline plus 1% Evans blue dye). Mice were injected intraperitoneally with either BQ-123 or vehicle 10 minutes before DNP-HSA challenge. Fig 2, A, ∗∗∗P < .005 versus the corresponding IgE anti-DNP plus DNP plus vehicle-challenged group (ANOVA). Fig 2, B, ∗P < .05 and ∗∗∗P < .005 for the indicated comparisons (χ2 test). Fig 2, C-F, ∗P < .05 versus the corresponding vehicle-treated group (unpaired, 2-tailed Student t test). All results are presented as means ± SEMs (Fig 2, A) or means + SEMs (Fig 3, B-F), with 8 to 10 mice per group in each panel (see the expanded Fig 2 legend in this article’s Online Repository at www.jacionline.org for more information about methods).View Large Image Figure ViewerDownload Hi-res image Download (PPT)To assess the possible contribution of ETA to IgE- and MC-dependent allergic responses in the skin, we next induced passive cutaneous anaphylaxis by means of intradermal injection of IgE anti-DNP (20 μg in 100 μL of saline) and then challenged the mice intraperitoneally with DNP-HSA (1 mg in 100 μL of saline) or 100 μL of saline alone 24 hours later. The increase in ear thickness and the extent of plasma extravasation were then quantified as measures of allergic inflammation. Injection of BQ-123 intravenously before elicitation of the allergic reaction again resulted in a significant reduction in the assessed parameters, with about 30% to 40% reduction in the measured responses compared with those seen in the identically sensitized and challenged but vehicle-treated mice (Fig 2, E and F).Taken together, our findings show that treatment with the ETA antagonist BQ-123 can ameliorate systemic or local IgE- and anaphylactic responses in mice. We hypothesize that one mechanism that might contribute to this observation is that the ETA antagonist can reduce ETA-dependent enhancement of MC activation by endogenous ET-1 present during the allergic reaction. To the Editor: Anaphylaxis is a life-threatening, systemic allergic reaction that can occur suddenly after contact with an otherwise innocuous substance. In certain subjects previously sensitized to a particular antigen, re-exposure to even very small amounts of that antigen can induce the rapid and extensive activation of mast cells (MCs) and basophils. The ensuing massive release of MC- and basophil-derived mediators can then result in severe and sometimes fatal pathophysiological responses.1Summers C.W. Pumphrey R.S. Woods C.N. McDowell G. Pemberton P.W. Arkwright P.D. Factors predicting anaphylaxis to peanuts and tree nuts in patients referred to a specialist center.J Allergy Clin Immunol. 2008; 121 (e2): 632-638Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar It is generally accepted that anaphylaxis in human subjects is mediated by activation of MCs and basophils induced by the cross-linking of FcεRI, the high-affinity receptor for IgE, for example, with IgE and specific bivalent or multivalent antigen. Although IgE can also bind to the IgG receptors FcγRII and FcγRIII and to galectin-3, which are expressed on some MC populations, IgE is thought to influence MC and basophil functions in patients with anaphylaxis mainly through its interaction with FcεRI.1Summers C.W. Pumphrey R.S. Woods C.N. McDowell G. Pemberton P.W. Arkwright P.D. Factors predicting anaphylaxis to peanuts and tree nuts in patients referred to a specialist center.J Allergy Clin Immunol. 2008; 121 (e2): 632-638Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar However, it is likely that factors in addition to antigen-specific IgE can contribute to the occurrence or severity of anaphylaxis. For example, many allergic patients exhibit high levels of circulating antigen-specific IgE but never have anaphylaxis, whereas other subjects with low concentrations of specific IgE in the blood can experience anaphylactic shock.1Summers C.W. Pumphrey R.S. Woods C.N. McDowell G. Pemberton P.W. Arkwright P.D. Factors predicting anaphylaxis to peanuts and tree nuts in patients referred to a specialist center.J Allergy Clin Immunol. 2008; 121 (e2): 632-638Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar Endothelin-1 (ET-1), a 21-amino-acid naturally occurring peptide with potent vasoconstrictor activity, has been reported to exacerbate certain allergic reactions in rodents. For example, increased levels of ET-1 have been described in animal models of active anaphylaxis,2Sanchez-Cifuentes M.V. Rubio M.L. Ortega M. Largo R. Gomez-Garre M.D. Gonzalez Mangado N. et al.Endothelin-1 expression during early response after antigen challenge in brown-Norway rats.Pulm Pharmacol Ther. 1998; 11: 215-219Crossref PubMed Scopus (5) Google Scholar, 3Shigematsu T. Miura S. Hirokawa M. Hokari R. Higuchi H. Tsuzuki Y. et al.Endothelins promote egg albumin-induced intestinal anaphylaxis in rats.Gastroenterology. 1998; 115: 348-356Abstract Full Text Full Text PDF PubMed Scopus (22) Google Scholar and pharmacological inhibition of the endothelin receptor ETA reportedly attenuated the magnitude of ovalbumin–induced intestinal anaphylaxis3Shigematsu T. Miura S. Hirokawa M. Hokari R. Higuchi H. Tsuzuki Y. et al.Endothelins promote egg albumin-induced intestinal anaphylaxis in rats.Gastroenterology. 1998; 115: 348-356Abstract Full Text Full Text PDF PubMed Scopus (22) Google Scholar or paw edema4Sampaio A.L. Rae G.A. D’Orleans-Juste P. Henriques M.G. ETA receptor antagonists inhibit allergic inflammation in the mouse.J Cardiovasc Pharmacol. 1995; 26: S416-S418PubMed Google Scholar in ovalbumin–sensitized animals. ET-1 can induce degranulation of MCs in mice,5Maurer M. Wedemeyer J. Metz M. Piliponsky A.M. Weller K. Chatterjea D. et al.Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (256) Google Scholar, 6Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google Scholar suggesting one mechanism by which increased levels of ET-1 might exacerbate anaphylaxis. However, the potential interactions between MC and ET-1 in anaphylaxis are complex. For example, mouse MCs can secrete ET-16Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google Scholar; MC-derived proteases, particularly MC-derived carboxypeptidase A,5Maurer M. Wedemeyer J. Metz M. Piliponsky A.M. Weller K. Chatterjea D. et al.Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (256) Google Scholar, 7Metz M. Piliponsky A.M. Chen C.C. Lammel V. Ǻbrink M. Pejler G. et al.Mast cells can enhance resistance to snake and honeybee venoms.Science. 2006; 313: 526-530Crossref PubMed Scopus (285) Google Scholar, 8Schneider L.A. Schlenner S.M. Feyerabend T.B. Wunderlin M. Rodewald H.R. Molecular mechanism of mast cell mediated innate defense against endothelin and snake venom sarafotoxin.J Exp Med. 2007; 204: 2629-2639Crossref PubMed Scopus (124) Google Scholar can hydrolyze ET-1 to a biologically inactive peptide by removing the C-terminal tryptophan8Schneider L.A. Schlenner S.M. Feyerabend T.B. Wunderlin M. Rodewald H.R. Molecular mechanism of mast cell mediated innate defense against endothelin and snake venom sarafotoxin.J Exp Med. 2007; 204: 2629-2639Crossref PubMed Scopus (124) Google Scholar; and pretreatment of in vitro–derived mouse MCs with ET-1 can significantly decrease the cells’ degranulation in response to IgE and antigen in vitro.6Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google Scholar In the present study we investigated whether the severity of IgE-dependent passive systemic or cutaneous anaphylaxis is altered in mice treated with BQ-123, an antagonist of the endothelin receptor ETA. MCs are thought to represent a critical effector cell in IgE-dependent anaphylaxis.1Summers C.W. Pumphrey R.S. Woods C.N. McDowell G. Pemberton P.W. Arkwright P.D. Factors predicting anaphylaxis to peanuts and tree nuts in patients referred to a specialist center.J Allergy Clin Immunol. 2008; 121 (e2): 632-638Abstract Full Text Full Text PDF PubMed Scopus (147) Google Scholar Prior functional assays and expression analyses of various MC populations demonstrated that ETA can be expressed by connective tissue–type MCs, including peritoneal MCs.5Maurer M. Wedemeyer J. Metz M. Piliponsky A.M. Weller K. Chatterjea D. et al.Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (256) Google Scholar, 6Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google Scholar, 9Metz M. Lammel V. Gibbs B.F. Maurer M. Inflammatory murine skin responses to UV-B light are partially dependent on endothelin-1 and mast cells.Am J Pathol. 2006; 169: 815-822Abstract Full Text Full Text PDF PubMed Scopus (46) Google Scholar However, immature bone marrow–derived cultured mouse mast cells (BMCMCs), which have some similarities to mucosal-type MCs, are largely unresponsive to ET-1 and have very low levels of receptor expression.6Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google Scholar We confirmed, using flow cytometric analysis, the substantial expression of ETA and weak expression of ETB on C57BL/6 mouse peritoneal MCs (Fig 1, A), as well as the absence of detectable surface ETB and very weak expression of ETA on C57BL/6 mouse BMCMCs (Fig 1, B). When BMCMCs were induced to undergo further maturation by maintaining the cells in the presence of IL-4 and stem cell factor, in addition to IL-3, the expression of ET receptors increased to levels approaching those of freshly isolated peritoneal MCs (Fig 1, C). Taken together, these results support the notion that a robust surface expression of ETA is a characteristic of mature murine MCs. Binding of ET-1 to ETA can induce strong and rapid degranulation of MCs,5Maurer M. Wedemeyer J. Metz M. Piliponsky A.M. Weller K. Chatterjea D. et al.Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (256) Google Scholar, 6Matsushima H. Yamada N. Matsue H. Shimada S. The effects of endothelin-1 on degranulation, cytokine, and growth factor production by skin-derived mast cells.Eur J Immunol. 2004; 34: 1910-1919Crossref PubMed Scopus (68) Google Scholar and ET-1 levels previously have been shown to be increased in certain animal models of anaphylaxis.2Sanchez-Cifuentes M.V. Rubio M.L. Ortega M. Largo R. Gomez-Garre M.D. Gonzalez Mangado N. et al.Endothelin-1 expression during early response after antigen challenge in brown-Norway rats.Pulm Pharmacol Ther. 1998; 11: 215-219Crossref PubMed Scopus (5) Google Scholar, 3Shigematsu T. Miura S. Hirokawa M. Hokari R. Higuchi H. Tsuzuki Y. et al.Endothelins promote egg albumin-induced intestinal anaphylaxis in rats.Gastroenterology. 1998; 115: 348-356Abstract Full Text Full Text PDF PubMed Scopus (22) Google Scholar We therefore hypothesized that ET-1 can have effects on MC-mediated allergic reactions. To investigate whether ET-1 can contribute to features of severe IgE- and MC-dependent allergic reactions, we first analyzed IgE-dependent passive systemic anaphylaxis in mice treated with the selective ETA antagonist BQ-123. Mice were sensitized intraperitoneally with IgE anti–2,4-dinitrophenol (DNP; in 100 μL of sterile, pyrogen-free 0.9% NaCl [saline]) and challenged intraperitoneally 24 hours later with 1 mg of DNP–human serum albumin (HSA; 30-40 DNP:1 HSA; Sigma, St Louis, Mo) in 100 μL of saline to induce anaphylaxis. Mice were pretreated with BQ-123 (Calbiochem, San Diego, Calif; 20 nmol in 200 μL of vehicle [saline + 0.5% dimethyl sulfoxide]) or 200 μL of vehicle intraperitoneally 45 minutes before challenge. Mice treated with BQ-123 exhibited a statistically significant, although modest, amelioration of the allergic reaction, as assessed by monitoring the ensuing change in body temperature (Fig 2, A). Furthermore, the reduced hypothermia was associated with a modest but statistically significant reduction in the extent of degranulation of peritoneal MCs, as assessed morphologically (Fig 2, B),5Maurer M. Wedemeyer J. Metz M. Piliponsky A.M. Weller K. Chatterjea D. et al.Mast cells promote homeostasis by limiting endothelin-1-induced toxicity.Nature. 2004; 432: 512-516Crossref PubMed Scopus (256) Google Scholar, 7Metz M. Piliponsky A.M. Chen C.C. Lammel V. Ǻbrink M. Pejler G. et al.Mast cells can enhance resistance to snake and honeybee venoms.Science. 2006; 313: 526-530Crossref PubMed Scopus (285) Google Scholar indicating that blockade of ETA results in decreased activation of MCs in this setting. In support of this conclusion, pretreatment with BQ-123 in mice subjected to passive systemic anaphylaxis also resulted in reduced levels of both MC-derived mouse MC protease 1 in the serum (Fig 2, C) and histamine in the peritoneal lavage fluid (Fig 2, D). To assess the possible contribution of ETA to IgE- and MC-dependent allergic responses in the skin, we next induced passive cutaneous anaphylaxis by means of intradermal injection of IgE anti-DNP (20 μg in 100 μL of saline) and then challenged the mice intraperitoneally with DNP-HSA (1 mg in 100 μL of saline) or 100 μL of saline alone 24 hours later. The increase in ear thickness and the extent of plasma extravasation were then quantified as measures of allergic inflammation. Injection of BQ-123 intravenously before elicitation of the allergic reaction again resulted in a significant reduction in the assessed parameters, with about 30% to 40% reduction in the measured responses compared with those seen in the identically sensitized and challenged but vehicle-treated mice (Fig 2, E and F). Taken together, our findings show that treatment with the ETA antagonist BQ-123 can ameliorate systemic or local IgE- and anaphylactic responses in mice. We hypothesize that one mechanism that might contribute to this observation is that the ETA antagonist can reduce ETA-dependent enhancement of MC activation by endogenous ET-1 present during the allergic reaction. AppendixExpanded legend for Fig 1Analysis of ET receptor expression on C57BL/6 mouse MCs. IL-3 in culture medium was derived from 20% WEHI-3–conditioned medium, IL-4 (from Calbiochem) was used at 30 ng/mL, and stem cell factor (from Amgen, Thousand Oaks, Calif) was used at 50 ng/mL. We assessed surface expression of ETA and ETB using rabbit anti-mouse ETA and ETB antibodies from Santa Cruz Biotechnology (Santa Cruz, Calif; M-60 and M-74, respectively).Expanded legend for Fig 2Elicitation and assessment of IgE-dependent anaphylactic reactions. A, Passive systemic anaphylaxis was induced by sensitizing C57BL/6 mice with 20 μg of IgE anti-DNP,E1Liu F.T. Bohn J.W. Ferry E.L. Yamamoto H. Molinaro C.A. Sherman L.A. et al.Monoclonal dinitrophenyl-specific murine IgE antibody: preparation, isolation, and characterization.J Immunol. 1980; 124: 2728-2737PubMed Google Scholar and mice were challenged 24 hours later intraperitoneally with 1 mg of DNP-HSA or vehicle. The selective ETA antagonist BQ-123 (20 nmol; Bachem, Bubendorf, Switzerland) in 200 μL of vehicle (saline [sterile, pyrogen-free 0.9% NaCl plus 0.5% dimethyl sulfoxide]) or vehicle alone was injected intraperitoneally 45 minutes before DNP-HSA or saline challenge. There were 9 mice per group pooled from 3 independent experiments, each of which produced similar results. B, Quantitative histomorphometry was performed on peritoneal lavage–derived MCs obtained 1 hour after DNP-HSA (30-40 DNP:1 HSA; Sigma) or saline challenge, with white bars, hatched bars, and black bars in Fig 2, B, indicating no, moderate, or extensive degranulation, respectively.E2Wershil B.K. Murakami T. Galli S.J. Mast cell-dependent amplification of an immunologically nonspecific inflammatory response. Mast cells are required for the full expression of cutaneous acute inflammation induced by phorbol 12- myristate 13-acetate.J Immunol. 1988; 140: 2356-2360PubMed Google Scholar There were 9 mice per group pooled from 3 independent experiments, each of which produced similar results. C and D, Levels of mouse MC protease 1 in the serum (Fig 2, C) were measured by means of ELISA (eBioscience, San Diego), and histamine concentrations in peritoneal lavage fluid (Fig 2, D) were measured with the Histareader 501 (RefLab, Copenhagen, Denmark), both according to the instructions of the manufacturer, 15 minutes after induction of passive systemic anaphylaxis. There were 9 to 10 (mouse MC protease 1) and 8 to 9 mice (histamine) per group pooled from 2 independent experiments, each of which produced similar results. Body temperature in the 2 treatment groups differed significantly 10 minutes after induction of passive systemic anaphylaxis (vehicle: −3.3°C ± 0.1°C vs BQ-123: −2.5°C ± 0.3°C; P < .01 unpaired Student t test, 2-tailed). E and F, Passive cutaneous anaphylaxis was induced in C57BL/6 mice by sensitizing them with 20 ng of IgE anti-DNP (in 20 μL of saline administered intradermally),E1Liu F.T. Bohn J.W. Ferry E.L. Yamamoto H. Molinaro C.A. Sherman L.A. et al.Monoclonal dinitrophenyl-specific murine IgE antibody: preparation, isolation, and characterization.J Immunol. 1980; 124: 2728-2737PubMed Google Scholar and control C57BL/6 mice were given 20 μL of saline administered intradermally. Mice were then challenged intraperitoneally 24 hours later with 200 μg of DNP-HSA (in 200 μL of saline plus 1% Evans blue). Mice were injected intraperitoneally with either BQ-123 (20 nmol in 200 μL administered intravenously) or vehicle 10 minutes before challenge, and ear swellingE3Wershil B.K. Mekori Y.A. Murakami T. Galli S.J. 125I-fibrin deposition in IgE- dependent immediate hypersensitivity reactions in mouse skin. Demonstration of the role of mast cells using genetically mast cell-deficient mice locally reconstituted with cultured mast cells.J Immunol. 1987; 139: 2605-2614PubMed Google Scholar (Fig 2, E) and extravasation of Evans blue dye (absorbance of the dye; Fig 2, F),E4Inagaki N. Nagai H. Koda A. Comparative study of IgG1 and IgE antibody mediated homologous PCA in the mouse ear. Lack of cross-desensitization of IgG1 antibody mediated PCA to IgE antibody mediated PCA.Int Arch Allergy Appl Immunol. 1988; 86: 325-330Crossref PubMed Scopus (13) Google Scholar each calculated as the percentage of the mean result in the vehicle-treated group, were measured 1 hour after DNP-HSA or saline challenge. There were 9 mice per group pooled from 3 independent experiments, each of which produced similar results. All results are presented as means ± SEMs (Fig 2, A) or means + SEMs (Fig 2, B-F). Expanded legend for Fig 1Analysis of ET receptor expression on C57BL/6 mouse MCs. IL-3 in culture medium was derived from 20% WEHI-3–conditioned medium, IL-4 (from Calbiochem) was used at 30 ng/mL, and stem cell factor (from Amgen, Thousand Oaks, Calif) was used at 50 ng/mL. We assessed surface expression of ETA and ETB using rabbit anti-mouse ETA and ETB antibodies from Santa Cruz Biotechnology (Santa Cruz, Calif; M-60 and M-74, respectively). Analysis of ET receptor expression on C57BL/6 mouse MCs. IL-3 in culture medium was derived from 20% WEHI-3–conditioned medium, IL-4 (from Calbiochem) was used at 30 ng/mL, and stem cell factor (from Amgen, Thousand Oaks, Calif) was used at 50 ng/mL. We assessed surface expression of ETA and ETB using rabbit anti-mouse ETA and ETB antibodies from Santa Cruz Biotechnology (Santa Cruz, Calif; M-60 and M-74, respectively). Expanded legend for Fig 2Elicitation and assessment of IgE-dependent anaphylactic reactions. A, Passive systemic anaphylaxis was induced by sensitizing C57BL/6 mice with 20 μg of IgE anti-DNP,E1Liu F.T. Bohn J.W. Ferry E.L. Yamamoto H. Molinaro C.A. Sherman L.A. et al.Monoclonal dinitrophenyl-specific murine IgE antibody: preparation, isolation, and characterization.J Immunol. 1980; 124: 2728-2737PubMed Google Scholar and mice were challenged 24 hours later intraperitoneally with 1 mg of DNP-HSA or vehicle. The selective ETA antagonist BQ-123 (20 nmol; Bachem, Bubendorf, Switzerland) in 200 μL of vehicle (saline [sterile, pyrogen-free 0.9% NaCl plus 0.5% dimethyl sulfoxide]) or vehicle alone was injected intraperitoneally 45 minutes before DNP-HSA or saline challenge. There were 9 mice per group pooled from 3 independent experiments, each of which produced similar results. B, Quantitative histomorphometry was performed on peritoneal lavage–derived MCs obtained 1 hour after DNP-HSA (30-40 DNP:1 HSA; Sigma) or saline challenge, with white bars, hatched bars, and black bars in Fig 2, B, indicating no, moderate, or extensive degranulation, respectively.E2Wershil B.K. Murakami T. Galli S.J. Mast cell-dependent amplification of an immunologically nonspecific inflammatory response. Mast cells are required for the full expression of cutaneous acute inflammation induced by phorbol 12- myristate 13-acetate.J Immunol. 1988; 140: 2356-2360PubMed Google Scholar There were 9 mice per group pooled from 3 independent experiments, each of which produced similar results. C and D, Levels of mouse MC protease 1 in the serum (Fig 2, C) were measured by means of ELISA (eBioscience, San Diego), and histamine concentrations in peritoneal lavage fluid (Fig 2, D) were measured with the Histareader 501 (RefLab, Copenhagen, Denmark), both according to the instructions of the manufacturer, 15 minutes after induction of passive systemic anaphylaxis. There were 9 to 10 (mouse MC protease 1) and 8 to 9 mice (histamine) per group pooled from 2 independent experiments, each of which produced similar results. Body temperature in the 2 treatment groups differed significantly 10 minutes after induction of passive systemic anaphylaxis (vehicle: −3.3°C ± 0.1°C vs BQ-123: −2.5°C ± 0.3°C; P < .01 unpaired Student t test, 2-tailed). E and F, Passive cutaneous anaphylaxis was induced in C57BL/6 mice by sensitizing them with 20 ng of IgE anti-DNP (in 20 μL of saline administered intradermally),E1Liu F.T. Bohn J.W. Ferry E.L. Yamamoto H. Molinaro C.A. Sherman L.A. et al.Monoclonal dinitrophenyl-specific murine IgE antibody: preparation, isolation, and characterization.J Immunol. 1980; 124: 2728-2737PubMed Google Scholar and control C57BL/6 mice were given 20 μL of saline administered intradermally. Mice were then challenged intraperitoneally 24 hours later with 200 μg of DNP-HSA (in 200 μL of saline plus 1% Evans blue). Mice were injected intraperitoneally with either BQ-123 (20 nmol in 200 μL administered intravenously) or vehicle 10 minutes before challenge, and ear swellingE3Wershil B.K. Mekori Y.A. Murakami T. Galli S.J. 125I-fibrin deposition in IgE- dependent immediate hypersensitivity reactions in mouse skin. Demonstration of the role of mast cells using genetically mast cell-deficient mice locally reconstituted with cultured mast cells.J Immunol. 1987; 139: 2605-2614PubMed Google Scholar (Fig 2, E) and extravasation of Evans blue dye (absorbance of the dye; Fig 2, F),E4Inagaki N. Nagai H. Koda A. Comparative study of IgG1 and IgE antibody mediated homologous PCA in the mouse ear. Lack of cross-desensitization of IgG1 antibody mediated PCA to IgE antibody mediated PCA.Int Arch Allergy Appl Immunol. 1988; 86: 325-330Crossref PubMed Scopus (13) Google Scholar each calculated as the percentage of the mean result in the vehicle-treated group, were measured 1 hour after DNP-HSA or saline challenge. There were 9 mice per group pooled from 3 independent experiments, each of which produced similar results. All results are presented as means ± SEMs (Fig 2, A) or means + SEMs (Fig 2, B-F). Elicitation and assessment of IgE-dependent anaphylactic reactions. A, Passive systemic anaphylaxis was induced by sensitizing C57BL/6 mice with 20 μg of IgE anti-DNP,E1Liu F.T. Bohn J.W. Ferry E.L. Yamamoto H. Molinaro C.A. Sherman L.A. et al.Monoclonal dinitrophenyl-specific murine IgE antibody: preparation, isolation, and characterization.J Immunol. 1980; 124: 2728-2737PubMed Google Scholar and mice were challenged 24 hours later intraperitoneally with 1 mg of DNP-HSA or vehicle. The selective ETA antagonist BQ-123 (20 nmol; Bachem, Bubendorf, Switzerland) in 200 μL of vehicle (saline [sterile, pyrogen-free 0.9% NaCl plus 0.5% dimethyl sulfoxide]) or vehicle alone was injected intraperitoneally 45 minutes before DNP-HSA or saline challenge. There were 9 mice per group pooled from 3 independent experiments, each of which produced similar results. B, Quantitative histomorphometry was performed on peritoneal lavage–derived MCs obtained 1 hour after DNP-HSA (30-40 DNP:1 HSA; Sigma) or saline challenge, with white bars, hatched bars, and black bars in Fig 2, B, indicating no, moderate, or extensive degranulation, respectively.E2Wershil B.K. Murakami T. Galli S.J. Mast cell-dependent amplification of an immunologically nonspecific inflammatory response. Mast cells are required for the full expression of cutaneous acute inflammation induced by phorbol 12- myristate 13-acetate.J Immunol. 1988; 140: 2356-2360PubMed Google Scholar There were 9 mice per group pooled from 3 independent experiments, each of which produced similar results. C and D, Levels of mouse MC protease 1 in the serum (Fig 2, C) were measured by means of ELISA (eBioscience, San Diego), and histamine concentrations in peritoneal lavage fluid (Fig 2, D) were measured with the Histareader 501 (RefLab, Copenhagen, Denmark), both according to the instructions of the manufacturer, 15 minutes after induction of passive systemic anaphylaxis. There were 9 to 10 (mouse MC protease 1) and 8 to 9 mice (histamine) per group pooled from 2 independent experiments, each of which produced similar results. Body temperature in the 2 treatment groups differed significantly 10 minutes after induction of passive systemic anaphylaxis (vehicle: −3.3°C ± 0.1°C vs BQ-123: −2.5°C ± 0.3°C; P < .01 unpaired Student t test, 2-tailed). E and F, Passive cutaneous anaphylaxis was induced in C57BL/6 mice by sensitizing them with 20 ng of IgE anti-DNP (in 20 μL of saline administered intradermally),E1Liu F.T. Bohn J.W. Ferry E.L. Yamamoto H. Molinaro C.A. Sherman L.A. et al.Monoclonal dinitrophenyl-specific murine IgE antibody: preparation, isolation, and characterization.J Immunol. 1980; 124: 2728-2737PubMed Google Scholar and control C57BL/6 mice were given 20 μL of saline administered intradermally. Mice were then challenged intraperitoneally 24 hours later with 200 μg of DNP-HSA (in 200 μL of saline plus 1% Evans blue). Mice were injected intraperitoneally with either BQ-123 (20 nmol in 200 μL administered intravenously) or vehicle 10 minutes before challenge, and ear swellingE3Wershil B.K. Mekori Y.A. Murakami T. Galli S.J. 125I-fibrin deposition in IgE- dependent immediate hypersensitivity reactions in mouse skin. Demonstration of the role of mast cells using genetically mast cell-deficient mice locally reconstituted with cultured mast cells.J Immunol. 1987; 139: 2605-2614PubMed Google Scholar (Fig 2, E) and extravasation of Evans blue dye (absorbance of the dye; Fig 2, F),E4Inagaki N. Nagai H. Koda A. Comparative study of IgG1 and IgE antibody mediated homologous PCA in the mouse ear. Lack of cross-desensitization of IgG1 antibody mediated PCA to IgE antibody mediated PCA.Int Arch Allergy Appl Immunol. 1988; 86: 325-330Crossref PubMed Scopus (13) Google Scholar each calculated as the percentage of the mean result in the vehicle-treated group, were measured 1 hour after DNP-HSA or saline challenge. There were 9 mice per group pooled from 3 independent experiments, each of which produced similar results. All results are presented as means ± SEMs (Fig 2, A) or means + SEMs (Fig 2, B-F).
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