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. 1995 Dec 1;182(6):1871–1881. doi: 10.1084/jem.182.6.1871

Release of the mucosal mast cell granule chymase, rat mast cell protease-II, during anaphylaxis is associated with the rapid development of paracellular permeability to macromolecules in rat jejunum

PMCID: PMC2192272  PMID: 7500033

Abstract

The soluble granule chymase, rat mast cell protease-II (RMCP-II), is abundantly expressed in intestinal mucosal mast cells (MMC) but its function is not known. One hypothesis is that RMCP-II degrades the epithelial basement membrane and promotes the loss of enterocytes typically associated with type I hypersensitivity reactions in the rat. To test this hypothesis more directly, ex vivo perfusion of the cranial mesenteric artery and jejunal lumen was used to monitor the anaphylactic release of RMCP-II and its effects on mucosal permeability and epithelial integrity. Within 2 min of intravascular challenge with soluble adult Nippostrongylus brasiliensis worm antigen there was a 1,000-fold (P < 0.02) increase in the concentration of RMCP-II in the vascular perfusate from the jejunum of Nippostrongylus-sensitized rats but not the controls. Similarly, translocation of RMCP-II into the gut lumen increased 10-fold (P < 0.02) after 2 min only in worm antigen- challenged immune rats. Using an identical protocol, but incorporating Evans blue-labeled human serum albumin (EB-HSA) in the vascular perfusate, the timing of the release of RMCP-II into the two compartments was very similar to the first experiment and furthermore the translocation of EB-HSA increased 18-fold (P < 0.05) after 4 min in sensitized rats challenged with worm antigen. To examine the effects of RMCP-II more directly 1 mg of the highly purified chymase was introduced into the cranial mesenteric artery in ex vivo perfused normal rats. A significant (P < 0.05) 70-fold increase in concentration of RMCP-II in jejunal perfusate occurred after 6 min. In a repeat dose- response experiment, infusion of 0.375, 0.75, or 1.5 mg of RMCP-II, together with EB-HSA, established that the cumulative amounts of RMCP- II and EB-HSA translocated from the vasculature to the gut lumen in each perfusion (during the 10-min period of RMCP-II infusion) were significantly correlated. Analysis of intestinal perfusates by SDS-PAGE and by Western blotting using monoclonal anti-RMCP-II antibody confirmed that there was a concomitant translocation of both the protease and EB-HSA into the gut lumen. Histological evaluation of the mucosa failed to reveal any significant morphological change in any of the experiments. The rapid development of macromolecular leak, its association with the translocation of RMCP-II, and the absence of gross epithelial lesions, suggest for the first time that a mast cell granule chymase increases epithelial permeability via a paracellular route and implies that the substrate may be a protein, or proteins, in the epithelial junctional complex.

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Selected References

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  1. Azghani A. O., Gray L. D., Johnson A. R. A bacterial protease perturbs the paracellular barrier function of transporting epithelial monolayers in culture. Infect Immun. 1993 Jun;61(6):2681–2686. doi: 10.1128/iai.61.6.2681-2686.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barrett K. E. Effect of histamine and other mast cell mediators on T84 epithelial cells. Ann N Y Acad Sci. 1992;664:222–231. doi: 10.1111/j.1749-6632.1992.tb39763.x. [DOI] [PubMed] [Google Scholar]
  3. Caplan M. S., Hedlund E., Hill N., MacKendrick W. The role of endogenous nitric oxide and platelet-activating factor in hypoxia-induced intestinal injury in rats. Gastroenterology. 1994 Feb;106(2):346–352. doi: 10.1016/0016-5085(94)90591-6. [DOI] [PubMed] [Google Scholar]
  4. Colgan S. P., Resnick M. B., Parkos C. A., Delp-Archer C., McGuirk D., Bacarra A. E., Weller P. F., Madara J. L. IL-4 directly modulates function of a model human intestinal epithelium. J Immunol. 1994 Sep 1;153(5):2122–2129. [PubMed] [Google Scholar]
  5. Crowe S. E., Perdue M. H. Gastrointestinal food hypersensitivity: basic mechanisms of pathophysiology. Gastroenterology. 1992 Sep;103(3):1075–1095. doi: 10.1016/0016-5085(92)90047-3. [DOI] [PubMed] [Google Scholar]
  6. Fernández-Gallardo S., Gijón M. A., García C., Furio V., Liu F. T., Sánchez Crespo M. The role of platelet-activating factor and peptidoleukotrienes in the vascular changes of rat passive anaphylaxis. Br J Pharmacol. 1992 Jan;105(1):119–125. doi: 10.1111/j.1476-5381.1992.tb14221.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Furuse M., Hirase T., Itoh M., Nagafuchi A., Yonemura S., Tsukita S., Tsukita S. Occludin: a novel integral membrane protein localizing at tight junctions. J Cell Biol. 1993 Dec;123(6 Pt 2):1777–1788. doi: 10.1083/jcb.123.6.1777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gibson S., Mackeller A., Newlands G. F., Miller H. R. Phenotypic expression of mast cell granule proteinases. Distribution of mast cell proteinases I and II in the rat digestive system. Immunology. 1987 Dec;62(4):621–627. [PMC free article] [PubMed] [Google Scholar]
  9. Gibson S., Miller H. R. Mast cell subsets in the rat distinguished immunohistochemically by their content of serine proteinases. Immunology. 1986 May;58(1):101–104. [PMC free article] [PubMed] [Google Scholar]
  10. Gonzalez-Crussi F., Hsueh W. Experimental model of ischemic bowel necrosis. The role of platelet-activating factor and endotoxin. Am J Pathol. 1983 Jul;112(1):127–135. [PMC free article] [PubMed] [Google Scholar]
  11. Harari Y., Russell D. A., Castro G. A. Anaphylaxis-mediated epithelial Cl- secretion and parasite rejection in rat intestine. J Immunol. 1987 Feb 15;138(4):1250–1255. [PubMed] [Google Scholar]
  12. Heyman M., Darmon N., Dupont C., Dugas B., Hirribaren A., Blaton M. A., Desjeux J. F. Mononuclear cells from infants allergic to cow's milk secrete tumor necrosis factor alpha, altering intestinal function. Gastroenterology. 1994 Jun;106(6):1514–1523. doi: 10.1016/0016-5085(94)90405-7. [DOI] [PubMed] [Google Scholar]
  13. Huntley J. F., Mackellar A., Miller H. R. Altered expression of mast cell proteases in the rat. Quantitative and immunohistochemical analysis of the distribution of rat mast cell proteases I and II during helminth infection. APMIS. 1993 Dec;101(12):953–962. doi: 10.1111/j.1699-0463.1993.tb00207.x. [DOI] [PubMed] [Google Scholar]
  14. Huntley J. F., Mackellar A., Newlands G. F., Irvine J., Miller H. R. Mapping of the rat mast cell granule proteinases RMCPI and II by enzyme-linked immunosorbent assay and paired immunofluorescence. APMIS. 1990 Oct;98(10):933–944. doi: 10.1111/j.1699-0463.1990.tb05018.x. [DOI] [PubMed] [Google Scholar]
  15. Kanwar S., Wallace J. L., Befus D., Kubes P. Nitric oxide synthesis inhibition increases epithelial permeability via mast cells. Am J Physiol. 1994 Feb;266(2 Pt 1):G222–G229. doi: 10.1152/ajpgi.1994.266.2.G222. [DOI] [PubMed] [Google Scholar]
  16. King S. J., Miller H. R. Anaphylactic release of mucosal mast cell protease and its relationship to gut permeability in Nippostrongylus-primed rats. Immunology. 1984 Apr;51(4):653–660. [PMC free article] [PubMed] [Google Scholar]
  17. Kubes P., Arfors K. E., Granger D. N. Platelet-activating factor-induced mucosal dysfunction: role of oxidants and granulocytes. Am J Physiol. 1991 Jun;260(6 Pt 1):G965–G971. doi: 10.1152/ajpgi.1991.260.6.G965. [DOI] [PubMed] [Google Scholar]
  18. MacNaughton W. K., Leach K. E., Prud'homme-Lalonde L., Ho W., Sharkey K. A. Ionizing radiation reduces neurally evoked electrolyte transport in rat ileum through a mast cell-dependent mechanism. Gastroenterology. 1994 Feb;106(2):324–335. doi: 10.1016/0016-5085(94)90589-4. [DOI] [PubMed] [Google Scholar]
  19. Madara J. L., Stafford J. Interferon-gamma directly affects barrier function of cultured intestinal epithelial monolayers. J Clin Invest. 1989 Feb;83(2):724–727. doi: 10.1172/JCI113938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Miller H. R., Huntley J. F., Newlands G. F., Irvine J. Granule chymases and the characterization of mast cell phenotype and function in rat and mouse. Monogr Allergy. 1990;27:1–30. [PubMed] [Google Scholar]
  21. Miller H. R., Jarrett W. F. Immune reactions in mucous membranes. I. Intestinal mast cell response during helminth expulsion in the rat. Immunology. 1971 Mar;20(3):277–288. [PMC free article] [PubMed] [Google Scholar]
  22. Miller H. R., Woodbury R. G., Huntley J. F., Newlands G. Systemic release of mucosal mast-cell protease in primed rats challenged with Nippostrongylus brasiliensis. Immunology. 1983 Jul;49(3):471–479. [PMC free article] [PubMed] [Google Scholar]
  23. Miller M. J., Zhang X. J., Barkemeyer B., Sadowska-Krowicka H., Eloby-Childress S., Gu X., Clark D. A. Potential role of histamine monochloramine in a rabbit model of ileitis. Scand J Gastroenterol. 1991 Aug;26(8):852–858. doi: 10.3109/00365529109037022. [DOI] [PubMed] [Google Scholar]
  24. Nawa Y., Miller H. R., Hall E., Jarrett E. E. Adoptive transfer of total and parasite-specific IgE responses in rats infected with Nippostrongylus brasiliensis. Immunology. 1981 Sep;44(1):119–123. [PMC free article] [PubMed] [Google Scholar]
  25. Nawa Y., Miller H. R. Protection against Nippostrongylus brasiliensis by adoptive immunization with immune thoracic duct lymphocytes. Cell Immunol. 1978 Apr;37(1):51–60. doi: 10.1016/0008-8749(78)90173-9. [DOI] [PubMed] [Google Scholar]
  27. Pemberton A. D., Miller H. R., John H. A., Scudamore C. L. Comparative studies of the Spi1 proteins of three equine alpha-1-proteinase inhibitor haplotypes following isolation by affinity chromatography. Int J Biochem. 1993 Sep;25(9):1263–1268. doi: 10.1016/0020-711x(93)90077-r. [DOI] [PubMed] [Google Scholar]
  28. Pennington A. M., Corpe C. P., Kellett G. L. Rapid regulation of rat jejunal glucose transport by insulin in a luminally and vascularly perfused preparation. J Physiol. 1994 Jul 15;478(Pt 2):187–193. doi: 10.1113/jphysiol.1994.sp020241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Perdue M. H., Kosecka U., Crowe S. Antigen-mediated effects on epithelial function. Ann N Y Acad Sci. 1992;664:325–334. doi: 10.1111/j.1749-6632.1992.tb39771.x. [DOI] [PubMed] [Google Scholar]
  30. Perdue M. H., Masson S., Wershil B. K., Galli S. J. Role of mast cells in ion transport abnormalities associated with intestinal anaphylaxis. Correction of the diminished secretory response in genetically mast cell-deficient W/Wv mice by bone marrow transplantation. J Clin Invest. 1991 Feb;87(2):687–693. doi: 10.1172/JCI115047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Persson C. G., Gustafsson B., Erjefält J. S., Sundler F. Mucosal exudation of plasma is a noninjurious intestinal defense mechanism. Allergy. 1993 Nov;48(8):581–586. doi: 10.1111/j.1398-9995.1993.tb00752.x. [DOI] [PubMed] [Google Scholar]
  32. Pirie-Shepherd S. R., Miller H. R., Ryle A. Differential inhibition of rat mast cell proteinase I and II by members of the alpha-1-proteinase inhibitor family of serine proteinase inhibitors. J Biol Chem. 1991 Sep 15;266(26):17314–17319. [PubMed] [Google Scholar]
  33. Rippe B., Haraldsson B. Transport of macromolecules across microvascular walls: the two-pore theory. Physiol Rev. 1994 Jan;74(1):163–219. doi: 10.1152/physrev.1994.74.1.163. [DOI] [PubMed] [Google Scholar]
  34. Sage H., Woodbury R. G., Bornstein P. Structural studies on human type IV collagen. J Biol Chem. 1979 Oct 10;254(19):9893–9900. [PubMed] [Google Scholar]
  35. Suzuki K., Lees M., Newlands G. F., Nagase H., Woolley D. E. Activation of precursors for matrix metalloproteinases 1 (interstitial collagenase) and 3 (stromelysin) by rat mast-cell proteinases I and II. Biochem J. 1995 Jan 1;305(Pt 1):301–306. doi: 10.1042/bj3050301. [DOI] [PMC free article] [PubMed] [Google Scholar]

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