Abstract
Mechanisms leading to morphological changes of the small intestine during coeliac disease (CD) are not yet completely recognized; however, two main processes have been suggested recently: remodeling of mucosa by matrix metalloproteinases, and mucosal atrophy by apoptosis. The aim of this study was analysis of the expression of proteins regulating apoptosis in the small intestine of children with active CD (ACD) and potential CD (PCD). Jejunal biopsies of 43 children with PCD and untreated ACD and 21 control samples were analyzed by means of standard indirect immunohistochemical technique for Fas, Fas ligand (Fas-L), tissue transglutaminase (tTG), Bcl-2, and glutathione S-transferase (GST) expression. We found significantly lower numbers of Fas-expressing enterocytes in the ACD patients than in PCD patients and controls. Similarly, the number of Fas-positive mucosal lymphocytes was decreased in ACD when compared with PCD. The number of Fas-L- and tTG-expressing enterocytes and mucosal lymphocytes was higher in both PCD and ACD. On the other hand, the number of Bcl-2-positive mucosal lymphocytes in PCD as well as ACD was significantly lower. The expression of tTG in extracellular matrix was significantly higher in PCD and ACD when compared with controls. Our results showed that Fas and/or Fas-L, Bcl-2, and tTG may be involved in apoptotic pathways leading to mucosal atrophy in children with CD. tTG changes are in agreement with the presumed role of this protein in the pathogenesis of CD.
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Amendola A, Lombardi G, Oliverio S, et al. (1994) HIV-1 gp120-dependent induction of apoptosis in antigen-specific human T cell clones is characterized by "tissue" transglutaminase expression and prevented by cyclosporine A. FEBS Lett 339:258–264
Amendola A, Gougeon ML, Poccia F, et al. (1996) Induction of "tissue" transglutaminase in HIV pathogenesis: evidence for high rate of apoptosis of CD4+ T lymphocytes and accessory cells in lymphoid tissues. Proc Natl Acad Sci USA 93:11057–11062
Berke G (1995) The CTL's kiss of death. Cell 81:9-12
Brusco G, Muzi P, Ciccocioppo R, et al. (1999) Transglutaminase and coeliac disease: endomysial reactivity and small bowel expression. Clin Exp Immunol 118:371–375
Ciccocioppo R, Di Sabatino A, Gasbarrini G, et al. (1999) Apoptosis and gastrointestinal tract. Ital J Gastroenterol Hepatol 31:162–172
Ciccocioppo R, Di Sabatino A, Parroni R, et al. (2000) Cytolytic mechanisms of intraepithelial lymphocytes in coeliac disease (CoD). Clin Exp Immunol 120:235–240
Ciccocioppo R, Di Sabatino A, Parroni R, et al. (2000) Increased enterocyte apoptosis and Fas/Fas ligand system in celiac disease. Am J Clin Pathol 115:494–503
Clapper ML, Szarka CE (1998) Glutathione S-transferases: biomarkers of cancer risk and chemopreventive response. Chem Biol Interact 111:377–388
Di Sabatino A, Ciccocioppo R, D'Aló S, et al. (2001) Intraepithelial and lamina propria lymphocytes show distinct patterns of apoptosis whereas both populations are active in Fas based cytotoxicity in coeliac disease. Gut 49:380–386
Farrace MG, Picarelli A, Di Tola M, et al. (2001) Presence of anti-"tissue" transglutaminase antibodies in inflammatory intestinal diseases: an apoptosis-associated event? Cell Death Differ 8:767–770
Fassano A, Catassi C (2001) Current approaches to diagnosis and treatment of coeliac disease: an evolving spectrum. Gastroenterology 120:636–651
Feighery C (1999) Coeliac disease (clinical review). BMJ 319:236–239
Feldmann G, Haouzi D, Moreau A, et al. (2000) Opening of the mitochondrial permeability transition pore causes matrix expansion and outer membrane rupture in Fas-mediated hepatitis apoptosis in mice. Hepatology 31:674–683
Ferguson A, Arranz E, O'Mahony S (1993) Clinical and pathological spectrum of coeliac disease—active, silent, latent, potential. Gut 34:150–151
Fesus L, Thomazy V, Falus A (1987) Induction and activation of tissue transglutaminase during programmed cell death. FEBS Lett 224:104–108
Hall AG (1999) Glutathione and the regulation of cell death. Adv Exp Med Biol 457:199–203
Hall AG (1999) Review: The role of glutathione in the regulation of apoptosis. Eur J Clin Invest 29:238–245
Knight CR, Rees RC, Platts A, et al. (1993) Interleukin-2-activated human effector lymphocytes mediate cytotoxicity by inducing apoptosis in human leukaemia and solid tumour target cells. Immunology 79:535–541
Kroemer G, Petit P, Zamzami N, et al. (1995) The biochemistry of programmed cell death. FASEB J 9:1277–1287
Lebenthal E, Branski D (2002) Celiac disease: an emerging global problem. J Pediatr Gastroenterol Nutr 35:508–512
Maiuri L, Auricchio S, Coletta S, et al. (1998) Blockage of T-cell costimulation inhibits T-cell action in celiac disease. Gastroenterology 115:564–572
Maiuri L, Ciacci C, Raia V, et al. (2001) FAS engagement drives apoptosis of enterocytes of coeliac patients. Gut 48:418–424
Marsh MN, Growe PT (1995) Morphology of the mucosal lesion in gluten sensitivity. Baillieres Clin Gastroenterol 9:173–179
Martinez-Lorenzo MJ, Anel A, Gamen S, et al. (1999) Activated human T cells release bioactive Fas ligand and APO2 ligand in microvesicles. J Immunol 163:1274–1281
Marzari R, Sblattero D, Florian F, et al (2001) Molecular dissection of the tissue transglutaminase autoantibody response in celiac disease. J Immunol 166:4170–4176
Mastino A, Grelli S, Piacentini M, et al. (1993) Correlation between induction of lymphocyte apoptosis and prostaglandin E2 production by macrophages infected with HIV. Cell Immunol 152:120–130
Mitsiades N, Yu WH, Poulaki V, et al (2001) Matrix metalloproteinase-7-mediated cleavage of Fas ligand protects tumor cells from chemotherapeutic drug cytotoxicity. Cancer Res 61:577–581
Molberg O, McAdam SN, Korner R, et al. (1998) Tissue transglutaminase selectively modifies gliadin peptides that are recognized by gut-derived T cells in celiac disease. Nat Med 4:713–717
Molberg O, McAdam SN, Sollid LM (2000) Role of tissue transglutaminase in celiac disease. J Pediatr Gastroenterol Nutr 30:232–240
Moss SF, Attia L, Scholes JV, et al. (1996) Increased small intestinal apoptosis in coeliac disease. Gut 39:811–817
Mullighan CG, Bunce M, Welsh KI (1997) High resolution HLA-DQB1 typing using the polymerase chain reaction and sequence-specific primers. Tissue Antigens 50:688–692
Nagata S, Golstein P (1995) The Fas death factor. Science 267:1449–1456
Oberhuber G, Vogelsang H, Stolte M, et al (1996) Evidence that intestinal intraepithelial lymphocytes are activated cytotoxic T cells in celiac disease but not in giardiasis. Am J Pathol 148:1351–1357
Oberhuber G, Granditsch G, Vogelsang H (1999) The histopathology of coeliac disease: time for a standardized report scheme for pathologists. Eur J Gastroenterol Hepatol 11:1185–1194
Oliverio S, Amendola A, Rodolfo C, et al. (1999) Inhibition of "tissue" transglutaminase increases cell survival by preventing apoptosis. J Biol Chem 274:34123–34128
Piacentini M (1995) Tissue transglutaminase: a candidate effector element of physiological cell death. Curr Top Microbiol Immunol 200:163–175
Piredda L, Farrace MG, LoBello M, et al. (1999) Identification of "tissue" transglutaminase binding proteins in neural cells committed to apoptosis. FASEB J 13:355–364
Powell WC, Fingelton B, Wilson CL, et al. (1999) The metalloproteinase matrilysin proteolytically generates active soluble Fas ligand and potentiates epithelial cell apoptosis. Curr Biol 9:1441–1447
Quarsten H, Molberg O, Fugger L, et al. (1999) HLA binding and T cell recognition of a tissue transglutaminase-modified gliadin epitope. Eur J Immunol 29:2506–2514
Saas P, Walker PR, Hahne M, et al. (1997) Fas ligand expression by astrocytoma in vivo: maintaining immune privilege in the brain? J Clin Invest 99:1173–1178
Salmela MT, MacDonald TT, Black D, et al (2002) Upregulation of matrix metalloproteinases in a model of T cell-mediated tissue injury in the gut: analysis by gene array and in situ hybridization. Gut 51:540–547
Shan L, Molberg O, Parrot I, et al (2002) Structural basis for gluten intolerance in celiac sprue. Science 297:2275–2279
Shiner M, Eran M, Freier S, et al. (1998) Are intraepithelial lymphocytes in celiac mucosa responsible for inducing programmed cell death (apoptosis) in enterocytes? Histochemical demonstration of perforins in cytoplasmic granules of intraepithelial lymphocytes. J Pediatr Gastroenterol Nutr 27:393–396
Strater J, Walczak H, Hasel C, et al. (2001) CD95 ligand (CD95L) immunohistochemistry: a critical study on 12 antibodies. Cell Death Differ 8:273–278
Szondy Z, Molnar P, Nemes Z, et al. (1997) Differential expression of tissue transglutaminase during in vivo apoptosis of thymocytes induced via distinct signalling pathways. FEBS Lett 404:307–313
Thomazy V, Fesus L (1989) Differential expression of tissue transglutaminase in human cells. An immunohistochemical study. Cell Tissue Res 255:215–224
Troncone R, Greco L, Mayer M, et al. (1996) Latent and potential coeliac disease. Acta Paediatr (Suppl) 412:10–14
Uhlig H, Osman AA, Tanev ID, et al. (1998) Role of tissue transglutaminase in gliadin binding to reticular extracellular matrix and relation to coeliac disease autoantibodies. Autoimmunity 28:185–195
Van de Wal Y, Kooy Y, Van Veelen P, et al. (1998) Selective deamidation by tissue transglutaminase strongly enhances gliadin-specific T cell reactivity. J Immunol 161:1585–1588
Wagelie-Steffen AL, Hartmann K, Vliagoftis H, et al. (1998) Fas ligand (FasL, CD95L, APO-1L) expression in murine mast cells. Immunology 94:569–574
Walker Smith JA, Guandalini S, Schmitz J, et al. (1990) Revised criteria for diagnosis of coeliac disease. Arch Dis Child 65:909–911
Younes M, Schwartz MP, Finnie D, et al. (1999) Overexpression of Fas ligand (FasL) during malignant transformation in the large bowel and in Barrett's metaplasia of the esophagus. Hum Pathol 30:1309–1313
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We thank Mgr. Petra Řiháková for her technical assistance during preparation of the manuscript. This study was supported by grants IGA MZ CR NE 6197–3/2000 and MSM 151100001.
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Ehrmann, J., Kolek, A., Koďousek, R. et al. Immunohistochemical study of the apoptotic mechanisms in the intestinal mucosa during children's coeliac disease. Virchows Arch 442, 453–461 (2003). https://doi.org/10.1007/s00428-003-0794-2
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DOI: https://doi.org/10.1007/s00428-003-0794-2