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Cytokine messenger RNA expression and proliferation status of intestinal mononuclear cells in noninflamed gut and Crohn's disease

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Abstract

T-cell activation and local cytokine production probably contribute to the pathogenesis of Crohn's disease. This study investigates the proliferative status of intestinal mononuclear cells (MNC) and cytokine messenger RNA (mRNA) production in gut tissue sections from patients with Crohn's disease and noninflamed controls. mRNA in situ hybridization was performed using 33P-labelled riboprobes for human interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, tumour necrosis factor-α and interferon-γ. The expression of the proliferation-associated antigen Ki-67 was analysed by immunohistochemical single and double staining. Compared with controls, where proliferation of MNC and cytokine expression was restricted to mucosal lymphoid follicles, inflamed gut tissue contained increased numbers of cells expressing cytokine mRNA, most prominently IL-1β and IL-6, but also interferon-γ and tumour necrosis factor-α. Proliferating T-cells were increased in number, and small amounts of IL-2-expressing cells were detected. IL-4 was expressed by a few cells exclusively in follicular germinal centres. IL-5 was negative. Proinflammatory cytokines are strongly expressed in situ in Crohn's disease and largely predominate over lymphokine mRNA. Our results provide in situ evidence of a local lymphocyte response in Crohn's disease with characteristics of a delayed-type hypersensitivity reaction.

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References

  1. Angerer LM, Cox KH, Angerer RC (1987) Demonstration of tissue-specific gene expression by in situ hybridization. Methods Enzymol 152:649–661

    Google Scholar 

  2. Autschbach F, Qiao L, Schürmann G, Wallich R, Moubayed P, Feller A, Meuer SC (in press) Immunohistochemical reactivity of adhesion structure workshop mAb (subpanels 2, 3, 4) in peripheral and mucosa associated lymphoid tissue and in Crohn's disease. In: Schlossman S, et al. (eds) Leucocyte typing V. Oxford University Press, Oxford

  3. Basham TY, Merigan TC (1983) Recombinant interferon-γ increases HLA-DR synthesis and expression. J Immunol 130:1492–1494

    Google Scholar 

  4. Bondi A, Chieregatti G, Eusebi V, Fulcheri E, Bussolati G (1982) The use of β-galactosidase as a tracer in immunocytochemistry. Histochemistry 76:153–158

    Google Scholar 

  5. Bosch FX, Leube RE, Achtstätter T, Moll R, Franke WW (1988) Expression of simple epithelial type cytokeratins in stratified epithelia as detected by immunolocalization and hybridization in situ. J Cell Biol 106:1635–1648

    Google Scholar 

  6. Breese E, Braegger CP, Corrigan CJ, Walker-Smith JA, MacDonald TT (1993) Interleukin-2 and interferon-γ-secreting T-cells in normal and diseased human intestinal mucosa. Immunology 78:127–131

    Google Scholar 

  7. Brynskov J, Tvede N, Andersen CB, Vilien M (1992) Increased concentrations of interleukin 1 β, interleukin-2, and soluble interleukin-2 receptors in endoscopical mucosal biopsy specimens with active inflammatory bowel disease. Gut 33:55–58

    Google Scholar 

  8. Capello M, Keshav S, Prince C, Jewell DP, Gordon S (1992) Detection for mRNAs for macrophage products in inflammatory bowel disease by in situ hybridization. Gut 33:1214–1219

    Google Scholar 

  9. Cominelli F, Nast CC, Dinarello CA, Gentilini P, Zipser RD (1984) Regulation of eicosanoid production in rabbit colon by interleukin 1. Gastroenterology 97:1400–1405

    Google Scholar 

  10. Cordell JL, Falini B, Erber WN, Ghosh AK, Abdulaziz Z, MacDonald S, Pulford KAF, Stein H, Mason DY (1984) Immunoenzymatic labelling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal antialkaline phosphatase (APAAP complexes). J Histochem Cytochem 32:219–229

    CAS  PubMed  Google Scholar 

  11. DeMaria R, Fais S, Silvestri M, Frati L, Pallone F, Santoni A, Testi R (1993) Continuous in vivo activation and transient hyporesponsiveness to TcR/CD3 triggering of human gut lamina propria lymphocytes. Eur J Immunol 23:3104–3108

    Google Scholar 

  12. Deusch K, Köhne G, Dähne I, Daum S, Classen M (1991) Mucosal immune regulation in Crohn's disease: pattern of cytokine synthesis in human intestinal lymphocytes. Gastroenterology 100 (II):A 573

  13. Fais S, Pallone F, Squarcia O, Biancone L, Ricci F, Paoluzi P, Boirivant M (1987) HLA-DR antigens on colonic epithelial cells in inflammatory bowel disease. I. Relation to the state of activation of lamina propria lymphocytes and to the epithelial expression of other surface markers. Clin Exp Immunol 68:605–612

    Google Scholar 

  14. Fais S, Capobianchi MR, Pallone F, DiMarco P, Boirivant M, Dianzani F, Torsoli A (1991) Spontaneous release of interferon γ by intestinal lamina propria lymphocytes in Crohn's disease. Kinetics of in vitro response to interferon γ inducers. Gut 32:403–407

    Google Scholar 

  15. Gerdes J, Lemke H, Baisch H, Wacker HH, Schwab U, Stein H (1984) Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J Immunol 133:1710–1715

    Google Scholar 

  16. Holzmann B, Johnson JP (1983) A beta-galactosidase linked immunoassay for the analysis of antigens on individual cells. J Immunol Methods 60:359–367

    Google Scholar 

  17. Hsu SM, Raine L, Fanger H (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabelled antibody (PAP) procedures. J Histochem Cytochem 29:577–580

    CAS  PubMed  Google Scholar 

  18. Isaacs KL, Sartor RB, Haskill S (1992) Cytokine messenger RNA profiles in inflammatory bowel disease mucosa detected by polymerase chain reaction amplification. Gastroenterology 103:1587–1595

    Google Scholar 

  19. Kusugami K, Matsuura T, West GA, Youngman KR, Rachmilewitz D, Fiocchi C (1991) Loss of interleukin-2-producing intestinal CD4+ T-cells in inflammatory bowel disease. Gastroenterology 101:1594–1605

    Google Scholar 

  20. Ligumski M, Simon PL, Karmeli F, Rachmilewitz D (1990) Role of interleukin 1 in inflammatory bowel disease — enhanced production during active disease. Gut 31:686–689

    Google Scholar 

  21. MacDonald TT, Hutchings P, Choy MY, Murch S, Cooke A (1990) Tumour necrosis factor-alpha and interferon-gamma production measured at the single cell level in normal and inflamed human intestine. Clin Exp Immunol 81:301–305

    Google Scholar 

  22. Mahida YR, Wu K, Jewell DP (1989). Enhanced production of interleukin 1-β by mononuclear cells isolated from mucosa with active ulcerative colitis or Crohn's disease. Gut 30:835–838

    Google Scholar 

  23. Malizia G, Calabrese A, Cottone M, Raimondo M, Trejdosiewicz LK, Smart CJ, Oliva L, Pagliaro L (1991) Expression of leucocyte adhesion molecules by mucosal mononuclear phagocytes in inflammatory bowel disease. Gastroenterology 100:150–159

    Google Scholar 

  24. Matsuura T, West GA, Youngman KR, Klein JS, Fiocchi C (1993) Immune activation genes in inflammatory bowel disease. Gastroenterology 104:448–458

    Google Scholar 

  25. Mazlam MZ, Hodgson HJF (1992) Peripheral blood monocyte cytokine production and acute phase response in inflammatory bowel disease. Gut 33:773–778

    Google Scholar 

  26. McCabe RP, Secrist H, Botney M, Egan M, Peters MG (1993) Cytokine mRNA expression in intestine from normal and inflammatory bowel disease patients. Clin Immunol Immunopathol 66:52–58

    Google Scholar 

  27. Mullin GE, Lazenby AJ, Harris ML, Bayless TM, James SP (1992) Increased interleukin-2 messenger RNA in the intestinal mucosal lesions of Crohn's disease but not ulcerative colitis. Gastroenterology 102:1620–1627

    Google Scholar 

  28. Mullin G, Maycon ZR, Sampat A, Vezza FR, Braun-Elwert L, Duchmann R, Weissman G, Katz S, McKinley M, James SP, Fisher SE (1993) Intestinal mucosal lymphokine production in Crohn's disease has a Th1 profile. Gastroenterology 104:A751

  29. Murch SH, Braegger CP, Walker-Smith JA, MacDonald TT (1993) Location of tumour necrosis factor α by immunohistochemistry in chronic inflammatory bowel disease. Gut 34:1705–1709

    Google Scholar 

  30. Nakamura M, Saito H, Kasanuki J, Tamura Y, Yoshida S (1992) Cytokine production in patients with inflammatory bowel disease. Gut 33:933–937

    Google Scholar 

  31. Nathan CF, Murray HW, Wiebe ME, Rubin BY (1983) Identification of interferon-gamma as the lymphokine that activates human macrophage oxidative metabolism and antimicrobial activity. J Exp Med 158:670–689

    Google Scholar 

  32. Nishida T, Nishina N, Takano M, Kawai K, Bando K, Masui Y, Nakai S, Hirai Y (1987) cDNA coning of IL-1-alpha and IL-1-beta from RNA from U937 cell line. Biochem Biophys Res Commun 143:345–352

    Google Scholar 

  33. Osborn L, Hession C, Tizard R, Vassallo C, Luhowskyj S, Chi-Rosso G, Lobb R (1990) Direct expression cloning of vascular cell adhesion molecule-1, a cytokine inducible endothelial protein that binds to lymphocytes. Cell 59:1203–1211

    Google Scholar 

  34. Ouyang Q, El-Youssef M, Yen-Lieberman B, Sapatnekar W, Youngman KR, Kusugami K, Fiocchi C (1988) Expression of HLA-DR antigens in inflammatory bowel disease mucosa: role of intestinal lamina propria mononuclear cell-derived interferon. Dig Dis Sci 33:1528–1536

    Google Scholar 

  35. Pallone F, Fais S, Squarcia O, Biancone L, Pozzilli P, Boirivant M (1987) Activation of peripheral blood and intestinal lamina propria lymphocytes in Crohn's disease. In vivo state of activation and in vitro response to stimulation as defined by the expression of early activation antigens. Gut 28:745–753

    Google Scholar 

  36. Qiao L, Schürmann G, Betzler M, Meuer SC (1991) Activation and signaling status of human lamina propria T lymphocytes. Gastroenterology 101:1529–1536

    Google Scholar 

  37. Qiao L, Schürmann G, Autschbach F, Wallich R, Meuer SC (1993) Human intestinal mucosa alters T-cell reactivities. Gastroenterology 105:814–819

    Google Scholar 

  38. Qiao L, Golling M, Autschbach F, Schürmann G, Meuer SC (1994) T cell receptor repertoire and mitotic responses of lamina propria T lymphocytes in inflammatory bowel disease. Clin Exp Immunol 97:303–308

    Google Scholar 

  39. Sartor RB (1994) Cytokines in intestinal inflammation: pathophysiological and clinical considerations. Gastroenterology 106:533–539

    Google Scholar 

  40. Springer TA (1990) Adhesion receptors of the immune system. Nature 346:425–433

    Google Scholar 

  41. Street NE, Mosman TR (1991) Functional diversity of T lymphocytes due to secretion of different cytokine patterns. FASEB J 5:171–177

    Google Scholar 

  42. Wang AM, Creasey AA, Ladner MB, Lin LS, Strickler J, VanArsdell JN, Arsdell JN, Yamamoto R, Mark DF (1985) Molecular cloning of the complementary DNA for human tumour necrosis factor. Science 228:149–154

    Google Scholar 

  43. Youngman KR, Simon PL, West GA, Cominelli F, Rachmilewitz D, Klein JS, Fiocchi C (1993) Localization of intestinal interleukin 1 activity and protein and gene expression to lamina propria cells. Gastroenterology 104:749–758

    Google Scholar 

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Autschbach, F., Qiao, L., Wallich, R. et al. Cytokine messenger RNA expression and proliferation status of intestinal mononuclear cells in noninflamed gut and Crohn's disease. Vichows Archiv A Pathol Anat 426, 51–60 (1995). https://doi.org/10.1007/BF00194698

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  • DOI: https://doi.org/10.1007/BF00194698

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