Inhibition of binding of interferon-gamma to its receptor by salicylates used in inflammatory bowel disease.

5-Aminosalicylic acid (5ASA), 4ASA, their N-acetylated metabolites N-acetyl-5ASA and N-acetyl-4ASA, olsalazine, and colchicine impair interferon-gamma (IFN gamma) induced HLA-DR expression on a colonic cell line, HT-29. The mechanism of this effect is now reported. HT-29 cells were cultured with 50 U/ml IFN gamma with or without drug, and northern blot analysis was performed using a probe for the beta chain of the DR molecule. IFN gamma led to a noticeable increase in HLA-DR mRNA which was attenuated by the drugs. Analysis of the specific binding of increasing concentrations of 125I-IFN gamma by non-linear regression showed a Kd of 1.35 x 10(-10) M and 2.3 x 10(5) binding sites per HT-29 cell. Binding of 125I-IFN gamma was reduced by incubation with increasing concentrations of unlabelled IFN gamma but not with IFN alpha. Incubation with therapeutic concentrations of drugs led to the following reductions in binding: 10 mM 5ASA, 20% (p < 0.001); 10 mM N-acetyl-5ASA, 24% (p < 0.01); 10 mM 4ASA, 21% (p < 0.005); 10 mM N-acetyl-4ASA, 29% (p < 0.001); and 1 mM olsalazine, 29% (p < 0.001). Colchicine (10(-7) M) and 10(-5) M prednisolone had no effect. Incubation with higher concentrations of the drugs revealed a dose-response effect on binding with complete inhibition by 100 mM 4ASA and 10 mM olsalazine, and lesser degrees of inhibition by 100 mM 5ASA, N-acetyl-5ASA, and N-acetyl-4ASA. At concentrations found in the rectal lumen, the salicylates used in inflammatory bowel disease impair the binding of IFN gamma to its receptor on colonic epithelial cells.

[1]  D. Jewell,et al.  Comparison of 5-aminosalicylic acid and N-acetylaminosalicylic acid uptake by the isolated human colonic epithelial cell. , 1992, Gut.

[2]  D. Jewell,et al.  Salicylates used in inflammatory bowel disease and colchicine impair interferon-gamma induced HLA-DR expression. , 1992, Gut.

[3]  C. Hawkey,et al.  5-Aminosalicylic acid is a potent inhibitor of interleukin 1 beta production in organ culture of colonic biopsy specimens from patients with inflammatory bowel disease. , 1991, Gut.

[4]  Timothy A. Springer,et al.  Adhesion receptors of the immune system , 1990, Nature.

[5]  H. Huitfeldt,et al.  Relation between T cell number and epithelial HLA class II expression quantified by image analysis in normal and inflamed human gastric mucosa. , 1990, Gut.

[6]  D. Jewell,et al.  Mechanism of action of 5-aminosalicylic acid and its derivatives. , 1990, Clinical science.

[7]  C. Benoist,et al.  Regulation of major histocompatibility complex class-II genes: X, Y and other letters of the alphabet. , 1990, Annual review of immunology.

[8]  A. O’Garra.  INTERLEUKINS AND THE IMMUNE SYSTEM 2 , 1989, The Lancet.

[9]  J. Madara,et al.  Interferon-gamma directly affects barrier function of cultured intestinal epithelial monolayers. , 1989, The Journal of clinical investigation.

[10]  R. Macdermott,et al.  Inhibition of antibody secretion by 5-aminosalicylic acid. , 1989, Gastroenterology.

[11]  M. Grisham,et al.  Sulfasalazine metabolites and dapsone attenuate formyl-methionyl-leucyl-phenylalanine-induced mucosal injury in rat ileum. , 1989, Gastroenterology.

[12]  R. Eliakim,et al.  Role of platelet-activating factor in ulcerative colitis. Enhanced production during active disease and inhibition by sulfasalazine and prednisolone. , 1988, Gastroenterology.

[13]  P. Brandtzaeg,et al.  Up‐regulation of the Expression of Secretory Component and HLA Molecules in a Human Colonic Cell Line by Tumour Necrosis Factor‐α and Gamma Interferon , 1988, Scandinavian journal of immunology.

[14]  P. Wesseling,et al.  Double‐blind comparison of 5‐aminosalicylic acid and acetyl‐5‐aminosalicylic acid suppositories in patients with idiopathic proctitis , 1988, Alimentary pharmacology & therapeutics.

[15]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[16]  D. Jewell,et al.  Class II antigen (HLA-DR) expression by intestinal epithelial cells in inflammatory diseases of colon. , 1987, Journal of clinical pathology.

[17]  S. Imamura,et al.  Effect of sulphasalazine and its metabolites on the generation of reactive oxygen species. , 1987, Gut.

[18]  E. Thorsby.  Structure and function of HLA molecules. , 1987, Transplantation proceedings.

[19]  S. Lee,et al.  Effects of interferon-gamma and tumor necrosis factor-alpha on the expression of an Ia antigen on a murine macrophage cell line. , 1986, Journal of immunology.

[20]  N. Cerf-Bensussan,et al.  Increased HLA-DR expression by enterocytes in children with celiac disease. , 1986, Gastroenterology.

[21]  M. Rubinstein,et al.  The interferon receptors. , 1986, CRC critical reviews in biochemistry.

[22]  K. Barrett,et al.  Inhibition of IgE-mediated mast cell degranulation by sulphasalazine. , 1985, European journal of pharmacology.

[23]  S. Fuggle,et al.  THE DETAILED DISTRIBUTION OF MHC CLASS II ANTIGENS IN NORMAL HUMAN ORGANS , 1984, Transplantation.

[24]  J. Rask-Madsen,et al.  Colonic azodisalicylate metabolism determined by in vivo dialysis in healthy volunteers and patients with ulcerative colitis. , 1984, Gastroenterology.

[25]  W. Stenson,et al.  Sulfasalazine inhibition of binding of N-formyl-methionyl-leucyl-phenylalanine (FMLP) to its receptor on human neutrophils. , 1984, Biochemical pharmacology.

[26]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[27]  D. Jewell,et al.  Expression of HLA-DR antigens by colonic epithelium in inflammatory bowel disease. , 1983, Clinical and experimental immunology.

[28]  C. Hawkey,et al.  Inhibition of prostaglandin synthetase in human rectal mucosa. , 1983, Gut.

[29]  E. Lobos,et al.  Sulfasalazine inhibits the synthesis of chemotactic lipids by neutrophils. , 1982, The Journal of clinical investigation.

[30]  M. Dallman,et al.  Graft-versus-host disease induces expression of Ia antigen in rat epidermal cells and gut epithelium , 1981, Nature.

[31]  H. Rogers,et al.  Pharmacokinetics of intravenous and oral prednisolone. , 1980, British journal of clinical pharmacology.

[32]  E. Thorsby,et al.  HLA‐DR‐like Antigens in the Epithelium of the Human Small Intestine , 1980, Scandinavian journal of immunology.

[33]  S. Truelove,et al.  The effect of topical N-acetyl-5-aminosalicylic acid in ulcerative colitis. , 1980, Scandinavian journal of gastroenterology.