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PMID: 9502421 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Constitutive transcription of the human interleukin-6 gene by rheumatoid synoviocytes: spontaneous activation of NF-kappaB and CBF1.

The American journal of pathology ·Vol. 152 ·No. 3 ·1998-03-00 ·Pages 793-803

Miyazawa K, Mori A, Yamamoto K, Okudaira H

Abstract

The involvement of IL-6 in the pathogenesis of rheumatoid arthritis (RA) has been recently demonstrated. In the present study, we investigated the cellular and molecular mechanisms involved in the spontaneous IL-6 production by the fibroblast-like synoviocytes (FLSs) obtained from patients with RA. Cloned FLSs were established from the bulk cultures of FLSs by the limiting dilution method. Some FLS clones spontaneously produced large amounts of IL-6, whereas others produced low amounts of it. Neither anti-human TNF-alpha nor IL-1 antibody affected spontaneous IL-6 production of these FLS clones, suggesting that IL-6 production of the FLSs was endogenously up-regulated. A luciferase reporter plasmid containing the human IL-6 promoter region was significantly transcribed when transfected into the IL-6 high-producing clones, indicating that the rheumatoid FLSs retained constitutive transcriptional activity of the IL-6 gene. Electrophoretic mobility shift assays revealed that the binding activity of p50 and p65 NF-kappaB subunits and CBF1 was significantly enhanced in the IL-6 high-producing clones compared with that of IL-6 low-producing clones and cultured sarcoma cells, suggesting that spontaneous activation of NF-kappaB and CBF1 may lead to the constitutive transcription of the IL-6 gene by rheumatoid FLSs.

MeSH Terms
Arthritis, Rheumatoid/metabolism Blotting, Northern Cells, Cultured Clone Cells DNA Probes/chemistry DNA-Binding Proteins/biosynthesis Dexamethasone/pharmacology Humans Interleukin-1/pharmacology Interleukin-6/biosynthesis,genetics NF-kappa B/biosynthesis Neoplasm Proteins Osteoarthritis/metabolism Synovial Membrane/drug effects,metabolism Transcription Factors/biosynthesis Transcription, Genetic Transfection Tumor Necrosis Factor-alpha/pharmacology
Chemicals
DNA Probes DNA-Binding Proteins Interleukin-1 Interleukin-6 NF-kappa B Neoplasm Proteins Transcription Factors Tumor Necrosis Factor-alpha Dexamethasone
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Miyazawa K
Department of Medicine and Physical Therapy, Faculty of Medicine, University of Tokyo, Japan.
Mori A
Yamamoto K
Okudaira H
References (64)
64 references, click to expand
  1. Interleukin-1 and tumour necrosis factor mRNA expression in rheumatoid arthritis: prolonged production of IL-1 alpha.
    Clin Exp Immunol. 1988 Sep;73(3):449-55 PMID: 3264773
  2. Anchorage-independent growth of synoviocytes from arthritic and normal joints. Stimulation by exogenous platelet-derived growth factor and inhibition by transforming growth factor-beta and retinoids.
    J Clin Invest. 1989 Apr;83(4):1267-76 PMID: 2784799
  3. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells.
    Nucleic Acids Res. 1989 Aug 11;17(15):6419 PMID: 2771659
  4. A multiple cytokine- and second messenger-responsive element in the enhancer of the human interleukin-6 gene: similarities with c-fos gene regulation.
    Mol Cell Biol. 1989 Dec;9(12):5537-47 PMID: 2511437
  5. A protein binding to the J kappa recombination sequence of immunoglobulin genes contains a sequence related to the integrase motif.
    Nature. 1989 Dec 21-28;342(6252):934-7 PMID: 2556644
  6. Involvement of a NF-kappa B-like transcription factor in the activation of the interleukin-6 gene by inflammatory lymphokines.
    Mol Cell Biol. 1990 Feb;10(2):561-8 PMID: 2405250
  7. Rheumatoid arthritis. Pathophysiology and implications for therapy.
    N Engl J Med. 1990 May 3;322(18):1277-89 PMID: 2271017
  8. Quantitative analysis of cytokine gene expression in rheumatoid arthritis.
    J Immunol. 1990 May 1;144(9):3347-53 PMID: 2109776
  9. IL-6 functions as an exocrine hormone in inflammation. Hepatocytes undergoing acute phase responses require exogenous IL-6.
    J Immunol. 1990 May 15;144(10):3804-8 PMID: 2110214
  10. Constitutive and interleukin-1 (IL-1)-inducible factors interact with the IL-1-responsive element in the IL-6 gene.
    Mol Cell Biol. 1990 Jun;10(6):2757-64 PMID: 2111442
  11. Interleukin-6 induction by tumor necrosis factor and interleukin-1 in human fibroblasts involves activation of a nuclear factor binding to a kappa B-like sequence.
    Mol Cell Biol. 1990 Jul;10(7):3818-23 PMID: 2192263
  12. Detection of interleukin 8 biological activity in synovial fluids from patients with rheumatoid arthritis and production of interleukin 8 mRNA by isolated synovial cells.
    Eur J Immunol. 1990 Sep;20(9):2141-4 PMID: 2209707
  13. Selective attraction of monocytes and T lymphocytes of the memory phenotype by cytokine RANTES.
    Nature. 1990 Oct 18;347(6294):669-71 PMID: 1699135
  14. Constitutive production of inflammatory and mitogenic cytokines by rheumatoid synovial fibroblasts.
    J Exp Med. 1991 Mar 1;173(3):569-74 PMID: 1997647
  15. The p65 subunit is responsible for the strong transcription activating potential of NF-kappa B.
    EMBO J. 1991 Dec;10(12):3805-17 PMID: 1935902
  16. Engagement of major histocompatibility complex class II molecules by superantigen induces inflammatory cytokine gene expression in human rheumatoid fibroblast-like synoviocytes.
    J Exp Med. 1992 Feb 1;175(2):613-6 PMID: 1732419
  17. Functional characterization of the NF-kappa B p65 transcriptional activator and an alternatively spliced derivative.
    Mol Cell Biol. 1992 Feb;12(2):444-54 PMID: 1732726
  18. The 65-kDa subunit of human NF-kappa B functions as a potent transcriptional activator and a target for v-Rel-mediated repression.
    Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1875-9 PMID: 1542686
  19. Interleukin 6 levels in synovial fluids of patients with different arthritides: correlation with local IgM rheumatoid factor and systemic acute phase protein production.
    J Rheumatol. 1992 Jan;19(1):63-8 PMID: 1556702
  20. The importance of the T cell in initiating and maintaining the chronic synovitis of rheumatoid arthritis.
    Arthritis Rheum. 1992 Jul;35(7):729-35 PMID: 1622409
  21. Treatment of severe rheumatoid arthritis by anti-interleukin 6 monoclonal antibody.
    J Rheumatol. 1993 Feb;20(2):259-62 PMID: 8474061
  22. Interleukin-6 in biology and medicine.
    Adv Immunol. 1993;54:1-78 PMID: 8379461
  23. The Epstein-Barr virus immortalizing protein EBNA-2 is targeted to DNA by a cellular enhancer-binding protein.
    Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9237-41 PMID: 8415684
  24. Physical association and functional antagonism between the p65 subunit of transcription factor NF-kappa B and the glucocorticoid receptor.
    Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):752-6 PMID: 8290595
  25. Local proliferation of fibroblast-like synoviocytes contributes to synovial hyperplasia. Results of proliferating cell nuclear antigen/cyclin, c-myc, and nucleolar organizer region staining.
    Arthritis Rheum. 1994 Feb;37(2):212-20 PMID: 7510485
  26. Multiple regulatory elements in the interleukin-6 gene mediate induction by prostaglandins, cyclic AMP, and lipopolysaccharide.
    Mol Cell Biol. 1994 Jul;14(7):4443-54 PMID: 8007951
  27. Mediation of Epstein-Barr virus EBNA2 transactivation by recombination signal-binding protein J kappa.
    Science. 1994 Jul 1;265(5168):92-5 PMID: 8016657
  28. Synovium as a source of interleukin 6 in vitro. Contribution to local and systemic manifestations of arthritis.
    J Clin Invest. 1989 Feb;83(2):585-92 PMID: 2464001
  29. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  30. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene.
    Nucleic Acids Res. 1985 Apr 11;13(7):2485-502 PMID: 2987855
  31. Purification to homogeneity and characterization of human B-cell differentiation factor (BCDF or BSFp-2).
    Proc Natl Acad Sci U S A. 1985 Aug;82(16):5490-4 PMID: 2410927
  32. Induction of a 26-kDa-protein mRNA in human cells treated with an interleukin-1-related, leukocyte-derived factor.
    Eur J Biochem. 1985 Oct 15;152(2):253-7 PMID: 3876934
  33. Human recombinant interleukin 1 stimulates collagenase and prostaglandin E2 production by human synovial cells.
    J Clin Invest. 1986 Feb;77(2):645-8 PMID: 3003163
  34. Development of criteria for the classification and reporting of osteoarthritis. Classification of osteoarthritis of the knee. Diagnostic and Therapeutic Criteria Committee of the American Rheumatism Association.
    Arthritis Rheum. 1986 Aug;29(8):1039-49 PMID: 3741515
  35. Interleukin 1 and poly(rI).poly(rC) induce production of a hybridoma growth factor by human fibroblasts.
    Eur J Immunol. 1987 Jan;17(1):1-7 PMID: 3545852
  36. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
    Anal Biochem. 1987 Apr;162(1):156-9 PMID: 2440339
  37. Spontaneous production of an interleukin 1-like factor by cloned rheumatoid synovial cells in long-term culture.
    J Clin Invest. 1987 Sep;80(3):786-96 PMID: 2442197
  38. Interferon beta 2/B-cell stimulatory factor type 2 shares identity with monocyte-derived hepatocyte-stimulating factor and regulates the major acute phase protein response in liver cells.
    Proc Natl Acad Sci U S A. 1987 Oct;84(20):7251-5 PMID: 2444978
  39. Elevated cytokine levels in synovial fluid of rheumatoid arthritis correlates with the presence of cytomegalovirus genome.
    Autoimmunity. 1994;17(4):333-7 PMID: 7948616
  40. Transcriptional regulation of the human interleukin-6 gene promoter in human T-cell leukemia virus type I-infected T-cell lines: evidence for the involvement of NF-kappa B.
    Blood. 1994 Nov 1;84(9):2904-11 PMID: 7949164
  41. Inhibition of the production and effects of interleukin-1 and tumor necrosis factor alpha in rheumatoid arthritis.
    Arthritis Rheum. 1995 Feb;38(2):151-60 PMID: 7848304
  42. Activation of monocyte effector genes and STAT family transcription factors by inflammatory synovial fluid is independent of interferon gamma.
    J Exp Med. 1995 Mar 1;181(3):1015-25 PMID: 7869026
  43. Interleukin-6 in relation to other proinflammatory cytokines, chemotactic activity and neutrophil activation in rheumatoid synovial fluid.
    Ann Rheum Dis. 1995 Jan;54(1):33-8 PMID: 7880119
  44. Interleukin-6 (IL-6) induces the proliferation of synovial fibroblastic cells in the presence of soluble IL-6 receptor.
    Br J Rheumatol. 1995 Apr;34(4):321-5 PMID: 7788145
  45. Interleukin-6 family of cytokines and gp130.
    Blood. 1995 Aug 15;86(4):1243-54 PMID: 7632928
  46. Regulation of interleukin-6 gene expression by steroids.
    Ann N Y Acad Sci. 1995 Jul 21;762:79-87; discussion 87-8 PMID: 7668576
  47. Monoclonal anti-TNF alpha antibody as a probe of pathogenesis and therapy of rheumatoid disease.
    Immunol Rev. 1995 Apr;144:195-223 PMID: 7590814
  48. Nuclear factor-kappa B in rheumatoid synovium. Localization of p50 and p65.
    Arthritis Rheum. 1995 Dec;38(12):1762-70 PMID: 8849348
  49. Distinct roles of the two tumor necrosis factor (TNF) receptors in modulating TNF and lymphotoxin alpha effects.
    J Biol Chem. 1996 Apr 19;271(16):9778-84 PMID: 8621658
  50. Constitutively active human Notch1 binds to the transcription factor CBF1 and stimulates transcription through a promoter containing a CBF1-responsive element.
    Proc Natl Acad Sci U S A. 1996 May 28;93(11):5663-7 PMID: 8643633
  51. Activation of the transcription factor nuclear factor-kappaB in human inflamed synovial tissue.
    Arthritis Rheum. 1996 Apr;39(4):583-91 PMID: 8630106
  52. Role of cytokines in rheumatoid arthritis.
    Annu Rev Immunol. 1996;14:397-440 PMID: 8717520
  53. Invasive fibroblast-like synoviocytes in rheumatoid arthritis. Passive responders or transformed aggressors?
    Arthritis Rheum. 1996 Nov;39(11):1781-90 PMID: 8912499
  54. The mammalian transcriptional repressor RBP (CBF1) regulates interleukin-6 gene expression.
    Mol Cell Biol. 1997 Jan;17(1):1-9 PMID: 8972179
  55. Structure and expression of human B cell stimulatory factor-2 (BSF-2/IL-6) gene.
    EMBO J. 1987 Oct;6(10):2939-45 PMID: 3500852
  56. Transcriptional regulation of the interferon-beta 2/B cell differentiation factor BSF-2/hepatocyte-stimulating factor gene in human fibroblasts by other cytokines.
    J Immunol. 1988 Feb 1;140(3):974-7 PMID: 3276786
  57. Detection of activated T cell products in the rheumatoid joint using cDNA probes to Interleukin-2 (IL-2) IL-2 receptor and IFN-gamma.
    Clin Exp Immunol. 1988 Feb;71(2):295-301 PMID: 3127092
  58. B cell stimulating factor 2/interleukin 6 is a costimulant for human thymocytes and T lymphocytes.
    J Exp Med. 1988 Mar 1;167(3):1253-8 PMID: 3127525
  59. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis.
    Arthritis Rheum. 1988 Mar;31(3):315-24 PMID: 3358796
  60. Synergism between interleukin-6 and interleukin-3 in supporting proliferation of human hematopoietic stem cells: comparison with interleukin-1 alpha.
    Blood. 1988 Jun;71(6):1759-63 PMID: 3259443
  61. Interleukin-6 in synovial fluid and serum of patients with rheumatoid arthritis and other inflammatory arthritides.
    Arthritis Rheum. 1988 Jun;31(6):784-8 PMID: 3260102
  62. Interleukin 4 is at 5q31 and interleukin 6 is at 7p15.
    Hum Genet. 1988 Aug;79(4):335-7 PMID: 3261707
  63. Genomic structure of the murine IL-6 gene. High degree conservation of potential regulatory sequences between mouse and human.
    J Immunol. 1988 Dec 1;141(11):3875-81 PMID: 3263439
  64. Excessive production of interleukin 6/B cell stimulatory factor-2 in rheumatoid arthritis.
    Eur J Immunol. 1988 Nov;18(11):1797-801 PMID: 2462501
Article Info
Journal
The American journal of pathology
Abbr.
Am J Pathol
ISSN
0002-9440
Published
1998-03-00
Pages
793-803
Language
English
Region
United States
NLM ID
0370502
PMCID
PMC1858393
Subset
IM
Corrections
CommentIn
PMID: 9502403
External Links
PubMed source PMC Full Text View journal details
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