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. 1994 Aug;82(4):509–515.

Regulation of synthesis of complement protein C4 in human fibroblasts: cell- and gene-specific effects of cytokines and lipopolysaccharide.

J Kulics 1, A Circolo 1, R C Strunk 1, H R Colten 1
PMCID: PMC1414926  PMID: 7835912

Abstract

Synthesis and secretion of the class III major histocompatibility complex (MHC) gene product, C4, were detected in human skin fibroblasts by metabolic labelling, immunoprecipitation and SDS-PAGE analysis. Pro-C4 (approximately 185,000 MW) was present in intracellular lysates, and the mature protein was present in extracellular media, with three bands of approximately 93,000, 75,000 and 33,000 MW, corresponding to the alpha, beta and gamma chains, respectively. C4 expression was increased in a dose-dependent manner by interferon-gamma (IFN-gamma), but was unaffected by interleukin-1 beta (IL-1 beta), IL-6 and tumor necrosis factor-alpha (TNF-alpha) alone, each of which augmented the expression of factor B, C3 and other complement proteins synthesized in fibroblasts. Simultaneous incubation of fibroblasts with IFN-gamma and TNF resulted in a synergistic increase in C4 synthesis. RNA blot analyses indicated that regulation of C4 synthesis by IFN-gamma and the combination of IFN-gamma and TNF was mediated primarily at a pretranslational level. Lipopolysaccharide (LPS) had no effect on C4 or HLA-DR synthesis in fibroblasts, either constitutive or IFN-gamma-regulated. These results are in contrast to the effects of LPS in monocytes, where LPS decreased constitutive synthesis and counter-regulated the IFN-gamma-enhanced expression of both C4 and HLA-DR. C2 expression in fibroblasts was also increased primarily by IFN-gamma. However, C2 synthesis was increased by LPS, 1L-1 and TNF, although to a lesser extent than the increase in synthesis of factor B stimulated by these mediators. These results show that up-regulation by IFN-gamma is a common feature of C2 and C4 expression in human cells that constitutively synthesize these proteins. In contrast, regulation of MHC class III and class II genes by LPS, TNF, IL-1, and IL-6 is cell- and gene-specific.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andoh A., Fujiyama Y., Bamba T., Hosoda S. Differential cytokine regulation of complement C3, C4, and factor B synthesis in human intestinal epithelial cell line, Caco-2. J Immunol. 1993 Oct 15;151(8):4239–4247. [PubMed] [Google Scholar]
  2. Barbey-Morel C., Pierce J. A., Campbell E. J., Perlmutter D. H. Lipopolysaccharide modulates the expression of alpha 1 proteinase inhibitor and other serine proteinase inhibitors in human monocytes and macrophages. J Exp Med. 1987 Oct 1;166(4):1041–1054. doi: 10.1084/jem.166.4.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carroll M. C., Belt T., Palsdottir A., Porter R. R. Structure and organization of the C4 genes. Philos Trans R Soc Lond B Biol Sci. 1984 Sep 6;306(1129):379–388. doi: 10.1098/rstb.1984.0098. [DOI] [PubMed] [Google Scholar]
  4. Carroll M. C., Campbell R. D., Bentley D. R., Porter R. R. A molecular map of the human major histocompatibility complex class III region linking complement genes C4, C2 and factor B. Nature. 1984 Jan 19;307(5948):237–241. doi: 10.1038/307237a0. [DOI] [PubMed] [Google Scholar]
  5. Chan A. C., Atkinson J. P. Functional studies on the secreted form of human C4 (C4s), two incompletely processed two-subunit C4 molecules (beta - alpha + gamma and beta + alpha - gamma), and pro-C4. J Immunol. 1984 Apr;132(4):1967–1971. [PubMed] [Google Scholar]
  6. Chan A. C., Atkinson J. P. Identification and structural characterization of two incompletely processed forms of the fourth component of human complement. J Clin Invest. 1983 Nov;72(5):1639–1649. doi: 10.1172/JCI111123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  8. Circolo A., Pierce G. F., Katz Y., Strunk R. C. Antiinflammatory effects of polypeptide growth factors. Platelet-derived growth factor, epidermal growth factor, and fibroblast growth factor inhibit the cytokine-induced expression of the alternative complement pathway activator factor B in human fibroblasts. J Biol Chem. 1990 Mar 25;265(9):5066–5071. [PubMed] [Google Scholar]
  9. Circolo A., Welgus H. G., Pierce G. F., Kramer J., Strunk R. C. Differential regulation of the expression of proteinases/antiproteinases in fibroblasts. Effects of interleukin-1 and platelet-derived growth factor. J Biol Chem. 1991 Jul 5;266(19):12283–12288. [PubMed] [Google Scholar]
  10. Cole F. S., Auerbach H. S., Goldberger G., Colten H. R. Tissue-specific pretranslational regulation of complement production in human mononuclear phagocytes. J Immunol. 1985 Apr;134(4):2610–2616. [PubMed] [Google Scholar]
  11. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  12. Goldberger G., Colten H. R. Precursor complement protein (pro-C4) is converted in vitro to native C4 by plasmin. Nature. 1980 Jul 31;286(5772):514–516. doi: 10.1038/286514a0. [DOI] [PubMed] [Google Scholar]
  13. Hall R. E., Colten H. R. Cell-free synthesis of the fourth component of guinea pig complement (C4): identification of a precursor of serum C4 (pro-C4). Proc Natl Acad Sci U S A. 1977 Apr;74(4):1707–1710. doi: 10.1073/pnas.74.4.1707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hamilton A. O., Jones L., Morrison L., Whaley K. Modulation of monocyte complement synthesis by interferons. Biochem J. 1987 Mar 15;242(3):809–815. doi: 10.1042/bj2420809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Harrison S. A., Mondino B. J., Mayer F. J. Scleral fibroblasts. Human leukocyte antigen expression and complement production. Invest Ophthalmol Vis Sci. 1990 Nov;31(11):2412–2419. [PubMed] [Google Scholar]
  16. Høgåsen A. K., Hestdal K., Abrahamsen T. G. Granulocyte-macrophage colony-stimulating factor, but not macrophage colony-stimulating factor, suppresses basal and lipopolysaccharide-stimulated complement factor production in human monocytes. J Immunol. 1993 Sep 15;151(6):3215–3224. [PubMed] [Google Scholar]
  17. Katz Y., Cole F. S., Strunk R. C. Synergism between gamma interferon and lipopolysaccharide for synthesis of factor B, but not C2, in human fibroblasts. J Exp Med. 1988 Jan 1;167(1):1–14. doi: 10.1084/jem.167.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Katz Y., Revel M., Strunk R. C. Interleukin 6 stimulates synthesis of complement proteins factor B and C3 in human skin fibroblasts. Eur J Immunol. 1989 Jun;19(6):983–988. doi: 10.1002/eji.1830190605. [DOI] [PubMed] [Google Scholar]
  19. Katz Y., Strunk R. C. IL-1 and tumor necrosis factor. Similarities and differences in stimulation of expression of alternative pathway of complement and IFN-beta 2/IL-6 genes in human fibroblasts. J Immunol. 1989 Jun 1;142(11):3862–3867. [PubMed] [Google Scholar]
  20. Katz Y., Strunk R. C. Synovial fibroblast-like cells synthesize seven proteins of the complement system. Arthritis Rheum. 1988 Nov;31(11):1365–1370. doi: 10.1002/art.1780311104. [DOI] [PubMed] [Google Scholar]
  21. Kaufman J. F., Auffray C., Korman A. J., Shackelford D. A., Strominger J. The class II molecules of the human and murine major histocompatibility complex. Cell. 1984 Jan;36(1):1–13. doi: 10.1016/0092-8674(84)90068-0. [DOI] [PubMed] [Google Scholar]
  22. Krakauer T., Oppenheim J. J. IL-1 and tumor necrosis factor-alpha each up-regulate both the expression of IFN-gamma receptors and enhance IFN-gamma-induced HLA-DR expression on human monocytes and a human monocytic cell line (THP-1). J Immunol. 1993 Feb 15;150(4):1205–1211. [PubMed] [Google Scholar]
  23. Kulics J., Colten H. R., Perlmutter D. H. Counterregulatory effects of interferon-gamma and endotoxin on expression of the human C4 genes. J Clin Invest. 1990 Mar;85(3):943–949. doi: 10.1172/JCI114523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  25. Levy W. P., Shively J., Rubinstein M., Del Valle U., Pestka S. Amino-terminal amino acid sequence of human leukocyte interferon. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5102–5104. doi: 10.1073/pnas.77.9.5102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Littman B. H., Dastvan F. F., Carlson P. L., Sanders K. M. Regulation of monocyte/macrophage C2 production and HLA-DR expression by IL-4 (BSF-1) and IFN-gamma. J Immunol. 1989 Jan 15;142(2):520–525. [PubMed] [Google Scholar]
  27. Maurer D. H., Hanke J. H., Mickelson E., Rich R. R., Pollack M. S. Differential presentation of HLA-DR, DQ, and DP restriction elements by interferon-gamma-treated dermal fibroblasts. J Immunol. 1987 Aug 1;139(3):715–723. [PubMed] [Google Scholar]
  28. Miura N., Prentice H. L., Schneider P. M., Perlmutter D. H. Synthesis and regulation of the two human complement C4 genes in stable transfected mouse fibroblasts. J Biol Chem. 1987 May 25;262(15):7298–7305. [PubMed] [Google Scholar]
  29. Moffat G. J., Lappin D., Birnie G. D., Whaley K. Complement biosynthesis in human synovial tissue. Clin Exp Immunol. 1989 Oct;78(1):54–60. [PMC free article] [PubMed] [Google Scholar]
  30. Morris K. M., Aden D. P., Knowles B. B., Colten H. R. Complement biosynthesis by the human hepatoma-derived cell line HepG2. J Clin Invest. 1982 Oct;70(4):906–913. doi: 10.1172/JCI110687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Perlmutter D. H., Cole F. S., Goldberger G., Colten H. R. Distinct primary translation products from human liver mRNA give rise to secreted and cell-associated forms of complement protein C2. J Biol Chem. 1984 Aug 25;259(16):10380–10385. [PubMed] [Google Scholar]
  32. Perlmutter D. H., Dinarello C. A., Punsal P. I., Colten H. R. Cachectin/tumor necrosis factor regulates hepatic acute-phase gene expression. J Clin Invest. 1986 Nov;78(5):1349–1354. doi: 10.1172/JCI112721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Perlmutter D. H., Goldberger G., Dinarello C. A., Mizel S. B., Colten H. R. Regulation of class III major histocompatibility complex gene products by interleukin-1. Science. 1986 May 16;232(4752):850–852. doi: 10.1126/science.3010455. [DOI] [PubMed] [Google Scholar]
  34. Reid K. B., Porter R. R. The proteolytic activation systems of complement. Annu Rev Biochem. 1981;50:433–464. doi: 10.1146/annurev.bi.50.070181.002245. [DOI] [PubMed] [Google Scholar]
  35. Reid K. B., Solomon E. Biosynthesis of the first component of complement by human fibroblasts. Biochem J. 1977 Dec 1;167(3):647–660. doi: 10.1042/bj1670647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sackstein R., Colten H. R. Molecular regulation of MHC class III (C4 and factor B) gene expression in mouse peritoneal macrophages. J Immunol. 1984 Sep;133(3):1618–1626. [PubMed] [Google Scholar]
  37. Sanceau J., Merlin G., Wietzerbin J. Tumor necrosis factor-alpha and IL-6 up-regulate IFN-gamma receptor gene expression in human monocytic THP-1 cells by transcriptional and post-transcriptional mechanisms. J Immunol. 1992 Sep 1;149(5):1671–1675. [PubMed] [Google Scholar]
  38. Stecher V. J., Thorbecke G. J. Sites of synthesis of serum proteins. 3. Production of beta-1-C, beta-1-E and transferrin by primate and rodent cell lines. J Immunol. 1967 Oct;99(4):660–668. [PubMed] [Google Scholar]
  39. Strunk R. C., Cole F. S., Perlmutter D. H., Colten H. R. gamma-Interferon increases expression of class III complement genes C2 and factor B in human monocytes and in murine fibroblasts transfected with human C2 and factor B genes. J Biol Chem. 1985 Dec 5;260(28):15280–15285. [PubMed] [Google Scholar]
  40. Whitehead A. S., Goldberger G., Woods D. E., Markham A. F., Colten H. R. Use of a cDNA clone for the fourth component of human complement (C4) for analysis of a genetic deficiency of C4 in guinea pig. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5387–5391. doi: 10.1073/pnas.80.17.5387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. de Ceulaer C., Papazoglou S., Whaley K. Increased biosynthesis of complement components by cultured monocytes, synovial fluid macrophages and skynovial membrane cells from patients with rheumatoid arthritis. Immunology. 1980 Sep;41(1):37–43. [PMC free article] [PubMed] [Google Scholar]

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