Skip to main content
PMC home page
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1994 Aug;97(2):309–314. doi: 10.1111/j.1365-2249.1994.tb06086.x

Intraglomerular expression of transforming growth factor-beta 1 (TGF-beta 1) mRNA in patients with glomerulonephritis: quantitative analysis by competitive polymerase chain reaction.

M Iwano 1, Y Akai 1, Y Fujii 1, Y Dohi 1, N Matsumura 1, K Dohi 1
PMCID: PMC1534700  PMID: 8050182

Abstract

TGF-beta 1 is involved in the pathogenesis of glomerular sclerosis. We studied the intraglomerular expression of TGF-beta 1 mRNA in patients with glomerulonephritis using competitive polymerase chain reaction (PCR). This method is sensitive enough to quantify cDNA copies of mRNA present in small amounts of samples. Renal biopsy specimens were obtained from 42 patients with various kinds of glomerulonephritis. Ten glomeruli were dissected from renal biopsy specimens. Normal glomeruli were also obtained from the resected kidneys of eight patients with renal cell cancer. Total RNA was extracted from the glomeruli and reverse transcribed into cDNA with reverse transcriptase. To prepare samples containing identical amounts of beta-actin cDNA (8 pg), we performed competitive PCR by co-amplifying mutant templates of beta-actin with a unique EcoRI site. Next, to measure TGF-beta 1 cDNA, we performed competitive PCR by co-amplifying mutant templates of TGF-beta 1. We observed a higher glomerular expression of TGF-beta 1 mRNA in cases of mesangial proliferative glomerulonephritis having a moderate increase in mesangial matrix, diabetic nephropathy and diffuse proliferative lupus nephritis, compared with normal glomeruli. Results suggest that the intraglomerular synthesis of TGF-beta 1 may be involved in the progression of glomerulonephritis in humans.

Full text

PDF
310

Images in this article

310Fig. 1
on p.310
311Fig. 3
on p.311
312Fig. 4
on p.312

Selected References

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

  1. Bergstein J. M., Riley M., Bang N. U. Analysis of the plasminogen activator activity of the human glomerulus. Kidney Int. 1988 Apr;33(4):868–874. doi: 10.1038/ki.1988.78. [DOI] [PubMed] [Google Scholar]
  2. Border W. A., Noble N. A., Yamamoto T., Harper J. R., Yamaguchi Y. u., Pierschbacher M. D., Ruoslahti E. Natural inhibitor of transforming growth factor-beta protects against scarring in experimental kidney disease. Nature. 1992 Nov 26;360(6402):361–364. doi: 10.1038/360361a0. [DOI] [PubMed] [Google Scholar]
  3. Border W. A., Okuda S., Languino L. R., Ruoslahti E. Transforming growth factor-beta regulates production of proteoglycans by mesangial cells. Kidney Int. 1990 Feb;37(2):689–695. doi: 10.1038/ki.1990.35. [DOI] [PubMed] [Google Scholar]
  4. Border W. A., Okuda S., Languino L. R., Sporn M. B., Ruoslahti E. Suppression of experimental glomerulonephritis by antiserum against transforming growth factor beta 1. Nature. 1990 Jul 26;346(6282):371–374. doi: 10.1038/346371a0. [DOI] [PubMed] [Google Scholar]
  5. Border W. A., Ruoslahti E. Transforming growth factor-beta in disease: the dark side of tissue repair. J Clin Invest. 1992 Jul;90(1):1–7. doi: 10.1172/JCI115821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brenner B. M. Nephron adaptation to renal injury or ablation. Am J Physiol. 1985 Sep;249(3 Pt 2):F324–F337. doi: 10.1152/ajprenal.1985.249.3.F324. [DOI] [PubMed] [Google Scholar]
  7. Choi M. E., Kim E. G., Huang Q., Ballermann B. J. Rat mesangial cell hypertrophy in response to transforming growth factor-beta 1. Kidney Int. 1993 Nov;44(5):948–958. doi: 10.1038/ki.1993.336. [DOI] [PubMed] [Google Scholar]
  8. Coimbra T., Wiggins R., Noh J. W., Merritt S., Phan S. H. Transforming growth factor-beta production in anti-glomerular basement membrane disease in the rabbit. Am J Pathol. 1991 Jan;138(1):223–234. [PMC free article] [PubMed] [Google Scholar]
  9. Derynck R., Jarrett J. A., Chen E. Y., Eaton D. H., Bell J. R., Assoian R. K., Roberts A. B., Sporn M. B., Goeddel D. V. Human transforming growth factor-beta complementary DNA sequence and expression in normal and transformed cells. Nature. 1985 Aug 22;316(6030):701–705. doi: 10.1038/316701a0. [DOI] [PubMed] [Google Scholar]
  10. Edwards D. R., Murphy G., Reynolds J. J., Whitham S. E., Docherty A. J., Angel P., Heath J. K. Transforming growth factor beta modulates the expression of collagenase and metalloproteinase inhibitor. EMBO J. 1987 Jul;6(7):1899–1904. doi: 10.1002/j.1460-2075.1987.tb02449.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Feldman A. M., Ray P. E., Bristow M. R. Expression of alpha-subunits of G proteins in failing human heart: a reappraisal utilizing quantitative polymerase chain reaction. J Mol Cell Cardiol. 1991 Dec;23(12):1355–1358. doi: 10.1016/0022-2828(91)90182-l. [DOI] [PubMed] [Google Scholar]
  12. Floege J., Eng E., Young B. A., Alpers C. E., Barrett T. B., Bowen-Pope D. F., Johnson R. J. Infusion of platelet-derived growth factor or basic fibroblast growth factor induces selective glomerular mesangial cell proliferation and matrix accumulation in rats. J Clin Invest. 1993 Dec;92(6):2952–2962. doi: 10.1172/JCI116918. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Funk C. D., FitzGerald G. A. Eicosanoid forming enzyme mRNA in human tissues. Analysis by quantitative polymerase chain reaction. J Biol Chem. 1991 Jul 5;266(19):12508–12513. [PubMed] [Google Scholar]
  14. Gesualdo L., Pinzani M., Floriano J. J., Hassan M. O., Nagy N. U., Schena F. P., Emancipator S. N., Abboud H. E. Platelet-derived growth factor expression in mesangial proliferative glomerulonephritis. Lab Invest. 1991 Aug;65(2):160–167. [PubMed] [Google Scholar]
  15. Higuchi R., Krummel B., Saiki R. K. A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Res. 1988 Aug 11;16(15):7351–7367. doi: 10.1093/nar/16.15.7351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Horii Y., Muraguchi A., Iwano M., Matsuda T., Hirayama T., Yamada H., Fujii Y., Dohi K., Ishikawa H., Ohmoto Y. Involvement of IL-6 in mesangial proliferative glomerulonephritis. J Immunol. 1989 Dec 15;143(12):3949–3955. [PubMed] [Google Scholar]
  17. Isaka Y., Fujiwara Y., Ueda N., Kaneda Y., Kamada T., Imai E. Glomerulosclerosis induced by in vivo transfection of transforming growth factor-beta or platelet-derived growth factor gene into the rat kidney. J Clin Invest. 1993 Dec;92(6):2597–2601. doi: 10.1172/JCI116874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Johnson R. J., Raines E. W., Floege J., Yoshimura A., Pritzl P., Alpers C., Ross R. Inhibition of mesangial cell proliferation and matrix expansion in glomerulonephritis in the rat by antibody to platelet-derived growth factor. J Exp Med. 1992 May 1;175(5):1413–1416. doi: 10.1084/jem.175.5.1413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kaname S., Uchida S., Ogata E., Kurokawa K. Autocrine secretion of transforming growth factor-beta in cultured rat mesangial cells. Kidney Int. 1992 Dec;42(6):1319–1327. doi: 10.1038/ki.1992.423. [DOI] [PubMed] [Google Scholar]
  20. Laiho M., Saksela O., Keski-Oja J. Transforming growth factor-beta induction of type-1 plasminogen activator inhibitor. Pericellular deposition and sensitivity to exogenous urokinase. J Biol Chem. 1987 Dec 25;262(36):17467–17474. [PubMed] [Google Scholar]
  21. MacKay K., Striker L. J., Stauffer J. W., Doi T., Agodoa L. Y., Striker G. E. Transforming growth factor-beta. Murine glomerular receptors and responses of isolated glomerular cells. J Clin Invest. 1989 Apr;83(4):1160–1167. doi: 10.1172/JCI113996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nakamura T., Miller D., Ruoslahti E., Border W. A. Production of extracellular matrix by glomerular epithelial cells is regulated by transforming growth factor-beta 1. Kidney Int. 1992 May;41(5):1213–1221. doi: 10.1038/ki.1992.183. [DOI] [PubMed] [Google Scholar]
  23. Natori Y., O'Meara Y. M., Manning E. C., Minto A. W., Levine J. S., Weise W. J., Salant D. J. Production and polarized secretion of basement membrane components by glomerular epithelial cells. Am J Physiol. 1992 Jan;262(1 Pt 2):F131–F137. doi: 10.1152/ajprenal.1992.262.1.F131. [DOI] [PubMed] [Google Scholar]
  24. Okuda S., Languino L. R., Ruoslahti E., Border W. A. Elevated expression of transforming growth factor-beta and proteoglycan production in experimental glomerulonephritis. Possible role in expansion of the mesangial extracellular matrix. J Clin Invest. 1990 Aug;86(2):453–462. doi: 10.1172/JCI114731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Olson J. L., Heptinstall R. H. Nonimmunologic mechanisms of glomerular injury. Lab Invest. 1988 Nov;59(5):564–578. [PubMed] [Google Scholar]
  26. Peten E. P., Striker L. J., Carome M. A., Elliott S. J., Yang C. W., Striker G. E. The contribution of increased collagen synthesis to human glomerulosclerosis: a quantitative analysis of alpha 2IV collagen mRNA expression by competitive polymerase chain reaction. J Exp Med. 1992 Dec 1;176(6):1571–1576. doi: 10.1084/jem.176.6.1571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ponte P., Ng S. Y., Engel J., Gunning P., Kedes L. Evolutionary conservation in the untranslated regions of actin mRNAs: DNA sequence of a human beta-actin cDNA. Nucleic Acids Res. 1984 Feb 10;12(3):1687–1696. doi: 10.1093/nar/12.3.1687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Roberts A. B., Sporn M. B., Assoian R. K., Smith J. M., Roche N. S., Wakefield L. M., Heine U. I., Liotta L. A., Falanga V., Kehrl J. H. Transforming growth factor type beta: rapid induction of fibrosis and angiogenesis in vivo and stimulation of collagen formation in vitro. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4167–4171. doi: 10.1073/pnas.83.12.4167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sporn M. B., Roberts A. B. TGF-beta: problems and prospects. Cell Regul. 1990 Nov;1(12):875–882. doi: 10.1091/mbc.1.12.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Werber H. I., Emancipator S. N., Tykocinski M. L., Sedor J. R. The interleukin 1 gene is expressed by rat glomerular mesangial cells and is augmented in immune complex glomerulonephritis. J Immunol. 1987 May 15;138(10):3207–3212. [PubMed] [Google Scholar]
  31. Yamamoto T., Nakamura T., Noble N. A., Ruoslahti E., Border W. A. Expression of transforming growth factor beta is elevated in human and experimental diabetic nephropathy. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1814–1818. doi: 10.1073/pnas.90.5.1814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yoshioka K., Takemura T., Murakami K., Okada M., Hino S., Miyamoto H., Maki S. Transforming growth factor-beta protein and mRNA in glomeruli in normal and diseased human kidneys. Lab Invest. 1993 Feb;68(2):154–163. [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES