Skip to main content
PMC home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1995 Dec;147(6):1759–1769.

Immunolocalization of SPARC, tenascin, and thrombospondin in pulmonary fibrosis.

C Kuhn 1, R J Mason 1
PMCID: PMC1869942  PMID: 7495300

Abstract

Several biochemically unrelated multifunctional extracellular proteins, SPARC, thrombospondin 1, and tenascin-C (TN), have been grouped as antiadhesive glycoproteins because they inhibit the spreading of cells on extracellular matrix in vitro. Migration of fibroblasts and epithelial cells into the air spaces to organize inflammatory exudate is a feature common to several fibrosing lung diseases. We hypothesized that migration would be facilitated by loosening the adhesive interactions between cells and the pericellular matrix components of the alveolar wall and that one or more of the anti-adhesive glycoproteins could be involved. Immunohistochemistry was used to localize SPARC, TN, and thrombospondin 1 in biopsies of organizing pneumonia, idiopathic pulmonary fibrosis (nine cases of usual interstitial pneumonia, one of desquamative interstitial pneumonia), and control lungs. Each antigen had a distinctive distribution. Only TN was expressed in control lungs, where it strongly stained the basement membrane of large bronchi and weakly stained alveolar entrance rings and small veins. In organizing pneumonia, TN was heavily stained through the entire extracellular matrix of the Masson bodies. In idiopathic pulmonary fibrosis, TN was abundant in the fibroblast foci of active fibrosis but was also present in the basement membrane regions beneath the metaplastic epithelium lining honeycomb cysts. TN was abundant in the interstitium in desquamative interstitial pneumonia. SPARC was observed only intracellularly where it occurred in the fibroblasts of Masson bodies of organizing pneumonia and the fibroblast foci of usual interstitial pneumonia. In desquamative interstitial pneumonia, expression of SPARC was minimal, in rare interstitial fibroblasts. Thrombospondin 1 was found consistently in organizing pneumonia but only infrequently in idiopathic pulmonary fibrosis. In both, it was localized in the extracellular matrix immediately beneath reparative epithelium. These results are consistent with a role for SPARC in fibroblast migration. TN may function in both fibroblast migration and the adhesion of metaplastic bronchial-type epithelium. However, these proteins also have other activities that may be important in pulmonary fibrosis. The localization of thrombospondin 1 suggests that it may be synthesized by regenerating epithelium where it may aid in the adhesion or migration of the epithelial cells.

Full text

PDF
1759

Images in this article

1761Figure 1
on p.1761
1761Figure 2
on p.1761
1762Figure 3
on p.1762
1763Figure 4
on p.1763
1764Figure 5
on p.1764
1765Figure 6
on p.1765
1766Figure 7
on p.1766

Selected References

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

  1. Anderson D. J., Blobel G. Immunoprecipitation of proteins from cell-free translations. Methods Enzymol. 1983;96:111–120. doi: 10.1016/s0076-6879(83)96012-3. [DOI] [PubMed] [Google Scholar]
  2. Antoniades H. N., Bravo M. A., Avila R. E., Galanopoulos T., Neville-Golden J., Maxwell M., Selman M. Platelet-derived growth factor in idiopathic pulmonary fibrosis. J Clin Invest. 1990 Oct;86(4):1055–1064. doi: 10.1172/JCI114808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Aufderheide E., Ekblom P. Tenascin during gut development: appearance in the mesenchyme, shift in molecular forms, and dependence on epithelial-mesenchymal interactions. J Cell Biol. 1988 Dec;107(6 Pt 1):2341–2349. doi: 10.1083/jcb.107.6.2341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Basset F., Ferrans V. J., Soler P., Takemura T., Fukuda Y., Crystal R. G. Intraluminal fibrosis in interstitial lung disorders. Am J Pathol. 1986 Mar;122(3):443–461. [PMC free article] [PubMed] [Google Scholar]
  5. Bonner J. C., Osornio-Vargas A. R., Badgett A., Brody A. R. Differential proliferation of rat lung fibroblasts induced by the platelet-derived growth factor-AA, -AB, and -BB isoforms secreted by rat alveolar macrophages. Am J Respir Cell Mol Biol. 1991 Dec;5(6):539–547. doi: 10.1165/ajrcmb/5.6.539. [DOI] [PubMed] [Google Scholar]
  6. Bornstein P. Thrombospondins: structure and regulation of expression. FASEB J. 1992 Nov;6(14):3290–3299. doi: 10.1096/fasebj.6.14.1426766. [DOI] [PubMed] [Google Scholar]
  7. Broekelmann T. J., Limper A. H., Colby T. V., McDonald J. A. Transforming growth factor beta 1 is present at sites of extracellular matrix gene expression in human pulmonary fibrosis. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6642–6646. doi: 10.1073/pnas.88.15.6642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chapman H. A., Allen C. L., Stone O. L. Abnormalities in pathways of alveolar fibrin turnover among patients with interstitial lung disease. Am Rev Respir Dis. 1986 Mar;133(3):437–443. doi: 10.1164/arrd.1986.133.3.437. [DOI] [PubMed] [Google Scholar]
  9. Chapman H. A., Jr, Bertozzi P., Reilly J. J., Jr Role of enzymes mediating thrombosis and thrombolysis in lung disease. Chest. 1988 Jun;93(6):1256–1263. doi: 10.1378/chest.93.6.1256. [DOI] [PubMed] [Google Scholar]
  10. Chiquet-Ehrismann R. Anti-adhesive molecules of the extracellular matrix. Curr Opin Cell Biol. 1991 Oct;3(5):800–804. doi: 10.1016/0955-0674(91)90053-2. [DOI] [PubMed] [Google Scholar]
  11. Chiquet-Ehrismann R., Kalla P., Pearson C. A., Beck K., Chiquet M. Tenascin interferes with fibronectin action. Cell. 1988 May 6;53(3):383–390. doi: 10.1016/0092-8674(88)90158-4. [DOI] [PubMed] [Google Scholar]
  12. Chiquet-Ehrismann R., Mackie E. J., Pearson C. A., Sakakura T. Tenascin: an extracellular matrix protein involved in tissue interactions during fetal development and oncogenesis. Cell. 1986 Oct 10;47(1):131–139. doi: 10.1016/0092-8674(86)90374-0. [DOI] [PubMed] [Google Scholar]
  13. End P., Panayotou G., Entwistle A., Waterfield M. D., Chiquet M. Tenascin: a modulator of cell growth. Eur J Biochem. 1992 Nov 1;209(3):1041–1051. doi: 10.1111/j.1432-1033.1992.tb17380.x. [DOI] [PubMed] [Google Scholar]
  14. Epler G. R., Colby T. V., McLoud T. C., Carrington C. B., Gaensler E. A. Bronchiolitis obliterans organizing pneumonia. N Engl J Med. 1985 Jan 17;312(3):152–158. doi: 10.1056/NEJM198501173120304. [DOI] [PubMed] [Google Scholar]
  15. Erickson H. P. Tenascin-C, tenascin-R and tenascin-X: a family of talented proteins in search of functions. Curr Opin Cell Biol. 1993 Oct;5(5):869–876. doi: 10.1016/0955-0674(93)90037-q. [DOI] [PubMed] [Google Scholar]
  16. Fukuda Y., Ishizaki M., Masuda Y., Kimura G., Kawanami O., Masugi Y. The role of intraalveolar fibrosis in the process of pulmonary structural remodeling in patients with diffuse alveolar damage. Am J Pathol. 1987 Jan;126(1):171–182. [PMC free article] [PubMed] [Google Scholar]
  17. Hasselaar P., Loskutoff D. J., Sawdey M., Sage E. H. SPARC induces the expression of type 1 plasminogen activator inhibitor in cultured bovine aortic endothelial cells. J Biol Chem. 1991 Jul 15;266(20):13178–13184. [PubMed] [Google Scholar]
  18. Hogg P. J., Stenflo J., Mosher D. F. Thrombospondin is a slow tight-binding inhibitor of plasmin. Biochemistry. 1992 Jan 14;31(1):265–269. doi: 10.1021/bi00116a036. [DOI] [PubMed] [Google Scholar]
  19. Hosokawa T., Muraishi A., Rothman V. L., Papale M., Tuszynski G. P. The effect of thrombospondin on invasion of fibrin gels by human A549 lung carcinoma. Oncol Res. 1993;5(4-5):183–189. [PubMed] [Google Scholar]
  20. Hølund B., Clausen P. P., Clemmensen I. The influence of fixation and tissue preparation on the immunohistochemical demonstration of fibronectin in human tissue. Histochemistry. 1981;72(2):291–299. doi: 10.1007/BF00517142. [DOI] [PubMed] [Google Scholar]
  21. Kelley J. Cytokines of the lung. Am Rev Respir Dis. 1990 Mar;141(3):765–788. doi: 10.1164/ajrccm/141.3.765. [DOI] [PubMed] [Google Scholar]
  22. Khalil N., O'Connor R. N., Unruh H. W., Warren P. W., Flanders K. C., Kemp A., Bereznay O. H., Greenberg A. H. Increased production and immunohistochemical localization of transforming growth factor-beta in idiopathic pulmonary fibrosis. Am J Respir Cell Mol Biol. 1991 Aug;5(2):155–162. doi: 10.1165/ajrcmb/5.2.155. [DOI] [PubMed] [Google Scholar]
  23. Kuhn C., 3rd, Boldt J., King T. E., Jr, Crouch E., Vartio T., McDonald J. A. An immunohistochemical study of architectural remodeling and connective tissue synthesis in pulmonary fibrosis. Am Rev Respir Dis. 1989 Dec;140(6):1693–1703. doi: 10.1164/ajrccm/140.6.1693. [DOI] [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. Lane T. F., Sage E. H. The biology of SPARC, a protein that modulates cell-matrix interactions. FASEB J. 1994 Feb;8(2):163–173. [PubMed] [Google Scholar]
  26. Lightner V. A., Erickson H. P. Binding of hexabrachion (tenascin) to the extracellular matrix and substratum and its effect on cell adhesion. J Cell Sci. 1990 Feb;95(Pt 2):263–277. doi: 10.1242/jcs.95.2.263. [DOI] [PubMed] [Google Scholar]
  27. Lotz M. M., Burdsal C. A., Erickson H. P., McClay D. R. Cell adhesion to fibronectin and tenascin: quantitative measurements of initial binding and subsequent strengthening response. J Cell Biol. 1989 Oct;109(4 Pt 1):1795–1805. doi: 10.1083/jcb.109.4.1795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mackie E. J., Halfter W., Liverani D. Induction of tenascin in healing wounds. J Cell Biol. 1988 Dec;107(6 Pt 2):2757–2767. doi: 10.1083/jcb.107.6.2757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mitchell D., Ibrahim S., Gusterson B. A. Improved immunohistochemical localization of tissue antigens using modified methacarn fixation. J Histochem Cytochem. 1985 May;33(5):491–495. doi: 10.1177/33.5.3921605. [DOI] [PubMed] [Google Scholar]
  30. Morla A., Zhang Z., Ruoslahti E. Superfibronectin is a functionally distinct form of fibronectin. Nature. 1994 Jan 13;367(6459):193–196. doi: 10.1038/367193a0. [DOI] [PubMed] [Google Scholar]
  31. Mornex J. F., Martinet Y., Yamauchi K., Bitterman P. B., Grotendorst G. R., Chytil-Weir A., Martin G. R., Crystal R. G. Spontaneous expression of the c-sis gene and release of a platelet-derived growth factorlike molecule by human alveolar macrophages. J Clin Invest. 1986 Jul;78(1):61–66. doi: 10.1172/JCI112574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Myers J. L., Katzenstein A. L. Epithelial necrosis and alveolar collapse in the pathogenesis of usual interstitial pneumonia. Chest. 1988 Dec;94(6):1309–1311. doi: 10.1378/chest.94.6.1309. [DOI] [PubMed] [Google Scholar]
  33. Palmer E. L., Rüegg C., Ferrando R., Pytela R., Sheppard D. Sequence and tissue distribution of the integrin alpha 9 subunit, a novel partner of beta 1 that is widely distributed in epithelia and muscle. J Cell Biol. 1993 Dec;123(5):1289–1297. doi: 10.1083/jcb.123.5.1289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Raines E. W., Lane T. F., Iruela-Arispe M. L., Ross R., Sage E. H. The extracellular glycoprotein SPARC interacts with platelet-derived growth factor (PDGF)-AB and -BB and inhibits the binding of PDGF to its receptors. Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1281–1285. doi: 10.1073/pnas.89.4.1281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Reed M. J., Puolakkainen P., Lane T. F., Dickerson D., Bornstein P., Sage E. H. Differential expression of SPARC and thrombospondin 1 in wound repair: immunolocalization and in situ hybridization. J Histochem Cytochem. 1993 Oct;41(10):1467–1477. doi: 10.1177/41.10.8245406. [DOI] [PubMed] [Google Scholar]
  36. Sage E. H., Bornstein P. Extracellular proteins that modulate cell-matrix interactions. SPARC, tenascin, and thrombospondin. J Biol Chem. 1991 Aug 15;266(23):14831–14834. [PubMed] [Google Scholar]
  37. Sage H., Farin F. M., Striker G. E., Fisher A. B. Granular pneumocytes in primary culture secrete several major components of the extracellular matrix. Biochemistry. 1983 Apr 26;22(9):2148–2155. doi: 10.1021/bi00278a015. [DOI] [PubMed] [Google Scholar]
  38. Sage H., Vernon R. B., Funk S. E., Everitt E. A., Angello J. SPARC, a secreted protein associated with cellular proliferation, inhibits cell spreading in vitro and exhibits Ca+2-dependent binding to the extracellular matrix. J Cell Biol. 1989 Jul;109(1):341–356. doi: 10.1083/jcb.109.1.341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Schultz-Cherry S., Murphy-Ullrich J. E. Thrombospondin causes activation of latent transforming growth factor-beta secreted by endothelial cells by a novel mechanism. J Cell Biol. 1993 Aug;122(4):923–932. doi: 10.1083/jcb.122.4.923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Silverstein R. L., Nachman R. L. Thrombospondin binds to monocytes-macrophages and mediates platelet-monocyte adhesion. J Clin Invest. 1987 Mar;79(3):867–874. doi: 10.1172/JCI112896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tremble P., Chiquet-Ehrismann R., Werb Z. The extracellular matrix ligands fibronectin and tenascin collaborate in regulating collagenase gene expression in fibroblasts. Mol Biol Cell. 1994 Apr;5(4):439–453. doi: 10.1091/mbc.5.4.439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Varani J., Nickoloff B. J., Riser B. L., Mitra R. S., O'Rourke K., Dixit V. M. Thrombospondin-induced adhesion of human keratinocytes. J Clin Invest. 1988 May;81(5):1537–1544. doi: 10.1172/JCI113486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Weinacker A., Ferrando R., Elliott M., Hogg J., Balmes J., Sheppard D. Distribution of integrins alpha v beta 6 and alpha 9 beta 1 and their known ligands, fibronectin and tenascin, in human airways. Am J Respir Cell Mol Biol. 1995 May;12(5):547–556. doi: 10.1165/ajrcmb.12.5.7537970. [DOI] [PubMed] [Google Scholar]
  45. Wight T. N., Raugi G. J., Mumby S. M., Bornstein P. Light microscopic immunolocation of thrombospondin in human tissues. J Histochem Cytochem. 1985 Apr;33(4):295–302. doi: 10.1177/33.4.3884704. [DOI] [PubMed] [Google Scholar]
  46. Wu P., Hoying J. B., Williams S. K., Kozikowski B. A., Lauffenburger D. A. Integrin-binding peptide in solution inhibits or enhances endothelial cell migration, predictably from cell adhesion. Ann Biomed Eng. 1994 Mar-Apr;22(2):144–152. doi: 10.1007/BF02390372. [DOI] [PubMed] [Google Scholar]
  47. Yokosaki Y., Palmer E. L., Prieto A. L., Crossin K. L., Bourdon M. A., Pytela R., Sheppard D. The integrin alpha 9 beta 1 mediates cell attachment to a non-RGD site in the third fibronectin type III repeat of tenascin. J Biol Chem. 1994 Oct 28;269(43):26691–26696. [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES