Skip to main content
SpringerLink
Log in

Locally applied GM-CSF induces the accumulation of α-smooth muscle act in containing myofibroblasts

  • Published:
Virchows Archiv B

Summary

We have examined the histological and cytoskeletal changes in rat connective tissues induced by subcutaneous perfusion with cytokines. Granulocyte macrophage-colony stimulating factor (GM-CSF), tumor necrosis factor-α (TNF-α), interleukin-1-α (IL-l-α), transforming growth factor-β (TGF-β) and platelet-derived growth factor (PDGF) produced a significant fibroblast accumulation, neovascular development and a weak to moderate leukocyte infiltration, while interleukin-2 (IL-2) and γ-interferon (γ-IFN) induced intense mononucleated leukocyte infiltration. Immunofluorescence staining showed that accumulated fibroblastic cells were positive for α-smooth muscle (SM) actin (but negative for the desmin and muscle myosin) only in GM-CSF-treated tissues. Electron microscopic examination established that a signifikant proportion of fibroblastic cell in GM-CSF-, IL-l-α- or TGF-β-treated animals were typical myofibroblasts. Only in GM-CSF-treated animals did microfilament bundles of myofibroblasts contain α-SM actin, when examined by immuno electron microscopy. Our results suggest that locally applied cytokines induce the formation of distinct granulation tissues. In particular, GM-CSF stimulates a-SM actin synthesis in myofibroblasts, illustrating an unexpected extra-hematopoietic in vivo effect of this factor.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Benzonana G, Skalli O, Gabbiani G (1988) Correlation between the distribution of smooth muscle or non muscle myosins and α-smooth muscle actin in normal and pathological soft tissues. Cell Mot Cytoskel 11:260–274

    Article  CAS  Google Scholar 

  • Bussolino F, Wang JM, Defilippi P, Turrini F, Sanavio F, Edgell CJS, Aglietta M, Arese P, Mantovani A (1989) Granulocyte- and granulocyte-macrophage-colony stimulating factors induce human endothelial cells to migrate and proliferate. Nature 337:471–473

    Article  PubMed  CAS  Google Scholar 

  • Clark SC, Kamen R (1987) The human hematopoietic colony-stimulating factors. Science 236:1229–1237

    Article  PubMed  CAS  Google Scholar 

  • Czaja MJ, Weiner FR, Flanders KC, Gimbrone MA, Wind R, Biempica L, Zern MA (1989) In vitro and in vivo association of transforming growth factor-β-1 with hepatic fibrosis. J Cell Biol 108:2477–2482

    Article  PubMed  CAS  Google Scholar 

  • Danscher G (1981) Localization of gold in biological tissue. A photochemical method for light and electronmicroscopy. Histochemistry 71:81–88

    Article  PubMed  CAS  Google Scholar 

  • Darby I, Skalli O, Gabbiani G (1990) α-Smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab Invest 63:21–29

    PubMed  CAS  Google Scholar 

  • Dedhar S, Gaboury L, Galloway P, Eaves C (1988) Human granulocyte-macrophage colony-stimulating factor is a growth factor active on a variety of cell types of nonhemopoietic origin. Proc Natl Acad Sci USA 85:9253–9257

    Article  PubMed  CAS  Google Scholar 

  • DeLamarter JF, Mermod JJ, Liang CM, Eliason JF, Thatcher DR (1985) Recombinant murine GM-CSF from E. coli has biological activity and is neutralized by a specific antiserum. EMBO J 4:2575–2581

    Google Scholar 

  • Duncan MR, Berman B (1985) γ Interferon is the lymphokine and β interferon the monokine responsible for inhibition of fibroblast collagen production and late but not early fibroblast proliferation. J Exp Med 162:516–527

    Article  PubMed  CAS  Google Scholar 

  • Freundlich B, Bomalaski JS, Neilson E, Jimenez SA (1986) Regulation of fibroblast proliferation and collagen synthesis by cytokines. Immunol Today 7:303–307

    Article  CAS  Google Scholar 

  • Gabbiani G, Majno G (1972) Dupuytren’s contracture: fibroblast contraction? An ultrastructural study. Am J Pathol 66:131–146

    PubMed  CAS  Google Scholar 

  • Gabbiani G, Ryan GB, Majno G (1971) Presence of modified fibroblasts in granulation tissue and their possible role in wound contraction. Experienta 27:549–550

    Article  CAS  Google Scholar 

  • Giri S, Hyde DM (1988) Ameliorating effect of an interferon inducer polyinosinic-polycytidylic acid on bleomycin-induced lung fibrosis in hamsters. Am J Pathol 133:525–536

    PubMed  CAS  Google Scholar 

  • Greenhalgh DG, Sprugel KH, Murray MJ, Ross R (1990) PDGF and FGF stimulate wound healing in the genetically diabetic mouse. Am J Pathol 136:1235–1246

    PubMed  CAS  Google Scholar 

  • Hansson GK, Hellstrand M, Rymo L, Rubbia L, Gabbiani G (1989) Interferon γ inhibits both proliferation and expression of differentiation-specific α-smooth muscle actin in arterial smooth muscle cells. J Exp Med 170:1595–1608

    Article  PubMed  CAS  Google Scholar 

  • Hasegawa T, Hirose T, Kudo E, Abe JI, Hizawa K (1990) Cytoskeletal characteristics of myofibroblasts in benign neoplastic and reactive fibroblastic lesions. Virchows Arch [A] 416:375–382

    CAS  Google Scholar 

  • Hyde DM, Henderson TS, Giri SN, Tyler NK, Stovall MY (1988) Effect of murine gamma interferon on cellular responses to bleomycin in mice. Exp Lung Res 14:687–704

    Article  PubMed  CAS  Google Scholar 

  • Jimenez SA, Freundlich B, Rosenbloom J (1984) Selective inhibition of human diploid fibroblast collagen synthesis by interferons. J Clin Invest 74:1112–1116

    Article  PubMed  CAS  Google Scholar 

  • Kapanci Y, Burgan S, Pietra GG, Conne B, Gabbiani G (1990) Modulation of actin isoform expression in alveolar myofibroblasts (contractile interstitial cells) during pulmonary hypertension. Am J Pathol 136:881–889

    PubMed  CAS  Google Scholar 

  • Kocher O, Skalli O, Bloom WS, Gabbiani G (1984) Cytoskeleton of rat aortic smooth muscle cells. Normal conditions and experimental intimai thickening. Lab Invest 50:645–652

    PubMed  CAS  Google Scholar 

  • Lang RA, Metcalf D, Cuthbertson RA, Lyons I, Stanley E, Kelso A, Kannourakis G, Williamson DJ, Klintworth GK, Gonda TJ, Dunn AR (1987) Transgenic mice expressing a hemopoietic growth factor gene (GM-CSF) develop accumulations of macrophages, blindness, and a fatal syndrome of tissue damage. Cell 51:675–686

    Article  PubMed  CAS  Google Scholar 

  • Le J, Vilcek J (1987) Tumor necrosis factor and interleukin 1: cytokines with multiple overlapping biological activities. Lab Invest 56:234–248

    PubMed  CAS  Google Scholar 

  • Lennette DA (1978) An improved mounting medium for immunofluorescent micrscopy. Am J Clin Pathol 69:647–648

    PubMed  CAS  Google Scholar 

  • Lomedico PT, Gubler U, Hellmann CP, Dukovich M, Giri JG, Pan YCE, Collier K, Semionow R, Chua AO, Mizel SB (1984) Cloning and expression of murine interleukin-1 cDNA in Escherichia coli. Nature 312:458–462

    Article  PubMed  CAS  Google Scholar 

  • MacDonald D, Adams JA, McCarthy D, Barrett AJ (1990) Interleukin-2 inhibits growth of fibroblasts derived from human bone marrow. Acta Haemat 83:26–30

    PubMed  CAS  Google Scholar 

  • Majno G, Gabbiani G, Hirschel BJ, Ryan GB, Statkov PR (1971) Contraction of granulation tissue in vitro: similarity to smooth muscle. Science 173:548–550

    Article  PubMed  CAS  Google Scholar 

  • Mantovani A, Dejana E (1989) Cytokines as communication signals between leukocytes and endothelial cells. Immunol Today 10:370–375

    Article  PubMed  CAS  Google Scholar 

  • Martinet Y, Rom WN, Grotendorst GR, Martin GR, Crystal RG (1987) Exaggerated spontaneous release of platelet-derived growth factor by alveolar macrophages from patients with idiopathic pulmonary fibrosis. N Engl J Med 317:202–209

    PubMed  CAS  Google Scholar 

  • Mitchell J, Woodcock-Mitchell J, Reynolds S, Low R, Leslie K, Adler K, Gabbiani G, Skalli O (1989) σ-Smooth muscle actin in parenchymal cells of bleomycin-injured rat lung. Lab Invest 60:643–650

    PubMed  CAS  Google Scholar 

  • Mustoe TA, Pierce GF, Thomason A, Gramates P, Sporn MB, Deuel TF (1987) Accelerated healing of incisional wounds in rats induced by transforming growth factor-β. Science 237:1333–1336

    Article  PubMed  CAS  Google Scholar 

  • Piguet PF, Collart MA, Grau GE, Kapanci Y, Vassali P (1989) Tumor necrosis factor/cachectin plays a key role in bleomycininduced pneumopathy and fibrosis. J Exp Med 170:655–663

    Article  PubMed  CAS  Google Scholar 

  • Piquet PF, Grau GE, Vassali P (1990) Subcutaneous perfusion of tumor necrosis factor induces local proliferation of fibroblasts, capillaries, and epidermal cells, or massive tissue necrosis. Am J Pathol 136:103–110

    Google Scholar 

  • Pojda Z, Molineux G, Dexter TM (1989) Effects of long-term in vivo treatment of mice with purified murine recombinant GMCSF. Exp Hematol 17:1100–1104

    PubMed  CAS  Google Scholar 

  • Roberts AB, Sporn MB, Assoian RK, Smith JM, Roche NS, Wakefield LM, Heine UI, Liotta LA, Falanga V, Kehrl JH, Fauci AS (1986) Transforming growth factor type β: rapid induction of fibrosis and angiogenesis in vivo and stimulation of collagen formation in vitro. Proc Natl Acad Sci USA 83:4167–4171

    Article  PubMed  CAS  Google Scholar 

  • Ross R, Raines EW, Bowen-Pope DF (1986) The biology of platelet-derived growth factor. Cell 46:155–169

    Article  PubMed  CAS  Google Scholar 

  • Rubbia L, Sappino AP, Hansson GK, Gabbiani G (1989) Action of different cytokines on actin isoform expression on fibroblasts in vitro. Experientia 45: A49

    Google Scholar 

  • Sappino AP, Skalli O, Jackson B, Schürch W, Gabbiani G (1988) Smooth-muscle differentiation in stromal cells of malignant and non-malignant breast tissues. Int J Cancer 41:707–712

    Article  PubMed  CAS  Google Scholar 

  • Sappino AP, Dietrich PY, Skalli O, Widgren S, Gabbiani G (1989) Colonic pericryptal fibroblasts. Differentiation pattern in embryogenesis and phenotypic modulation in epithelial proliferative lesions. Virchows Arch [A] 415:551–557

    CAS  Google Scholar 

  • Sappino AP, Schürch W, Gabbiani G (1990) The differentiation repertoire of fibroblastic cells: expression of cytoskeletal proteins as marker of phenotypic modulations. Lab Invest 63:144–161

    PubMed  CAS  Google Scholar 

  • Seemayer TA, Lagacé R, Schürch W, Tremblay G (1979) Myofibroblasts in the stroma of invasive and metastatic carcinoma: a possible host response to neoplasia. Am J Surg Pathol 3:525–533

    Article  PubMed  CAS  Google Scholar 

  • Skalli O, Gabbiani G (1988) The biology of the myofibroblast. Relationship to wound contraction and fibrocontractive diseases. In: Clark RAF, Henson PM (eds) The molecular and cellular biology of wound repair. Plenum Publishing Corporation, New York, pp 373–402

    Google Scholar 

  • Skalli O, Ropraz P, Trzeciak A, Benzonana G, Gillessen D, Gabbiani G (1986) A monoclonal antibody against α-smooth muscle actin: a new probe for smooth muscle differentiation. J Cell Biol 103:2787–2796

    Article  PubMed  CAS  Google Scholar 

  • Skalli O, Vandekerckhove J, Gabbiani G (1987) Actin-isoform pattern as a marker of normal or pathological smooth-muscle and fibroblastic tissues. Differentiation 33:232–238

    Article  PubMed  CAS  Google Scholar 

  • Skalli O, Schürch W, Seemayer T, Lagacé R, Montandon D, Pittet B, Gabbiani G (1989 a) Myofibroblasts from diverse pathologic settings are heterogeneous in their content of actin isoforms and intermediate filament proteins. Lab Invest 60:275–285

    PubMed  CAS  Google Scholar 

  • Skalli O, Pelte MF, Peclet MC, Gabbiani G, Gugliotta P, Bussolati G, Ravazzola M, Orci L (1989b) α-Smooth muscle actin, a differentiation marker of smooth muscle cells, is present in microfilamentous bundles of pericytes. J Histochem Cytochem 37:315–321

    PubMed  CAS  Google Scholar 

  • Sporn MB, Roberts AB (1986) Peptide growth factors and inflammation, tissue repair, and cancer. J Clin Invest 78:329–332

    Article  PubMed  CAS  Google Scholar 

  • Sprugel KH, McPherson JM, Clowes AW, Ross R (1987) Effects of growth factors in vivo, I. Cell ingrowth into porous subcutaneous chambers. Am J Pathol 129:601–613

    PubMed  CAS  Google Scholar 

  • Trincherini G, Perussia B (1985) Immune interferon: a pleiotropic lymphokine with multiple effects. Immunol Today 6:131–136

    Article  Google Scholar 

  • Van der Meide PH, Dubbled M, Vijverberg K, Kos T, Schellekins H (1986) The purification and characterization of rat gamma interferon by use of two monoclonal antibodies. J Gen Virol 67:1059–1065

    Article  PubMed  Google Scholar 

  • Wahl SM, McCartney-Francis N, Mergenhagen SE (1989) Inflammatory and immunomodulatory roles of TGF-β. Immunol Today 8:258–261

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pathology, University of Geneva, CMU, 1 rue Michel-Servet, CH-1211, Geneva 4, Switzerland

    Laura Rubbia-Brandt & Giulio Gabbiani

  2. Division of Onco-Hematology, University Hospital of Geneva, CH-1211, Geneva 4, Switzerland

    André-Pascal Sappino

Authors
  1. Laura Rubbia-Brandt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. André-Pascal Sappino
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Giulio Gabbiani
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rubbia-Brandt, L., Sappino, AP. & Gabbiani, G. Locally applied GM-CSF induces the accumulation of α-smooth muscle act in containing myofibroblasts. Virchows Archiv B Cell Pathol 60, 73–82 (1991). https://doi.org/10.1007/BF02899530

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02899530

Key words

Search

Navigation