Cell and Tissue Research

, Volume 281, Issue 2, pp 231–242 | Cite as

The monoclonal antibody GB 42 — a useful marker for the differentiation of myofibroblasts

  • Gaby Kohnen
  • Mario Castellucci
  • Bae-Li Hsi
  • Chang-Jing G. Yeh
  • Peter Kaufmann


The expression patterns of a variety of cytoskeletal antigens were studied in normal human tissues (placenta, umbilical cord, myometrium, colon, mammary gland, testis, skeletal muscle, myocardium) as well as in abnormal human tissues (palmar fibromatosis, fibrocystic disease of the mammary gland, mammary carcinoma). The immunohistochemical binding patterns of the monoclonal antibody GB 42 were compared to those of commerical antibodies directed against vimentin, desmin, smooth muscle myosin, pan actin, α-smooth muscle actin and γ-smooth muscle actin. Methods applied comprised immunohistochemistry on cryostat sections and paraffin sections. Immunogold immunocytochemistry was performed on Lowicryl sections. The patterns of GB 42-binding were confirmed biochemically by SDS-PAGE and Western-blotting, and quantitative amino acid analysis. Our data suggest that the monoclonal antibody GB 42 recognizes an actin isoform which is identical to, or closely related to, γ-smooth muscle actin. Unlike the commercially available antibody against γ-smooth muscle actin, GB 42 does not cross-react with α-skeletal or α-cardiac actins. The GB 42-antigen is expressed in smooth muscle cells, myoepithelial cells and in later stages of differentiation of myofibroblasts, in all the tissues investigated. Throughout the development of smooth muscle cells and myofibroblasts, the appearance of the GB 42-antigen occurs after the expression of vimentin, desmin and α-smooth muscle actin, but prior to the expression of smooth muscle myosin. GB 42 is a reliable marker for higher stages of differentiation of smooth muscle cells and myofibroblasts.

Key words

Placenta Stem villi Actin isoforms Myofibroblasts Smooth muscle cells Immunohisto-chemistry Human 


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  1. Barja F, Coughlin C, Belin D, Gabbiani G (1986) Actin isoform synthesis and mRNA levels in quiescent and proliferating rat aortic smooth muscle cells in vivo and in vitro. Lab Invest 55:226–233Google Scholar
  2. Benirschke K, Kaufmann P (1994) Pathology of the Human Placenta, 3rd edn. Springer, New YorkGoogle Scholar
  3. Benyamin Y, Roustan C, Boyer M (1986) Anti-actin antibodies. Chemical modification allows the selective production of antibodies to the N-terminal region. J Immunol Meth 86:21–29Google Scholar
  4. 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 Motil Cytoskeleton 11:260–274Google Scholar
  5. Bergsma D, Chang K, Schwartz R (1985) Novel chicken actin gene: Third cytoplasmic isoform. Mol Cell Biol 5:1151–1162Google Scholar
  6. Bulinski JC, Kumar S, Titani K, Hauschka SD (1983) Peptide antibody specific for the amino-terminus of skeletal muscle α-actin. Proc Natl Acad Sci USA 80:1506–1510Google Scholar
  7. Chamley JH, Gröschel-Stewart U, Campbell GR, Burnstock G (1977) Distinction between smooth muscle, fibroblasts and endothelial cells in culture by use of fluoresceinated antibodies against smooth muscle actin. Cell Tissue Res 177:445–457Google Scholar
  8. Danscher G, Norgaard JOR (1983) Light microscopic visualization of colloidal gold on resin embedded tissue. J Histochem Cytochem 31:1394–1398Google Scholar
  9. Darby I, Skalli O, Gabbiani G (1990) α-Smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab Invest 63:21–29Google Scholar
  10. Davidoff MS, Breucker H, Holstein AF, Seild K (1990) Cellular architecture of the lamina propria of human seminiferous tubules. Cell Tissue Res 262:253–261Google Scholar
  11. Debus E, Weber K, Osborn M (1983) Monoclonal antibodies to desmin, the muscle specific intermediate filament protein. EMBO 2:2305–2312Google Scholar
  12. Demir R, Demir N, Kohnen G, Kosanke G, Mironov AV, Üstünel I, Kocamaz E (1992) Ultrastructure and distribution of myofibroblast-like cells in human placental stem villi. Electron Microscopy 3:509–510Google Scholar
  13. Fyrberg EA, Mahaffey JW, Bond BJ, Davidson N (1983) Transcripts of the sixDrosophila actin genes accumulate in a stage-specific and tissue-specific manner. Cell 33:115–123Google Scholar
  14. Gabbiani G, Maino G (1972) Dupuytren's contracture: Fibroblast contraction. Am J Pathol 66:131–146Google Scholar
  15. Gabbiani G, Hirschel BJ, Ryan GB, Statkov PR, Majno G (1972) Granulation tissue as a contractile organ. J Exper Med 135: 719–734Google Scholar
  16. Gabbiani G, Schmid E, Winter S, Chaponnier C, Chastonay C de, Vandekerckhove J, Weber K, Franke WW (1981) Vascular smooth muscle cells differ from other smooth muscle cells: Predominance of vimentin filaments and a specific α-type actin. Proc Natl Acad Sci USA 78:298–302Google Scholar
  17. Garcia R, Paz-Aliaga B, Ernst SG, Crain WR (1984) Three sea urchin actin genes show different patterns of expression: muscle specific, embryo-specific, and constitutive. Mol Cell Biol 4: 840–845Google Scholar
  18. Giloh H, Sedat JW (1982) Fluorescence microscopy: reduced photobleaching of rhodamine and fluorescein protein conjugates by n-propyl-gallate. Science 217:1252–1255Google Scholar
  19. Graf R, Frank H-G, Öney T (1992) Histochemical and immunocytochemical investigations of the fetal extravascular and vascular contractile system in the normal placenta and during preeclampsia. In: Neubert D (ed) Risk Assessment of Prenatally-Induced Adverse Health Effects. Springer, Berlin Heidelberg New York, pp 537–550Google Scholar
  20. Güldner FH, Wolff JR, Graf v. Deyserlink D (1972) Fibroblasts as a part of the contractile system in duodenal villi of rat. Z Zellforsch 135:349–360Google Scholar
  21. Herman IM, D'Amore PA (1985) Microvascular pericytes contain muscle and non-muscle actins. J Cell Biol 101:43–52Google Scholar
  22. Hsi B-L, Yeh C-JG (1987) Studies of C1q deposits in the human placenta using monoclonal antibodies to human extra-embryonic tissues. Trophoblast Res 2:223–231Google Scholar
  23. Hsi B-L, Yeh C-JG (1988) Monoclonal antibodies to placental vascular structures. Trophoblast Res 3:139–148Google Scholar
  24. 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 136:881–889Google Scholar
  25. Kocher O, Skalli O, Bloom WS, Gabbiani G (1984) Cytoskeleton of rat aortic smooth muscle cells. Lab Invest 50:645–652Google Scholar
  26. Kocher O, Skalli O, Cerutti D, Gabbiani F, Gabbiani G (1985) Cytoskeletal features of rat aortic cells during development. Circ Res 56:829–838Google Scholar
  27. Kocher O, Gabbiani F, Gabbiani G, Reidy MA, Sami Cokay M, Peters H, Hüttner I (1991) Phenotypic features of smooth muscle cells during the evolution of experimental carotid artery intimal thickening. Lab Inv 65:459–470Google Scholar
  28. Kohnen O, Mironov V, Demir R, Castellucci M, Kaufmann P (1992) Immunhistochemische Klassifizierung von Stammzotten in der menschlichen Placenta. Anat Anz 174 [Suppl]:127Google Scholar
  29. Kohnen G, Castellucci M, Graf R, Kaufmann P (1993a) Contractile filaments of extravascular stromal cells in human placental stem villi (abstract): Placenta 14:A38Google Scholar
  30. Kohnen G, Mironov V, Demir R, Castellucci M, Kaufmann P (1993b) Immunhistochemie und Ultrastruktur kontraktiler Zellen im Zottenstroma der menschlichen Placenta. Annals Anat 175 [Suppl]:37–38Google Scholar
  31. Kohnen G, Kaufmann P (1994) Expressionsmuster cytoskeletaler Proteine in der Entwicklung des Zottenstromas der menschlichen Placenta. Annals Anat 176 [Suppl]:10Google Scholar
  32. Kuhn C, McDonald JA (1991) The roles of the myofibroblast in idiopathic pulmonary fibrosis. Am J Pathol 138:1257–1265Google Scholar
  33. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685Google Scholar
  34. Leslie KO, Mitchell J, Low R (1992) Views and Reviews. Lung Myofibroblasts. Cell Mot Cytoskeleton 22:92–98Google Scholar
  35. Lessard JL (1988) Two monoclonal antibodies to actin: one muscle selective and one generally reactive. Cell Mot Cytoskeleton 10:349–362Google Scholar
  36. Lessard JL, Scheffter S, Engel L, Teppermann K (1983) Immunofluorescent localization of actins in differentiating chick myoblasts (abstract). J Cell Biol 97 (5 Pt 2):74aGoogle Scholar
  37. Longtine JA, Pinkus GS, Fujiwara K, Corson JM (1985) Immunohistochemical localization of smooth muscle myosin in normal human tissues. J Histochem Cytochem 33:179–184Google Scholar
  38. Lowry OH, Rosebrough NJ, Farr AL, Randal RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  39. Majno G, Gabbiani G, Hirschel BJ, Ryan GB, Statkov PR (1971) Contraction of granulation tissue in vitro: Similarity to smooth muscle cells. Science 173:548–550Google Scholar
  40. McHugh KM, Lessard JL (1988) The development expressio of the rat α-vascular and γ-enteric smooth muscle isoactins: Isolation and characterization of a rat γ-enteric actin cDNA. Mol Cell Biol 8:5224–5231Google Scholar
  41. Mohun TJ, Brennan S, Dathan N, Fairman S, Gordon JB (1984) Cell type-specific activation of actin genes in the early amphibian embryo Nature 311:716–721Google Scholar
  42. Nanaev AK, Shirinsky VP, Birukov KG (1991) Immunofluorescent study of heterogeneity in smooth muscle cells of human fetal vessels using antibodies to myosin, desmin, and vimentin. Cell Tissue Res 266:535–540Google Scholar
  43. Osborn M, Debus E, Weber K (1984) Monoclonal antibodies specific for vimentin. Eur J Cell Biol 34:137–143Google Scholar
  44. Otey CA, Kalnoski MH, Lessard JL, Bulinski JC (1986) Immunolocalization of the gamma isoform of nonmuscle actin in cultured cells. J Cell Biol 102:1726–1737Google Scholar
  45. Owens GK, Loeb A, Gordon D, Thompson MM (1986) Expression of smooth muscle-specific α-isoactin in cultured vascular smooth muscle cells: relationship between growth and cytodifferentiation. J Cell Biol 102:343–352Google Scholar
  46. Sadano H, Taniguchi S, Baba T (1990) Newly identified type of β-actin reduces invasiveness of mouse B 16-melanoma. FEBS Lett 271:23–27Google Scholar
  47. Sappino A-P, 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 41:707–712Google Scholar
  48. Sappino AP, Schürch W, Gabbiani G (1990) Biology of disease. Differentiation repertoire of fibroblastic cells: Expression of cytoskeletal proteins as marker of phenotypic modulations. Lab Invest 63:144–161Google Scholar
  49. Sawtell N, hartman A, Lessard JL (1988) Unique isoactins in the brush border of rat intestinal epithelial cells. Cell Mot Cytoskeleton 11:318–325Google Scholar
  50. Schürch W, Seemayer TA, Lagace R (1981) Stromal myofibroblasts in primary invasive and metastic carcinomas. Virchows Arch [A] 391:125–139Google Scholar
  51. Seemayer TA, Lagace R, Schürch W, Thelmo W (1980) The myofibroblast: Biologic, pathologic and theoretical considerations. Pathol Annual 15:443–470Google Scholar
  52. Skalli O, Ropraz P, Trecziak A, Benzonana G, Gilessen D, Gabbiani G (1986) A monoclonal antibody against α-smooth muscle actin: a new probe for smooth muscle differentiation. J Cell Biol 103:2787–2796Google Scholar
  53. Skalli O, Vandekerckhove J, Gabbiani G (1987) Actin-isoform pattern as a marker of normal or pathological smooth-muscle and fibroblastic tissues. Differentiation 33:232–238Google Scholar
  54. Skalli O, Schürch W, Seemayer T, Lagace R, Montandon D, Pittet B, Gabbiani G (1989) Myofibroblasts from diverse pathologic settings are heterogeneous in their content of actin isoforms and intermediate filament proteins. Lab Invest 60:275–285Google Scholar
  55. Spackman DH, Stein WH, Moore S (1958) Automatic recording apparatus for use in the chromatography of amino acids. Anal Chem 30:1190–1206Google Scholar
  56. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354Google Scholar
  57. Vandekerckhove J, Weber K (1978) At least six different actins are expressed in a higher mammal: An analysis based on the amino acid sequence of the amino-terminal tryptic peptide. J Mol Biol 126:783–802Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Gaby Kohnen
    • 1
  • Mario Castellucci
    • 1
  • Bae-Li Hsi
    • 2
  • Chang-Jing G. Yeh
    • 2
  • Peter Kaufmann
    • 1
  1. 1.Department of AnatomyTechnical University of AachenAachenGermany
  2. 2.INSERM U 354NiceFrance

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