[go: up one dir, main page]
More Web Proxy on the site http://driver.im/ Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1993 May 2;121(4):931–943. doi: 10.1083/jcb.121.4.931

Regulation of in vitro capillary tube formation by anti-integrin antibodies

PMCID: PMC2119791  PMID: 8491784

Abstract

Human endothelial cells are induced to form an anastomosing network of capillary tubes on a gel of collagen I in the presence of PMA. We show here that the addition of mAbs, AK7, or RMAC11 directed to the alpha chain of the major collagen receptor on endothelial cells, the integrin alpha 2 beta 1, enhance the number, length, and width of capillary tubes formed by endothelial cells derived from umbilical vein or neonatal foreskins. The anti-alpha 2 beta 1 antibodies maintained the endothelial cells in a rounded morphology and inhibited both their attachment to and proliferation on collagen but not on fibronectin, laminin, or gelatin matrices. Furthermore, RMAC11 promoted tube formation in collagen gels of increased density which in the absence of RMAC11 did not allow tube formation. Neither RMAC11 or AK7 enhanced capillary formation in the absence of PMA. Lumen structure and size were also altered by antibody RMAC11. In the absence of antibody the majority of lumina were formed intracellularly from single cells, but in the presence of RMAC11, multiple cells were involved and the lumen size was correspondingly increased. Endothelial cells were also induced to undergo capillary formation in fibrin gels after PMA stimulation. The addition of anti-alpha v beta 3 antibodies promoted tube formation in fibrin gels and inhibited EC adhesion to and proliferation on a fibrinogen matrix. The enhancement of capillary formation by the anti- integrin antibodies was matrix specific; that is, anti-alpha v beta 3 antibodies only enhanced tube formation on fibrin gels and not on collagen gels while anti-alpha v beta 1 antibodies only enhanced tubes on collagen and not on fibrin gels. Thus we postulate that changes in the adhesive nature of endothelial cells for their extracellular matrix can profoundly effect their function. Anti-integrin antibodies which inhibit cell-matrix interactions convert endothelial cells from a proliferative phenotype towards differentiation which results in enhanced capillary tube formation.

Full Text

The Full Text of this article is available as a PDF (4.6 MB).

Selected References

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

  1. Aggeler J., Frisch S. M., Werb Z. Changes in cell shape correlate with collagenase gene expression in rabbit synovial fibroblasts. J Cell Biol. 1984 May;98(5):1662–1671. doi: 10.1083/jcb.98.5.1662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albelda S. M., Buck C. A. Integrins and other cell adhesion molecules. FASEB J. 1990 Aug;4(11):2868–2880. [PubMed] [Google Scholar]
  3. Albelda S. M., Daise M., Levine E. M., Buck C. A. Identification and characterization of cell-substratum adhesion receptors on cultured human endothelial cells. J Clin Invest. 1989 Jun;83(6):1992–2002. doi: 10.1172/JCI114109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ben-Ze'ev A., Farmer S. R., Penman S. Protein synthesis requires cell-surface contact while nuclear events respond to cell shape in anchorage-dependent fibroblasts. Cell. 1980 Sep;21(2):365–372. doi: 10.1016/0092-8674(80)90473-0. [DOI] [PubMed] [Google Scholar]
  5. Ben-Ze'ev A., Robinson G. S., Bucher N. L., Farmer S. R. Cell-cell and cell-matrix interactions differentially regulate the expression of hepatic and cytoskeletal genes in primary cultures of rat hepatocytes. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2161–2165. doi: 10.1073/pnas.85.7.2161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carter W. G., Wayner E. A., Bouchard T. S., Kaur P. The role of integrins alpha 2 beta 1 and alpha 3 beta 1 in cell-cell and cell-substrate adhesion of human epidermal cells. J Cell Biol. 1990 Apr;110(4):1387–1404. doi: 10.1083/jcb.110.4.1387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chan B. M., Kassner P. D., Schiro J. A., Byers H. R., Kupper T. S., Hemler M. E. Distinct cellular functions mediated by different VLA integrin alpha subunit cytoplasmic domains. Cell. 1992 Mar 20;68(6):1051–1060. doi: 10.1016/0092-8674(92)90077-p. [DOI] [PubMed] [Google Scholar]
  8. Cheresh D. A., Spiro R. C. Biosynthetic and functional properties of an Arg-Gly-Asp-directed receptor involved in human melanoma cell attachment to vitronectin, fibrinogen, and von Willebrand factor. J Biol Chem. 1987 Dec 25;262(36):17703–17711. [PubMed] [Google Scholar]
  9. Defilippi P., Truffa G., Stefanuto G., Altruda F., Silengo L., Tarone G. Tumor necrosis factor alpha and interferon gamma modulate the expression of the vitronectin receptor (integrin beta 3) in human endothelial cells. J Biol Chem. 1991 Apr 25;266(12):7638–7645. [PubMed] [Google Scholar]
  10. Defilippi P., van Hinsbergh V., Bertolotto A., Rossino P., Silengo L., Tarone G. Differential distribution and modulation of expression of alpha 1/beta 1 integrin on human endothelial cells. J Cell Biol. 1991 Aug;114(4):855–863. doi: 10.1083/jcb.114.4.855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. DiPersio C. M., Jackson D. A., Zaret K. S. The extracellular matrix coordinately modulates liver transcription factors and hepatocyte morphology. Mol Cell Biol. 1991 Sep;11(9):4405–4414. doi: 10.1128/mcb.11.9.4405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Doctrow S. R., Folkman J. Protein kinase C activators suppress stimulation of capillary endothelial cell growth by angiogenic endothelial mitogens. J Cell Biol. 1987 Mar;104(3):679–687. doi: 10.1083/jcb.104.3.679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Duband J. L., Dufour S., Yamada K. M., Thiery J. P. The migratory behavior of avian embryonic cells does not require phosphorylation of the fibronectin-receptor complex. FEBS Lett. 1988 Mar 28;230(1-2):181–185. doi: 10.1016/0014-5793(88)80667-7. [DOI] [PubMed] [Google Scholar]
  14. Elsdale T., Bard J. Collagen substrata for studies on cell behavior. J Cell Biol. 1972 Sep;54(3):626–637. doi: 10.1083/jcb.54.3.626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Folkman J. Angiogenesis: initiation and control. Ann N Y Acad Sci. 1982;401:212–227. doi: 10.1111/j.1749-6632.1982.tb25720.x. [DOI] [PubMed] [Google Scholar]
  16. Folkman J., Haudenschild C. Angiogenesis in vitro. Nature. 1980 Dec 11;288(5791):551–556. doi: 10.1038/288551a0. [DOI] [PubMed] [Google Scholar]
  17. Folkman J. How is blood vessel growth regulated in normal and neoplastic tissue? G.H.A. Clowes memorial Award lecture. Cancer Res. 1986 Feb;46(2):467–473. [PubMed] [Google Scholar]
  18. Folkman J., Klagsbrun M. Angiogenic factors. Science. 1987 Jan 23;235(4787):442–447. doi: 10.1126/science.2432664. [DOI] [PubMed] [Google Scholar]
  19. Folkman J., Moscona A. Role of cell shape in growth control. Nature. 1978 Jun 1;273(5661):345–349. doi: 10.1038/273345a0. [DOI] [PubMed] [Google Scholar]
  20. Glowacki J., Trepman E., Folkman J. Cell shape and phenotypic expression in chondrocytes. Proc Soc Exp Biol Med. 1983 Jan;172(1):93–98. doi: 10.3181/00379727-172-41533. [DOI] [PubMed] [Google Scholar]
  21. Gospodarowicz D., Greenburg G., Birdwell C. R. Determination of cellular shape by the extracellular matrix and its correlation with the control of cellular growth. Cancer Res. 1978 Nov;38(11 Pt 2):4155–4171. [PubMed] [Google Scholar]
  22. Greenburg G., Hay E. D. Epithelia suspended in collagen gels can lose polarity and express characteristics of migrating mesenchymal cells. J Cell Biol. 1982 Oct;95(1):333–339. doi: 10.1083/jcb.95.1.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hemler M. E., Elices M. J., Chan B. M., Zetter B., Matsuura N., Takada Y. Multiple ligand binding functions for VLA-2 (alpha 2 beta 1) and VLA-3 (alpha 3 beta 1) in the integrin family. Cell Differ Dev. 1990 Dec 2;32(3):229–238. doi: 10.1016/0922-3371(90)90035-u. [DOI] [PubMed] [Google Scholar]
  24. Hynes R. O. Integrins: a family of cell surface receptors. Cell. 1987 Feb 27;48(4):549–554. doi: 10.1016/0092-8674(87)90233-9. [DOI] [PubMed] [Google Scholar]
  25. Ingber D. E., Folkman J. How does extracellular matrix control capillary morphogenesis? Cell. 1989 Sep 8;58(5):803–805. doi: 10.1016/0092-8674(89)90928-8. [DOI] [PubMed] [Google Scholar]
  26. Ingber D. E., Folkman J. Mechanochemical switching between growth and differentiation during fibroblast growth factor-stimulated angiogenesis in vitro: role of extracellular matrix. J Cell Biol. 1989 Jul;109(1):317–330. doi: 10.1083/jcb.109.1.317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ingber D. E., Madri J. A., Folkman J. Endothelial growth factors and extracellular matrix regulate DNA synthesis through modulation of cell and nuclear expansion. In Vitro Cell Dev Biol. 1987 May;23(5):387–394. doi: 10.1007/BF02620997. [DOI] [PubMed] [Google Scholar]
  28. Ingber D. Extracellular matrix and cell shape: potential control points for inhibition of angiogenesis. J Cell Biochem. 1991 Nov;47(3):236–241. doi: 10.1002/jcb.240470309. [DOI] [PubMed] [Google Scholar]
  29. Ingber D., Folkman J. Inhibition of angiogenesis through modulation of collagen metabolism. Lab Invest. 1988 Jul;59(1):44–51. [PubMed] [Google Scholar]
  30. Ingber D. Integrins as mechanochemical transducers. Curr Opin Cell Biol. 1991 Oct;3(5):841–848. doi: 10.1016/0955-0674(91)90058-7. [DOI] [PubMed] [Google Scholar]
  31. Iruela-Arispe M. L., Hasselaar P., Sage H. Differential expression of extracellular proteins is correlated with angiogenesis in vitro. Lab Invest. 1991 Feb;64(2):174–186. [PubMed] [Google Scholar]
  32. Kornberg L. J., Earp H. S., Turner C. E., Prockop C., Juliano R. L. Signal transduction by integrins: increased protein tyrosine phosphorylation caused by clustering of beta 1 integrins. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8392–8396. doi: 10.1073/pnas.88.19.8392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kramer R. H., Cheng Y. F., Clyman R. Human microvascular endothelial cells use beta 1 and beta 3 integrin receptor complexes to attach to laminin. J Cell Biol. 1990 Sep;111(3):1233–1243. doi: 10.1083/jcb.111.3.1233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Kubota Y., Kleinman H. K., Martin G. R., Lawley T. J. Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-like structures. J Cell Biol. 1988 Oct;107(4):1589–1598. doi: 10.1083/jcb.107.4.1589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Languino L. R., Gehlsen K. R., Wayner E., Carter W. G., Engvall E., Ruoslahti E. Endothelial cells use alpha 2 beta 1 integrin as a laminin receptor. J Cell Biol. 1989 Nov;109(5):2455–2462. doi: 10.1083/jcb.109.5.2455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Madri J. A., Pratt B. M., Tucker A. M. Phenotypic modulation of endothelial cells by transforming growth factor-beta depends upon the composition and organization of the extracellular matrix. J Cell Biol. 1988 Apr;106(4):1375–1384. doi: 10.1083/jcb.106.4.1375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Mallein-Gerin F., Garrone R., van der Rest M. Proteoglycan and collagen synthesis are correlated with actin organization in dedifferentiating chondrocytes. Eur J Cell Biol. 1991 Dec;56(2):364–373. [PubMed] [Google Scholar]
  38. Marks R. M., Czerniecki M., Penny R. Human dermal microvascular endothelial cells: an improved method for tissue culture and a description of some singular properties in culture. In Vitro Cell Dev Biol. 1985 Nov;21(11):627–635. doi: 10.1007/BF02623295. [DOI] [PubMed] [Google Scholar]
  39. Mazurov A. V., Vinogradov D. V., Kabaeva N. V., Antonova G. N., Romanov Y. A., Vlasik T. N., Antonov A. S., Smirnov V. N. A monoclonal antibody, VM64, reacts with a 130 kDa glycoprotein common to platelets and endothelial cells: heterogeneity in antibody binding to human aortic endothelial cells. Thromb Haemost. 1991 Oct 1;66(4):494–499. [PubMed] [Google Scholar]
  40. Mignatti P., Mazzieri R., Rifkin D. B. Expression of the urokinase receptor in vascular endothelial cells is stimulated by basic fibroblast growth factor. J Cell Biol. 1991 Jun;113(5):1193–1201. doi: 10.1083/jcb.113.5.1193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Montesano R., Mouron P., Orci L. Vascular outgrowths from tissue explants embedded in fibrin or collagen gels: a simple in vitro model of angiogenesis. Cell Biol Int Rep. 1985 Oct;9(10):869–875. doi: 10.1016/s0309-1651(85)90107-9. [DOI] [PubMed] [Google Scholar]
  42. Montesano R., Orci L. Tumor-promoting phorbol esters induce angiogenesis in vitro. Cell. 1985 Sep;42(2):469–477. doi: 10.1016/0092-8674(85)90104-7. [DOI] [PubMed] [Google Scholar]
  43. Montesano R., Orci L., Vassalli P. In vitro rapid organization of endothelial cells into capillary-like networks is promoted by collagen matrices. J Cell Biol. 1983 Nov;97(5 Pt 1):1648–1652. doi: 10.1083/jcb.97.5.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Montesano R., Pepper M. S., Vassalli J. D., Orci L. Phorbol ester induces cultured endothelial cells to invade a fibrin matrix in the presence of fibrinolytic inhibitors. J Cell Physiol. 1987 Sep;132(3):509–516. doi: 10.1002/jcp.1041320313. [DOI] [PubMed] [Google Scholar]
  45. Nethery A., Lyons J. G., O'Grady R. L. A spectrophotometric collagenase assay. Anal Biochem. 1986 Dec;159(2):390–395. doi: 10.1016/0003-2697(86)90358-1. [DOI] [PubMed] [Google Scholar]
  46. O'Connell P. J., Faull R., Russ G. R., D'Apice A. J. VLA-2 is a collagen receptor on endothelial cells. Immunol Cell Biol. 1991 Apr;69(Pt 2):103–110. doi: 10.1038/icb.1991.16. [DOI] [PubMed] [Google Scholar]
  47. Oliver M. H., Harrison N. K., Bishop J. E., Cole P. J., Laurent G. J. A rapid and convenient assay for counting cells cultured in microwell plates: application for assessment of growth factors. J Cell Sci. 1989 Mar;92(Pt 3):513–518. doi: 10.1242/jcs.92.3.513. [DOI] [PubMed] [Google Scholar]
  48. Ruoslahti E., Pierschbacher M. D. New perspectives in cell adhesion: RGD and integrins. Science. 1987 Oct 23;238(4826):491–497. doi: 10.1126/science.2821619. [DOI] [PubMed] [Google Scholar]
  49. Sholley M. M., Ferguson G. P., Seibel H. R., Montour J. L., Wilson J. D. Mechanisms of neovascularization. Vascular sprouting can occur without proliferation of endothelial cells. Lab Invest. 1984 Dec;51(6):624–634. [PubMed] [Google Scholar]
  50. Solowska J., Guan J. L., Marcantonio E. E., Trevithick J. E., Buck C. A., Hynes R. O. Expression of normal and mutant avian integrin subunits in rodent cells. J Cell Biol. 1989 Aug;109(2):853–861. doi: 10.1083/jcb.109.2.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Unemori E. N., Werb Z. Reorganization of polymerized actin: a possible trigger for induction of procollagenase in fibroblasts cultured in and on collagen gels. J Cell Biol. 1986 Sep;103(3):1021–1031. doi: 10.1083/jcb.103.3.1021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Wall R. T., Harker L. A., Quadracci L. J., Striker G. E. Factors influencing endothelial cell proliferation in vitro. J Cell Physiol. 1978 Aug;96(2):203–213. doi: 10.1002/jcp.1040960209. [DOI] [PubMed] [Google Scholar]
  53. Watson P. A. Function follows form: generation of intracellular signals by cell deformation. FASEB J. 1991 Apr;5(7):2013–2019. doi: 10.1096/fasebj.5.7.1707019. [DOI] [PubMed] [Google Scholar]
  54. Werb Z., Hembry R. M., Murphy G., Aggeler J. Commitment to expression of the metalloendopeptidases, collagenase and stromelysin: relationship of inducing events to changes in cytoskeletal architecture. J Cell Biol. 1986 Mar;102(3):697–702. doi: 10.1083/jcb.102.3.697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Werb Z., Tremble P. M., Behrendtsen O., Crowley E., Damsky C. H. Signal transduction through the fibronectin receptor induces collagenase and stromelysin gene expression. J Cell Biol. 1989 Aug;109(2):877–889. doi: 10.1083/jcb.109.2.877. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES