Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 Sep 1;98(5):1133–1141. doi: 10.1172/JCI118896

Integrin-dependent induction of functional urokinase receptors in primary T lymphocytes.

E Bianchi 1, E Ferrero 1, F Fazioli 1, F Mangili 1, J Wang 1, J R Bender 1, F Blasi 1, R Pardi 1
PMCID: PMC507535  PMID: 8787676

Abstract

In order to reach the sites of inflammation, lymphocytes leave the bloodstream and migrate into peripheral tissues, in a process involving integrin-mediated adhesion to the vascular endothelium, followed by transmigration across the endothelial barrier and through the underlying interstitial matrix. We have investigated the role of the plasminogen activator/plasmin system in normal T cell migration. Receptors for urokinase plasminogen activator (uPAR) were not expressed in resting T lymphocytes, but could be efficiently induced at the mRNA and protein level by coclustering of the antigen receptor complex and beta1 or beta2 integrins, through a signalling pathway involving both protein kinase C activation and an increase in intracellular cyclic AMP. Catalytic activation of plasminogen by uPAR-expressing T cells promoted their migration through an extracellular matrix in vitro. Plasmin-induced invasion was inhibited by plasmin-and urokinase inhibitors and by anti-uPAR antibodies. Finally, cytofluorimetric and immunohistochemical analysis of primary human tumor specimens showed the presence of uPAR positive infiltrating T cells in vivo. Collectively, these findings suggest that plasminogen activation may play a role in lymphocyte migration in vivo, and that integrin-dependent expression of membrane-associated endopeptidases could represent an additional step in the regulated process of leukocyte transmigration.

Full Text

The Full Text of this article is available as a PDF (481.1 KB).

Selected References

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

  1. Appella E., Robinson E. A., Ullrich S. J., Stoppelli M. P., Corti A., Cassani G., Blasi F. The receptor-binding sequence of urokinase. A biological function for the growth-factor module of proteases. J Biol Chem. 1987 Apr 5;262(10):4437–4440. [PubMed] [Google Scholar]
  2. Bianchi E., Cohen R. L., Thor A. T., Todd R. F., 3rd, Mizukami I. F., Lawrence D. A., Ljung B. M., Shuman M. A., Smith H. S. The urokinase receptor is expressed in invasive breast cancer but not in normal breast tissue. Cancer Res. 1994 Feb 15;54(4):861–866. [PubMed] [Google Scholar]
  3. Bristow C. L., Lyford L. K., Stevens D. P., Flood P. M. Elastase is a constituent product of T cells. Biochem Biophys Res Commun. 1991 Nov 27;181(1):232–239. doi: 10.1016/s0006-291x(05)81407-x. [DOI] [PubMed] [Google Scholar]
  4. Clipstone N. A., Crabtree G. R. Calcineurin is a key signaling enzyme in T lymphocyte activation and the target of the immunosuppressive drugs cyclosporin A and FK506. Ann N Y Acad Sci. 1993 Nov 30;696:20–30. doi: 10.1111/j.1749-6632.1993.tb17138.x. [DOI] [PubMed] [Google Scholar]
  5. Conese M., Blasi F. The urokinase/urokinase-receptor system and cancer invasion. Baillieres Clin Haematol. 1995 Jun;8(2):365–389. doi: 10.1016/s0950-3536(05)80273-2. [DOI] [PubMed] [Google Scholar]
  6. Crowley C. W., Cohen R. L., Lucas B. K., Liu G., Shuman M. A., Levinson A. D. Prevention of metastasis by inhibition of the urokinase receptor. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5021–5025. doi: 10.1073/pnas.90.11.5021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dustin M. L., Springer T. A. T-cell receptor cross-linking transiently stimulates adhesiveness through LFA-1. Nature. 1989 Oct 19;341(6243):619–624. doi: 10.1038/341619a0. [DOI] [PubMed] [Google Scholar]
  8. Estreicher A., Mühlhauser J., Carpentier J. L., Orci L., Vassalli J. D. The receptor for urokinase type plasminogen activator polarizes expression of the protease to the leading edge of migrating monocytes and promotes degradation of enzyme inhibitor complexes. J Cell Biol. 1990 Aug;111(2):783–792. doi: 10.1083/jcb.111.2.783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fulton R. J., Hart D. A. Detection and partial characterization of lymphoid cell surface proteases. Cell Immunol. 1980 Oct;55(2):394–405. doi: 10.1016/0008-8749(80)90170-7. [DOI] [PubMed] [Google Scholar]
  10. Genot E. M., Parker P. J., Cantrell D. A. Analysis of the role of protein kinase C-alpha, -epsilon, and -zeta in T cell activation. J Biol Chem. 1995 Apr 28;270(17):9833–9839. doi: 10.1074/jbc.270.17.9833. [DOI] [PubMed] [Google Scholar]
  11. Hearing V. J., Law L. W., Corti A., Appella E., Blasi F. Modulation of metastatic potential by cell surface urokinase of murine melanoma cells. Cancer Res. 1988 Mar 1;48(5):1270–1278. [PubMed] [Google Scholar]
  12. Imboden J. B., Stobo J. D. Transmembrane signalling by the T cell antigen receptor. Perturbation of the T3-antigen receptor complex generates inositol phosphates and releases calcium ions from intracellular stores. J Exp Med. 1985 Mar 1;161(3):446–456. doi: 10.1084/jem.161.3.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Imhof B. A., Dunon D. Leukocyte migration and adhesion. Adv Immunol. 1995;58:345–416. doi: 10.1016/s0065-2776(08)60623-9. [DOI] [PubMed] [Google Scholar]
  14. Kanner S. B., Grosmaire L. S., Ledbetter J. A., Damle N. K. Beta 2-integrin LFA-1 signaling through phospholipase C-gamma 1 activation. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7099–7103. doi: 10.1073/pnas.90.15.7099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kobayashi H., Gotoh J., Fujie M., Shinohara H., Moniwa N., Terao T. Inhibition of metastasis of Lewis lung carcinoma by a synthetic peptide within growth factor-like domain of urokinase in the experimental and spontaneous metastasis model. Int J Cancer. 1994 Jun 1;57(5):727–733. doi: 10.1002/ijc.2910570520. [DOI] [PubMed] [Google Scholar]
  16. Kramer M. D., Schaefer B., Reinartz J. Plasminogen activation by human keratinocytes: molecular pathways and cell-biological consequences. Biol Chem Hoppe Seyler. 1995 Mar;376(3):131–141. doi: 10.1515/bchm3.1995.376.3.131. [DOI] [PubMed] [Google Scholar]
  17. Kramer M. D., Spring H., Todd R. F., Vettel U. Urokinase-type plasminogen activator enhances invasion of human T cells (Jurkat) into a fibrin matrix. J Leukoc Biol. 1994 Aug;56(2):110–116. doi: 10.1002/jlb.56.2.110. [DOI] [PubMed] [Google Scholar]
  18. Langer D. J., Kuo A., Kariko K., Ahuja M., Klugherz B. D., Ivanics K. M., Hoxie J. A., Williams W. V., Liang B. T., Cines D. B. Regulation of the endothelial cell urokinase-type plasminogen activator receptor. Evidence for cyclic AMP-dependent and protein kinase C-dependent pathways. Circ Res. 1993 Feb;72(2):330–340. doi: 10.1161/01.res.72.2.330. [DOI] [PubMed] [Google Scholar]
  19. Larjava H., Lyons J. G., Salo T., Mäkelä M., Koivisto L., Birkedal-Hansen H., Akiyama S. K., Yamada K. M., Heino J. Anti-integrin antibodies induce type IV collagenase expression in keratinocytes. J Cell Physiol. 1993 Oct;157(1):190–200. doi: 10.1002/jcp.1041570125. [DOI] [PubMed] [Google Scholar]
  20. Leppert D., Waubant E., Galardy R., Bunnett N. W., Hauser S. L. T cell gelatinases mediate basement membrane transmigration in vitro. J Immunol. 1995 May 1;154(9):4379–4389. [PubMed] [Google Scholar]
  21. Mignatti P., Robbins E., Rifkin D. B. Tumor invasion through the human amniotic membrane: requirement for a proteinase cascade. Cell. 1986 Nov 21;47(4):487–498. doi: 10.1016/0092-8674(86)90613-6. [DOI] [PubMed] [Google Scholar]
  22. Miles L. A., Plow E. F. Receptor mediated binding of the fibrinolytic components, plasminogen and urokinase, to peripheral blood cells. Thromb Haemost. 1987 Oct 28;58(3):936–942. [PubMed] [Google Scholar]
  23. Min H. Y., Semnani R., Mizukami I. F., Watt K., Todd R. F., 3rd, Liu D. Y. cDNA for Mo3, a monocyte activation antigen, encodes the human receptor for urokinase plasminogen activator. J Immunol. 1992 Jun 1;148(11):3636–3642. [PubMed] [Google Scholar]
  24. Nykjaer A., Møller B., Todd R. F., 3rd, Christensen T., Andreasen P. A., Gliemann J., Petersen C. M. Urokinase receptor. An activation antigen in human T lymphocytes. J Immunol. 1994 Jan 15;152(2):505–516. [PubMed] [Google Scholar]
  25. Nykjaer A., Petersen C. M., Møller B., Andreasen P. A., Gliemann J. Identification and characterization of urokinase receptors in natural killer cells and T-cell-derived lymphokine activated killer cells. FEBS Lett. 1992 Mar 23;300(1):13–17. doi: 10.1016/0014-5793(92)80154-9. [DOI] [PubMed] [Google Scholar]
  26. Ossowski L. In vivo invasion of modified chorioallantoic membrane by tumor cells: the role of cell surface-bound urokinase. J Cell Biol. 1988 Dec;107(6 Pt 1):2437–2445. doi: 10.1083/jcb.107.6.2437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ossowski L., Reich E. Antibodies to plasminogen activator inhibit human tumor metastasis. Cell. 1983 Dec;35(3 Pt 2):611–619. doi: 10.1016/0092-8674(83)90093-4. [DOI] [PubMed] [Google Scholar]
  28. Ossowski L., Russo-Payne H., Wilson E. L. Inhibition of urokinase-type plasminogen activator by antibodies: the effect on dissemination of a human tumor in the nude mouse. Cancer Res. 1991 Jan 1;51(1):274–281. [PubMed] [Google Scholar]
  29. Pardi R., Bender J. R., Dettori C., Giannazza E., Engleman E. G. Heterogeneous distribution and transmembrane signaling properties of lymphocyte function-associated antigen (LFA-1) in human lymphocyte subsets. J Immunol. 1989 Nov 15;143(10):3157–3166. [PubMed] [Google Scholar]
  30. Pardi R., Inverardi L., Rugarli C., Bender J. R. Antigen-receptor complex stimulation triggers protein kinase C-dependent CD11a/CD18-cytoskeleton association in T lymphocytes. J Cell Biol. 1992 Mar;116(5):1211–1220. doi: 10.1083/jcb.116.5.1211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Picker L. J., Butcher E. C. Physiological and molecular mechanisms of lymphocyte homing. Annu Rev Immunol. 1992;10:561–591. doi: 10.1146/annurev.iy.10.040192.003021. [DOI] [PubMed] [Google Scholar]
  32. Pyke C., Graem N., Ralfkiaer E., Rønne E., Høyer-Hansen G., Brünner N., Danø K. Receptor for urokinase is present in tumor-associated macrophages in ductal breast carcinoma. Cancer Res. 1993 Apr 15;53(8):1911–1915. [PubMed] [Google Scholar]
  33. Pyke C., Kristensen P., Ralfkiaer E., Grøndahl-Hansen J., Eriksen J., Blasi F., Danø K. Urokinase-type plasminogen activator is expressed in stromal cells and its receptor in cancer cells at invasive foci in human colon adenocarcinomas. Am J Pathol. 1991 May;138(5):1059–1067. [PMC free article] [PubMed] [Google Scholar]
  34. Resnati M., Guttinger M., Valcamonica S., Sidenius N., Blasi F., Fazioli F. Proteolytic cleavage of the urokinase receptor substitutes for the agonist-induced chemotactic effect. EMBO J. 1996 Apr 1;15(7):1572–1582. [PMC free article] [PubMed] [Google Scholar]
  35. Roldan A. L., Cubellis M. V., Masucci M. T., Behrendt N., Lund L. R., Danø K., Appella E., Blasi F. Cloning and expression of the receptor for human urokinase plasminogen activator, a central molecule in cell surface, plasmin dependent proteolysis. EMBO J. 1990 Feb;9(2):467–474. doi: 10.1002/j.1460-2075.1990.tb08132.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Romanic A. M., Madri J. A. The induction of 72-kD gelatinase in T cells upon adhesion to endothelial cells is VCAM-1 dependent. J Cell Biol. 1994 Jun;125(5):1165–1178. doi: 10.1083/jcb.125.5.1165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rosales C., Juliano R. L. Signal transduction by cell adhesion receptors in leukocytes. J Leukoc Biol. 1995 Feb;57(2):189–198. doi: 10.1002/jlb.57.2.189. [DOI] [PubMed] [Google Scholar]
  38. Rovere P., Inverardi L., Bender J. R., Pardi R. Feedback modulation of ligand-engaged alpha L/beta 2 leukocyte integrin (LFA-1) by cyclic AMP-dependent protein kinase. J Immunol. 1996 Mar 15;156(6):2273–2279. [PubMed] [Google Scholar]
  39. Rønne E., Behrendt N., Ellis V., Ploug M., Danø K., Høyer-Hansen G. Cell-induced potentiation of the plasminogen activation system is abolished by a monoclonal antibody that recognizes the NH2-terminal domain of the urokinase receptor. FEBS Lett. 1991 Aug 19;288(1-2):233–236. doi: 10.1016/0014-5793(91)81042-7. [DOI] [PubMed] [Google Scholar]
  40. Seftor R. E., Seftor E. A., Stetler-Stevenson W. G., Hendrix M. J. The 72 kDa type IV collagenase is modulated via differential expression of alpha v beta 3 and alpha 5 beta 1 integrins during human melanoma cell invasion. Cancer Res. 1993 Jul 15;53(14):3411–3415. [PubMed] [Google Scholar]
  41. Siebenlist U., Franzoso G., Brown K. Structure, regulation and function of NF-kappa B. Annu Rev Cell Biol. 1994;10:405–455. doi: 10.1146/annurev.cb.10.110194.002201. [DOI] [PubMed] [Google Scholar]
  42. Soravia E., Grebe A., De Luca P., Helin K., Suh T. T., Degen J. L., Blasi F. A conserved TATA-less proximal promoter drives basal transcription from the urokinase-type plasminogen activator receptor gene. Blood. 1995 Jul 15;86(2):624–635. [PubMed] [Google Scholar]
  43. Springer T. A. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994 Jan 28;76(2):301–314. doi: 10.1016/0092-8674(94)90337-9. [DOI] [PubMed] [Google Scholar]
  44. Stoppelli M. P., Corti A., Soffientini A., Cassani G., Blasi F., Assoian R. K. Differentiation-enhanced binding of the amino-terminal fragment of human urokinase plasminogen activator to a specific receptor on U937 monocytes. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4939–4943. doi: 10.1073/pnas.82.15.4939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Szamel M., Resch K. T-cell antigen receptor-induced signal-transduction pathways--activation and function of protein kinases C in T lymphocytes. Eur J Biochem. 1995 Feb 15;228(1):1–15. doi: 10.1111/j.1432-1033.1995.tb20221.x. [DOI] [PubMed] [Google Scholar]
  46. Vassalli J. D., Baccino D., Belin D. A cellular binding site for the Mr 55,000 form of the human plasminogen activator, urokinase. J Cell Biol. 1985 Jan;100(1):86–92. doi: 10.1083/jcb.100.1.86. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Vassalli J. D., Belin D. Amiloride selectively inhibits the urokinase-type plasminogen activator. FEBS Lett. 1987 Apr 6;214(1):187–191. doi: 10.1016/0014-5793(87)80039-x. [DOI] [PubMed] [Google Scholar]
  48. Vassalli J. D., Sappino A. P., Belin D. The plasminogen activator/plasmin system. J Clin Invest. 1991 Oct;88(4):1067–1072. doi: 10.1172/JCI115405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Vlodavsky I., Eldor A., Haimovitz-Friedman A., Matzner Y., Ishai-Michaeli R., Lider O., Naparstek Y., Cohen I. R., Fuks Z. Expression of heparanase by platelets and circulating cells of the immune system: possible involvement in diapedesis and extravasation. Invasion Metastasis. 1992;12(2):112–127. [PubMed] [Google Scholar]
  50. Waltz D. A., Chapman H. A. Reversible cellular adhesion to vitronectin linked to urokinase receptor occupancy. J Biol Chem. 1994 May 20;269(20):14746–14750. [PubMed] [Google Scholar]
  51. 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]
  52. Wysocki L. J., Sato V. L. "Panning" for lymphocytes: a method for cell selection. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2844–2848. doi: 10.1073/pnas.75.6.2844. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES