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. 1995 May 1;181(5):1805–1815. doi: 10.1084/jem.181.5.1805

Cytokines increase human hemopoietic cell adhesiveness by activation of very late antigen (VLA)-4 and VLA-5 integrins

PMCID: PMC2192007  PMID: 7536795

Abstract

Cytokines are known to be important regulators of normal hemopoiesis, acting in concert with components of the bone marrow microenvironment. Interactions with this microenvironment are known to regulate the proliferation, differentiation, and homing of hemopoietic progenitor (CD34+) cells. Adhesive interactions with the extracellular matrix retain CD34+ cells in close proximity to cytokines, but may also provide important costimulatory signals. Thus, the functional states of adhesion receptors are critical properties of CD34+ cells, but the physiological mechanisms responsible for regulating functional properties of cell adhesion receptors on primitive hemopoietic cells are still unknown. We confirm that the integrins very late antigen (VLA)-4 and VLA-5 are expressed on the CD34+ cell lines MO7e, TF1, and on normal bone marrow CD34+ progenitor cells, but in a low affinity state, conferring on them a weak adhesive phenotype on fibronectin (Fn). Herein, we show that the cytokines interleukin (IL)-3, granulocyte-macrophage CSF (GM-CSF), and KIT ligand (KL) are physiological activators of VLA-4 and VLA-5 expressed by MO7e, TF1, and normal bone marrow CD34+ progenitor cells. Cytokine-stimulated adhesion on Fn is dose dependent and transient, reaching a maximum between 15 and 30 min and returning to basal levels after 2 h. This cytokine- dependent activation is specific for VLA-4 and VLA-5, since activation of other beta 1 integrins was not observed. The addition of second messenger antagonists staurosporine and W7 abolished all cytokine- stimulated adhesion to Fn. In contrast, genistein inhibited KL- stimulated adhesion, but failed to inhibit GM-CSF- and IL-3-stimulated adhesion. Our data suggest that cytokines GM-CSF and IL-3 specifically stimulate beta 1 integrin function via an "inside-out" mechanism involving protein kinase activity, while KL stimulates integrin activity through a similar, but initially distinct, pathway via the KIT tyrosine-kinase. Thus, in addition to promoting the survival, proliferation, and development of hemopoietic progenitors, cytokines also regulate adhesive interactions between progenitor cells and the bone marrow microenvironment by modifying the functional states of specific integrins. These data are of importance in understanding the fundamental processes of beta 1 integrin activation and cellular response to mitogenic cytokines as well as on the clinical setting where cytokines induce therapeutic mobilization of hematopoietic progenitors.

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Selected References

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  1. Andrews R. G., Singer J. W., Bernstein I. D. Precursors of colony-forming cells in humans can be distinguished from colony-forming cells by expression of the CD33 and CD34 antigens and light scatter properties. J Exp Med. 1989 May 1;169(5):1721–1731. doi: 10.1084/jem.169.5.1721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arroyo A. G., García-Pardo A., Sánchez-Madrid F. A high affinity conformational state on VLA integrin heterodimers induced by an anti-beta 1 chain monoclonal antibody. J Biol Chem. 1993 May 5;268(13):9863–9868. [PubMed] [Google Scholar]
  3. Ashman L. K., Cambareri A. C., To L. B., Levinsky R. J., Juttner C. A. Expression of the YB5.B8 antigen (c-kit proto-oncogene product) in normal human bone marrow. Blood. 1991 Jul 1;78(1):30–37. [PubMed] [Google Scholar]
  4. Avanzi G. C., Lista P., Giovinazzo B., Miniero R., Saglio G., Benetton G., Coda R., Cattoretti G., Pegoraro L. Selective growth response to IL-3 of a human leukaemic cell line with megakaryoblastic features. Br J Haematol. 1988 Jul;69(3):359–366. doi: 10.1111/j.1365-2141.1988.tb02374.x. [DOI] [PubMed] [Google Scholar]
  5. Barovsky K., Brooker G. Forskolin potentiation of cholera toxin-stimulated cyclic AMP accumulation in intact C6-2B cells. Evidence for enhanced Gs-C coupling. Mol Pharmacol. 1985 Dec;28(6):502–507. [PubMed] [Google Scholar]
  6. Bohnsack J. F., Chang J. Activation of beta 1 integrin fibronectin receptors on HL60 cells after granulocytic differentiation. Blood. 1994 Jan 15;83(2):543–552. [PubMed] [Google Scholar]
  7. Corey S., Eguinoa A., Puyana-Theall K., Bolen J. B., Cantley L., Mollinedo F., Jackson T. R., Hawkins P. T., Stephens L. R. Granulocyte macrophage-colony stimulating factor stimulates both association and activation of phosphoinositide 3OH-kinase and src-related tyrosine kinase(s) in human myeloid derived cells. EMBO J. 1993 Jul;12(7):2681–2690. doi: 10.1002/j.1460-2075.1993.tb05929.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Danilov Y. N., Juliano R. L. Phorbol ester modulation of integrin-mediated cell adhesion: a postreceptor event. J Cell Biol. 1989 May;108(5):1925–1933. doi: 10.1083/jcb.108.5.1925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dexter T. M., Allen T. D., Lajtha L. G. Conditions controlling the proliferation of haemopoietic stem cells in vitro. J Cell Physiol. 1977 Jun;91(3):335–344. doi: 10.1002/jcp.1040910303. [DOI] [PubMed] [Google Scholar]
  10. Faull R. J., Kovach N. L., Harlan J. M., Ginsberg M. H. Affinity modulation of integrin alpha 5 beta 1: regulation of the functional response by soluble fibronectin. J Cell Biol. 1993 Apr;121(1):155–162. doi: 10.1083/jcb.121.1.155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Faull R. J., Kovach N. L., Harlan J. M., Ginsberg M. H. Stimulation of integrin-mediated adhesion of T lymphocytes and monocytes: two mechanisms with divergent biological consequences. J Exp Med. 1994 Apr 1;179(4):1307–1316. doi: 10.1084/jem.179.4.1307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ginsberg M. H., Du X., Plow E. F. Inside-out integrin signalling. Curr Opin Cell Biol. 1992 Oct;4(5):766–771. doi: 10.1016/0955-0674(92)90099-x. [DOI] [PubMed] [Google Scholar]
  13. Ginsberg M. H., O'Toole T. E., Loftus J. C., Plow E. F. Ligand binding to integrins: dynamic regulation and common mechanisms. Cold Spring Harb Symp Quant Biol. 1992;57:221–231. doi: 10.1101/sqb.1992.057.01.027. [DOI] [PubMed] [Google Scholar]
  14. Haystead T. A., Sim A. T., Carling D., Honnor R. C., Tsukitani Y., Cohen P., Hardie D. G. Effects of the tumour promoter okadaic acid on intracellular protein phosphorylation and metabolism. Nature. 1989 Jan 5;337(6202):78–81. doi: 10.1038/337078a0. [DOI] [PubMed] [Google Scholar]
  15. Hibbs M. L., Xu H., Stacker S. A., Springer T. A. Regulation of adhesion of ICAM-1 by the cytoplasmic domain of LFA-1 integrin beta subunit. Science. 1991 Mar 29;251(5001):1611–1613. doi: 10.1126/science.1672776. [DOI] [PubMed] [Google Scholar]
  16. Hynes R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. doi: 10.1016/0092-8674(92)90115-s. [DOI] [PubMed] [Google Scholar]
  17. Itoh H., Hidaka H. Direct interaction of calmodulin antagonists with Ca2+/calmodulin-dependent cyclic nucleotide phosphodiesterase. J Biochem. 1984 Dec;96(6):1721–1726. doi: 10.1093/oxfordjournals.jbchem.a135004. [DOI] [PubMed] [Google Scholar]
  18. Kerst J. M., Sanders J. B., Slaper-Cortenbach I. C., Doorakkers M. C., Hooibrink B., van Oers R. H., von dem Borne A. E., van der Schoot C. E. Alpha 4 beta 1 and alpha 5 beta 1 are differentially expressed during myelopoiesis and mediate the adherence of human CD34+ cells to fibronectin in an activation-dependent way. Blood. 1993 Jan 15;81(2):344–351. [PubMed] [Google Scholar]
  19. Kitamura T., Tange T., Terasawa T., Chiba S., Kuwaki T., Miyagawa K., Piao Y. F., Miyazono K., Urabe A., Takaku F. Establishment and characterization of a unique human cell line that proliferates dependently on GM-CSF, IL-3, or erythropoietin. J Cell Physiol. 1989 Aug;140(2):323–334. doi: 10.1002/jcp.1041400219. [DOI] [PubMed] [Google Scholar]
  20. Kovach N. L., Carlos T. M., Yee E., Harlan J. M. A monoclonal antibody to beta 1 integrin (CD29) stimulates VLA-dependent adherence of leukocytes to human umbilical vein endothelial cells and matrix components. J Cell Biol. 1992 Jan;116(2):499–509. doi: 10.1083/jcb.116.2.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Leavesley D. I., Oliver J. M., Swart B. W., Berndt M. C., Haylock D. N., Simmons P. J. Signals from platelet/endothelial cell adhesion molecule enhance the adhesive activity of the very late antigen-4 integrin of human CD34+ hemopoietic progenitor cells. J Immunol. 1994 Nov 15;153(10):4673–4683. [PubMed] [Google Scholar]
  22. Makrynikola V., Bradstock K. F. Adhesion of precursor-B acute lymphoblastic leukaemia cells to bone marrow stromal proteins. Leukemia. 1993 Jan;7(1):86–92. [PubMed] [Google Scholar]
  23. Masumoto A., Hemler M. E. Multiple activation states of VLA-4. Mechanistic differences between adhesion to CS1/fibronectin and to vascular cell adhesion molecule-1. J Biol Chem. 1993 Jan 5;268(1):228–234. [PubMed] [Google Scholar]
  24. Metcalf D. Hematopoietic regulators: redundancy or subtlety? Blood. 1993 Dec 15;82(12):3515–3523. [PubMed] [Google Scholar]
  25. Miyake K., Weissman I. L., Greenberger J. S., Kincade P. W. Evidence for a role of the integrin VLA-4 in lympho-hemopoiesis. J Exp Med. 1991 Mar 1;173(3):599–607. doi: 10.1084/jem.173.3.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. O'Toole T. E., Mandelman D., Forsyth J., Shattil S. J., Plow E. F., Ginsberg M. H. Modulation of the affinity of integrin alpha IIb beta 3 (GPIIb-IIIa) by the cytoplasmic domain of alpha IIb. Science. 1991 Nov 8;254(5033):845–847. doi: 10.1126/science.1948065. [DOI] [PubMed] [Google Scholar]
  27. Okuda K., Sanghera J. S., Pelech S. L., Kanakura Y., Hallek M., Griffin J. D., Druker B. J. Granulocyte-macrophage colony-stimulating factor, interleukin-3, and steel factor induce rapid tyrosine phosphorylation of p42 and p44 MAP kinase. Blood. 1992 Jun 1;79(11):2880–2887. [PubMed] [Google Scholar]
  28. Platanias L. C., Colamonici O. R. Interferon alpha induces rapid tyrosine phosphorylation of the alpha subunit of its receptor. J Biol Chem. 1992 Nov 25;267(33):24053–24057. [PubMed] [Google Scholar]
  29. Quelle F. W., Sato N., Witthuhn B. A., Inhorn R. C., Eder M., Miyajima A., Griffin J. D., Ihle J. N. JAK2 associates with the beta c chain of the receptor for granulocyte-macrophage colony-stimulating factor, and its activation requires the membrane-proximal region. Mol Cell Biol. 1994 Jul;14(7):4335–4341. doi: 10.1128/mcb.14.7.4335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rosemblatt M., Vuillet-Gaugler M. H., Leroy C., Coulombel L. Coexpression of two fibronectin receptors, VLA-4 and VLA-5, by immature human erythroblastic precursor cells. J Clin Invest. 1991 Jan;87(1):6–11. doi: 10.1172/JCI115002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ryan D. H., Nuccie B. L., Abboud C. N., Winslow J. M. Vascular cell adhesion molecule-1 and the integrin VLA-4 mediate adhesion of human B cell precursors to cultured bone marrow adherent cells. J Clin Invest. 1991 Sep;88(3):995–1004. doi: 10.1172/JCI115403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sato N., Sawada K., Takahashi T. A., Mogi Y., Asano S., Koike T., Sekiguchi S. A time course study for optimal harvest of peripheral blood progenitor cells by granulocyte colony-stimulating factor in healthy volunteers. Exp Hematol. 1994 Sep;22(10):973–978. [PubMed] [Google Scholar]
  33. Shaw L. M., Mercurio A. M. Regulation of alpha 6 beta 1 integrin laminin receptor function by the cytoplasmic domain of the alpha 6 subunit. J Cell Biol. 1993 Nov;123(4):1017–1025. doi: 10.1083/jcb.123.4.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shaw L. M., Messier J. M., Mercurio A. M. The activation dependent adhesion of macrophages to laminin involves cytoskeletal anchoring and phosphorylation of the alpha 6 beta 1 integrin. J Cell Biol. 1990 Jun;110(6):2167–2174. doi: 10.1083/jcb.110.6.2167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Shimizu Y., van Seventer G. A., Ennis E., Newman W., Horgan K. J., Shaw S. Crosslinking of the T cell-specific accessory molecules CD7 and CD28 modulates T cell adhesion. J Exp Med. 1992 Feb 1;175(2):577–582. doi: 10.1084/jem.175.2.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Simmons P. J., Leavesley D. I., Levesque J. P., Swart B. W., Haylock D. N., To L. B., Ashman L. K., Juttner C. A. The mobilization of primitive hemopoietic progenitors into the peripheral blood. Stem Cells. 1994;12 (Suppl 1):187–202. [PubMed] [Google Scholar]
  37. Simmons P. J., Zannettino A., Gronthos S., Leavesley D. Potential adhesion mechanisms for localisation of haemopoietic progenitors to bone marrow stroma. Leuk Lymphoma. 1994 Feb;12(5-6):353–363. doi: 10.3109/10428199409073776. [DOI] [PubMed] [Google Scholar]
  38. Soligo D., Schiró R., Luksch R., Manara G., Quirici N., Parravicini C., Lambertenghi Deliliers G. Expression of integrins in human bone marrow. Br J Haematol. 1990 Nov;76(3):323–332. doi: 10.1111/j.1365-2141.1990.tb06363.x. [DOI] [PubMed] [Google Scholar]
  39. Suganuma M., Fujiki H., Furuya-Suguri H., Yoshizawa S., Yasumoto S., Kato Y., Fusetani N., Sugimura T. Calyculin A, an inhibitor of protein phosphatases, a potent tumor promoter on CD-1 mouse skin. Cancer Res. 1990 Jun 15;50(12):3521–3525. [PubMed] [Google Scholar]
  40. Takada Y., Puzon W. Identification of a regulatory region of integrin beta 1 subunit using activating and inhibiting antibodies. J Biol Chem. 1993 Aug 15;268(23):17597–17601. [PubMed] [Google Scholar]
  41. Tanaka Y., Albelda S. M., Horgan K. J., van Seventer G. A., Shimizu Y., Newman W., Hallam J., Newman P. J., Buck C. A., Shaw S. CD31 expressed on distinctive T cell subsets is a preferential amplifier of beta 1 integrin-mediated adhesion. J Exp Med. 1992 Jul 1;176(1):245–253. doi: 10.1084/jem.176.1.245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tavassoli M., Friedenstein A. Hemopoietic stromal microenvironment. Am J Hematol. 1983 Sep;15(2):195–203. doi: 10.1002/ajh.2830150211. [DOI] [PubMed] [Google Scholar]
  43. Teixidó J., Hemler M. E., Greenberger J. S., Anklesaria P. Role of beta 1 and beta 2 integrins in the adhesion of human CD34hi stem cells to bone marrow stroma. J Clin Invest. 1992 Aug;90(2):358–367. doi: 10.1172/JCI115870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Torok-Storb B. Cellular interactions. Blood. 1988 Aug;72(2):373–385. [PubMed] [Google Scholar]
  45. Toullec D., Pianetti P., Coste H., Bellevergue P., Grand-Perret T., Ajakane M., Baudet V., Boissin P., Boursier E., Loriolle F. The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. J Biol Chem. 1991 Aug 25;266(24):15771–15781. [PubMed] [Google Scholar]
  46. Verfaillie C., Blakolmer K., McGlave P. Purified primitive human hematopoietic progenitor cells with long-term in vitro repopulating capacity adhere selectively to irradiated bone marrow stroma. J Exp Med. 1990 Aug 1;172(2):509–502. doi: 10.1084/jem.172.2.509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wayner E. A., Garcia-Pardo A., Humphries M. J., McDonald J. A., Carter W. G. Identification and characterization of the T lymphocyte adhesion receptor for an alternative cell attachment domain (CS-1) in plasma fibronectin. J Cell Biol. 1989 Sep;109(3):1321–1330. doi: 10.1083/jcb.109.3.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Weinstein R., Riordan M. A., Wenc K., Kreczko S., Zhou M., Dainiak N. Dual role of fibronectin in hematopoietic differentiation. Blood. 1989 Jan;73(1):111–116. [PubMed] [Google Scholar]
  49. Williams D. A., Rios M., Stephens C., Patel V. P. Fibronectin and VLA-4 in haematopoietic stem cell-microenvironment interactions. Nature. 1991 Aug 1;352(6334):438–441. doi: 10.1038/352438a0. [DOI] [PubMed] [Google Scholar]
  50. Williams M. J., Hughes P. E., O'Toole T. E., Ginsberg M. H. The inner world of cell adhesion: integrin cytoplasmic domains. Trends Cell Biol. 1994 Apr;4(4):109–112. doi: 10.1016/0962-8924(94)90059-0. [DOI] [PubMed] [Google Scholar]
  51. Yanai N., Sekine C., Yagita H., Obinata M. Roles for integrin very late activation antigen-4 in stroma-dependent erythropoiesis. Blood. 1994 May 15;83(10):2844–2850. [PubMed] [Google Scholar]
  52. Ylänne J., Chen Y., O'Toole T. E., Loftus J. C., Takada Y., Ginsberg M. H. Distinct functions of integrin alpha and beta subunit cytoplasmic domains in cell spreading and formation of focal adhesions. J Cell Biol. 1993 Jul;122(1):223–233. doi: 10.1083/jcb.122.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Zhou Y. Q., Levesque J. P., Hatzfeld A., Cardoso A. A., Li M. L., Sansilvestri P., Hatzfeld J. Fibrinogen potentiates the effect of interleukin-3 on early human hematopoietic progenitors. Blood. 1993 Aug 1;82(3):800–806. [PubMed] [Google Scholar]

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