Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1998 Jun 1;101(11):2456–2467. doi: 10.1172/JCI188

Mobilization and homing of peripheral blood progenitors is related to reversible downregulation of alpha4 beta1 integrin expression and function.

F Prosper 1, D Stroncek 1, J B McCarthy 1, C M Verfaillie 1
PMCID: PMC508835  PMID: 9616217

Abstract

Despite the wide use of mobilized peripheral blood (PB) progenitor cells (PBPC) for clinical transplantation the mechanism(s) underlying their mobilization and subsequent engraftment are still unknown. We compared the adhesive phenotype of CD34(+) colony-forming cells (CFC) in bone marrow (BM) and PB of normal donors before and after administration of granulocyte colony-stimulating factor (G-CSF) for 5 d. G-CSF-mobilized PB CFC cells adhered significantly less to BM stroma, fibronectin, and to the alpha4 beta1 binding fibronectin peptide, CS1, because of decreased expression of the alpha4 integrin. Since incubation of BM CD34(+) cells for 4 d with G-CSF at concentrations found in serum of G-CSF- treated individuals did not affect alpha4-dependent adhesion, G-CSF may not be directly responsible for the decreased alpha4-mediated adhesion of PB CFC. Culture of G-CSF-mobilized PB CD34(+) cells with cytokines at concentrations found in BM stromal cultures upregulated alpha4 expression and restored adhesion of mobilized PB CFC to stroma, fibronectin, and CS1. Adhesion of cultured, mobilized PB CFC to stroma and CS1 could not be further upregulated by the beta1 activating antibody, 8A2. This indicates acquisition of a maximally activated alpha4 beta1 integrin once PB CFC have been removed from the in vivo mobilizing milieu. Thus, decreased alpha4 expression on CD34(+) CFC in PB may be responsible for the aberrant circulation of mobilized PB CD34(+) cells. Reexpression of a maximally activated alpha4 beta1 integrin on mobilized PB CFC removed from the mobilizing in vivo milieu may contribute to the early engraftment of mobilized PBPC.

Full Text

The Full Text of this article is available as a PDF (255.3 KB).

Selected References

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

  1. Barrett A. J., Longhurst P., Sneath P., Watson J. G. Mobilization of CFU-C by exercise and ACTH induced stress in man. Exp Hematol. 1978 Aug;6(7):590–594. [PubMed] [Google Scholar]
  2. Bensinger W., Singer J., Appelbaum F., Lilleby K., Longin K., Rowley S., Clarke E., Clift R., Hansen J., Shields T. Autologous transplantation with peripheral blood mononuclear cells collected after administration of recombinant granulocyte stimulating factor. Blood. 1993 Jun 1;81(11):3158–3163. [PubMed] [Google Scholar]
  3. Bhatia R., McGlave P. B., Verfaillie C. M. Treatment of marrow stroma with interferon-alpha restores normal beta 1 integrin-dependent adhesion of chronic myelogenous leukemia hematopoietic progenitors. Role of MIP-1 alpha. J Clin Invest. 1995 Aug;96(2):931–939. doi: 10.1172/JCI118141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bhatia R., Wayner E. A., McGlave P. B., Verfaillie C. M. Interferon-alpha restores normal adhesion of chronic myelogenous leukemia hematopoietic progenitors to bone marrow stroma by correcting impaired beta 1 integrin receptor function. J Clin Invest. 1994 Jul;94(1):384–391. doi: 10.1172/JCI117333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. CAVINS J. A., SCHEER S. C., THOMAS E. D., FERREBEE J. W. THE RECOVERY OF LETHALLY IRRADIATED DOGS GIVEN INFUSIONS OF AUTOLOGOUS LEUKOCYTES PRESERVED AT -80 C. Blood. 1964 Jan;23:38–42. [PubMed] [Google Scholar]
  6. Chervenick P. A., Boggs D. R. In vitro growth of granulocytic and mononuclear cell colonies from blood of normal individuals. Blood. 1971 Feb;37(2):131–135. [PubMed] [Google Scholar]
  7. Clarke B. J., Housman D. Characterization of an erythroid precursor cell of high proliferative capacity in normal human peripheral blood. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1105–1109. doi: 10.1073/pnas.74.3.1105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cline M. J., Golde D. W. Mobilization of hematopoietic stem cells (CFU-C) into the peripheral blood of man by endotoxin. Exp Hematol. 1977 May;5(3):186–190. [PubMed] [Google Scholar]
  9. Coulombel L., Eaves A. C., Eaves C. J. Enzymatic treatment of long-term human marrow cultures reveals the preferential location of primitive hemopoietic progenitors in the adherent layer. Blood. 1983 Aug;62(2):291–297. [PubMed] [Google Scholar]
  10. Dercksen M. W., Gerritsen W. R., Rodenhuis S., Dirkson M. K., Slaper-Cortenbach I. C., Schaasberg W. P., Pinedo H. M., von dem Borne A. E., van der Schoot C. E. Expression of adhesion molecules on CD34+ cells: CD34+ L-selectin+ cells predict a rapid platelet recovery after peripheral blood stem cell transplantation. Blood. 1995 Jun 1;85(11):3313–3319. [PubMed] [Google Scholar]
  11. Donahue R. E., Kirby M. R., Metzger M. E., Agricola B. A., Sellers S. E., Cullis H. M. Peripheral blood CD34+ cells differ from bone marrow CD34+ cells in Thy-1 expression and cell cycle status in nonhuman primates mobilized or not mobilized with granulocyte colony-stimulating factor and/or stem cell factor. Blood. 1996 Feb 15;87(4):1644–1653. [PubMed] [Google Scholar]
  12. 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]
  13. Fuhlbrigge R. C., Alon R., Puri K. D., Lowe J. B., Springer T. A. Sialylated, fucosylated ligands for L-selectin expressed on leukocytes mediate tethering and rolling adhesions in physiologic flow conditions. J Cell Biol. 1996 Nov;135(3):837–848. doi: 10.1083/jcb.135.3.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fujisaki T., Otsuka T., Harada M., Ohno Y., Niho Y. Granulocyte colony-stimulating factor mobilizes primitive hematopoietic stem cells in normal individuals. Bone Marrow Transplant. 1995 Jul;16(1):57–62. [PubMed] [Google Scholar]
  15. GOODMAN J. W., HODGSON G. S. Evidence for stem cells in the peripheral blood of mice. Blood. 1962 Jun;19:702–714. [PubMed] [Google Scholar]
  16. Guan J. L., Hynes R. O. Lymphoid cells recognize an alternatively spliced segment of fibronectin via the integrin receptor alpha 4 beta 1. Cell. 1990 Jan 12;60(1):53–61. doi: 10.1016/0092-8674(90)90715-q. [DOI] [PubMed] [Google Scholar]
  17. Gupta P., McCarthy J. B., Verfaillie C. M. Stromal fibroblast heparan sulfate is required for cytokine-mediated ex vivo maintenance of human long-term culture-initiating cells. Blood. 1996 Apr 15;87(8):3229–3236. [PubMed] [Google Scholar]
  18. Hardy C. L. The homing of hematopoietic stem cells to the bone marrow. Am J Med Sci. 1995 May;309(5):260–266. doi: 10.1097/00000441-199530950-00005. [DOI] [PubMed] [Google Scholar]
  19. Huhtala P., Humphries M. J., McCarthy J. B., Tremble P. M., Werb Z., Damsky C. H. Cooperative signaling by alpha 5 beta 1 and alpha 4 beta 1 integrins regulates metalloproteinase gene expression in fibroblasts adhering to fibronectin. J Cell Biol. 1995 May;129(3):867–879. doi: 10.1083/jcb.129.3.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jalkanen S., Jalkanen M., Bargatze R., Tammi M., Butcher E. C. Biochemical properties of glycoproteins involved in lymphocyte recognition of high endothelial venules in man. J Immunol. 1988 Sep 1;141(5):1615–1623. [PubMed] [Google Scholar]
  21. Kassner P. D., Hemler M. E. Interchangeable alpha chain cytoplasmic domains play a positive role in control of cell adhesion mediated by VLA-4, a beta 1 integrin. J Exp Med. 1993 Aug 1;178(2):649–660. doi: 10.1084/jem.178.2.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Kessinger A., Armitage J. O., Landmark J. D., Weisenburger D. D. Reconstitution of human hematopoietic function with autologous cryopreserved circulating stem cells. Exp Hematol. 1986 Mar;14(3):192–196. [PubMed] [Google Scholar]
  24. Kinashi T., Springer T. A. Adhesion molecules in hematopoietic cells. Blood Cells. 1994;20(1):25–44. [PubMed] [Google Scholar]
  25. Kodama H., Nose M., Niida S., Nishikawa S., Nishikawa S. Involvement of the c-kit receptor in the adhesion of hematopoietic stem cells to stromal cells. Exp Hematol. 1994 Sep;22(10):979–984. [PubMed] [Google Scholar]
  26. 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]
  27. Kovach N. L., Lin N., Yednock T., Harlan J. M., Broudy V. C. Stem cell factor modulates avidity of alpha 4 beta 1 and alpha 5 beta 1 integrins expressed on hematopoietic cell lines. Blood. 1995 Jan 1;85(1):159–167. [PubMed] [Google Scholar]
  28. Lane T. A., Law P., Maruyama M., Young D., Burgess J., Mullen M., Mealiffe M., Terstappen L. W., Hardwick A., Moubayed M. Harvesting and enrichment of hematopoietic progenitor cells mobilized into the peripheral blood of normal donors by granulocyte-macrophage colony-stimulating factor (GM-CSF) or G-CSF: potential role in allogeneic marrow transplantation. Blood. 1995 Jan 1;85(1):275–282. [PubMed] [Google Scholar]
  29. Laszik Z., Jansen P. J., Cummings R. D., Tedder T. F., McEver R. P., Moore K. L. P-selectin glycoprotein ligand-1 is broadly expressed in cells of myeloid, lymphoid, and dendritic lineage and in some nonhematopoietic cells. Blood. 1996 Oct 15;88(8):3010–3021. [PubMed] [Google Scholar]
  30. 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]
  31. Lundell B. I., McCarthy J. B., Kovach N. L., Verfaillie C. M. Activation-dependent alpha5beta1 integrin-mediated adhesion to fibronectin decreases proliferation of chronic myelogenous leukemia progenitors and K562 cells. Blood. 1996 Mar 15;87(6):2450–2458. [PubMed] [Google Scholar]
  32. Lévesque J. P., Leavesley D. I., Niutta S., Vadas M., Simmons P. J. Cytokines increase human hemopoietic cell adhesiveness by activation of very late antigen (VLA)-4 and VLA-5 integrins. J Exp Med. 1995 May 1;181(5):1805–1815. doi: 10.1084/jem.181.5.1805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. McCarthy J. B., Hagen S. T., Furcht L. T. Human fibronectin contains distinct adhesion- and motility-promoting domains for metastatic melanoma cells. J Cell Biol. 1986 Jan;102(1):179–188. doi: 10.1083/jcb.102.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. McCarthy J. B., Skubitz A. P., Qi Z., Yi X. Y., Mickelson D. J., Klein D. J., Furcht L. T. RGD-independent cell adhesion to the carboxy-terminal heparin-binding fragment of fibronectin involves heparin-dependent and -independent activities. J Cell Biol. 1990 Mar;110(3):777–787. doi: 10.1083/jcb.110.3.777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. McCredie K. B., Hersh E. M., Freireich E. J. Cells capable of colony formation in the peripheral blood of man. Science. 1971 Jan 22;171(3968):293–294. doi: 10.1126/science.171.3968.293. [DOI] [PubMed] [Google Scholar]
  36. Möhle R., Murea S., Kirsch M., Haas R. Differential expression of L-selectin, VLA-4, and LFA-1 on CD34+ progenitor cells from bone marrow and peripheral blood during G-CSF-enhanced recovery. Exp Hematol. 1995 Dec;23(14):1535–1542. [PubMed] [Google Scholar]
  37. Papayannopoulou T., Craddock C. Homing and trafficking of hemopoietic progenitor cells. Acta Haematol. 1997;97(1-2):97–104. doi: 10.1159/000203665. [DOI] [PubMed] [Google Scholar]
  38. Papayannopoulou T., Craddock C., Nakamoto B., Priestley G. V., Wolf N. S. The VLA4/VCAM-1 adhesion pathway defines contrasting mechanisms of lodgement of transplanted murine hemopoietic progenitors between bone marrow and spleen. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9647–9651. doi: 10.1073/pnas.92.21.9647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Papayannopoulou T., Nakamoto B. Peripheralization of hemopoietic progenitors in primates treated with anti-VLA4 integrin. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9374–9378. doi: 10.1073/pnas.90.20.9374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Petzer A. L., Hogge D. E., Landsdorp P. M., Reid D. S., Eaves C. J. Self-renewal of primitive human hematopoietic cells (long-term-culture-initiating cells) in vitro and their expansion in defined medium. Proc Natl Acad Sci U S A. 1996 Feb 20;93(4):1470–1474. doi: 10.1073/pnas.93.4.1470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Prosper F., Stroncek D., Verfaillie C. M. Phenotypic and functional characterization of long-term culture-initiating cells present in peripheral blood progenitor collections of normal donors treated with granulocyte colony-stimulating factor. Blood. 1996 Sep 15;88(6):2033–2042. [PubMed] [Google Scholar]
  42. Rice G. E., Munro J. M., Bevilacqua M. P. Inducible cell adhesion molecule 110 (INCAM-110) is an endothelial receptor for lymphocytes. A CD11/CD18-independent adhesion mechanism. J Exp Med. 1990 Apr 1;171(4):1369–1374. doi: 10.1084/jem.171.4.1369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sheridan W. P., Begley C. G., Juttner C. A., Szer J., To L. B., Maher D., McGrath K. M., Morstyn G., Fox R. M. Effect of peripheral-blood progenitor cells mobilised by filgrastim (G-CSF) on platelet recovery after high-dose chemotherapy. Lancet. 1992 Mar 14;339(8794):640–644. doi: 10.1016/0140-6736(92)90795-5. [DOI] [PubMed] [Google Scholar]
  44. Siena S., Bregni M., Brando B., Ravagnani F., Bonadonna G., Gianni A. M. Circulation of CD34+ hematopoietic stem cells in the peripheral blood of high-dose cyclophosphamide-treated patients: enhancement by intravenous recombinant human granulocyte-macrophage colony-stimulating factor. Blood. 1989 Nov 1;74(6):1905–1914. [PubMed] [Google Scholar]
  45. 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]
  46. Simmons P. J., Masinovsky B., Longenecker B. M., Berenson R., Torok-Storb B., Gallatin W. M. Vascular cell adhesion molecule-1 expressed by bone marrow stromal cells mediates the binding of hematopoietic progenitor cells. Blood. 1992 Jul 15;80(2):388–395. [PubMed] [Google Scholar]
  47. 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]
  48. Simon S. I., Burns A. R., Taylor A. D., Gopalan P. K., Lynam E. B., Sklar L. A., Smith C. W. L-selectin (CD62L) cross-linking signals neutrophil adhesive functions via the Mac-1 (CD11b/CD18) beta 2-integrin. J Immunol. 1995 Aug 1;155(3):1502–1514. [PubMed] [Google Scholar]
  49. Stute N., Santana V. M., Rodman J. H., Schell M. J., Ihle J. N., Evans W. E. Pharmacokinetics of subcutaneous recombinant human granulocyte colony-stimulating factor in children. Blood. 1992 Jun 1;79(11):2849–2854. [PubMed] [Google Scholar]
  50. Sutherland D. R., Keating A., Nayar R., Anania S., Stewart A. K. Sensitive detection and enumeration of CD34+ cells in peripheral and cord blood by flow cytometry. Exp Hematol. 1994 Sep;22(10):1003–1010. [PubMed] [Google Scholar]
  51. Taswell C. Limiting dilution assays for the determination of immunocompetent cell frequencies. I. Data analysis. J Immunol. 1981 Apr;126(4):1614–1619. [PubMed] [Google Scholar]
  52. Tavassoli M. Expansion of blood stem cell pool or mobilization of its marrow counterpart? Exp Hematol. 1993 Aug;21(9):1205–1206. [PubMed] [Google Scholar]
  53. 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]
  54. Teofili L., Iovino M. S., Sica S., Pierelli L., Menichella G., De Stefano V., Rumi C., Leone G. Characterization of peripheral blood CD34+ progenitor cells mobilized with chemotherapy and granulocyte colony-stimulating factor. Exp Hematol. 1994 Sep;22(10):990–995. [PubMed] [Google Scholar]
  55. Turner M. L., McIlwaine K., Anthony R. S., Parker A. C. Differential expression of cell adhesion molecules by human hematopoietic progenitor cells from bone marrow and mobilized adult peripheral blood. Stem Cells. 1995 May;13(3):311–316. doi: 10.1002/stem.5530130312. [DOI] [PubMed] [Google Scholar]
  56. Verfaillie C. M. Can human hematopoietic stem cells be cultured ex vivo? Stem Cells. 1994 Sep;12(5):466–476. doi: 10.1002/stem.5530120503. [DOI] [PubMed] [Google Scholar]
  57. Verfaillie C. M. Direct contact between human primitive hematopoietic progenitors and bone marrow stroma is not required for long-term in vitro hematopoiesis. Blood. 1992 Jun 1;79(11):2821–2826. [PubMed] [Google Scholar]
  58. Verfaillie C. M., McCarthy J. B., McGlave P. B. Differentiation of primitive human multipotent hematopoietic progenitors into single lineage clonogenic progenitors is accompanied by alterations in their interaction with fibronectin. J Exp Med. 1991 Sep 1;174(3):693–703. doi: 10.1084/jem.174.3.693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. 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]
  60. Watt S. M., Williamson J., Genevier H., Fawcett J., Simmons D. L., Hatzfeld A., Nesbitt S. A., Coombe D. R. The heparin binding PECAM-1 adhesion molecule is expressed by CD34+ hematopoietic precursor cells with early myeloid and B-lymphoid cell phenotypes. Blood. 1993 Nov 1;82(9):2649–2663. [PubMed] [Google Scholar]
  61. 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]
  62. Zannettino A. C., Berndt M. C., Butcher C., Butcher E. C., Vadas M. A., Simmons P. J. Primitive human hematopoietic progenitors adhere to P-selectin (CD62P). Blood. 1995 Jun 15;85(12):3466–3477. [PubMed] [Google Scholar]

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

RESOURCES