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. 1993 Sep 1;178(3):925–932. doi: 10.1084/jem.178.3.925

Transforming growth factor beta: is it a downregulator of stem cell inhibition by macrophage inflammatory protein 1 alpha?

PMCID: PMC2191149  PMID: 8394405

Abstract

Transforming growth factor beta 1 (TGF-beta 1) and macrophage inflammatory protein 1 alpha (MIP-1 alpha) have recently been identified as potent inhibitors of hemopoietic stem cell proliferation. From previous studies, these molecules appear to have similar functions in the control of stem cell proliferation. This study was designed to investigate the relationship, if any, between these two negative regulators in an attempt to elucidate possible distinctive roles for each within the hemopoietic system. We report here that both MIP-1 alpha and TGF-beta are capable of inhibiting the same stem cell population (colony-forming unit [CFU]-A/CFU-S) with similar potencies. We further show that TGF-beta potently inhibits MIP-1 alpha gene expression in bone marrow-derived macrophages, the presumed source of MIP-1 alpha in the bone marrow. This inhibition is not specific to MIP- 1 alpha in that expression of MIP-1 beta, a related molecule that does not exhibit potent stem cell inhibitory properties, is inhibited in a similar manner. The inhibition of MIP-1 alpha gene expression is also seen as a reduction in MIP-1 alpha protein production, which markedly decreases 24 h after treating RAW 264.7 cells, a murine macrophage cell line, with TGF-beta. These in vitro results suggest that in the presence of active TGF-beta in vivo, and in the absence of upregulators of MIP-1 alpha transcription, very little MIP-1 alpha will be produced. To address how MIP-1 alpha's target cells, the stem cells, would respond to TGF-beta, and the consequently low levels of MIP-1 alpha produced, we analyzed the effect of TGF-beta on MIP-1 alpha receptor levels on FDCP-MIX cells, a murine stem cell line. We show that TGF- beta (100 pM) reversibly downregulates MIP-1 alpha receptor levels on these cells to a maximum of 50-70% after 24 h. This level of downregulation does not change upon increasing the concentration of TGF- beta or the length of exposure of the cells to TGF-beta. Scatchard analysis shows that TGF-beta downregulates MIP-1 alpha receptor numbers with no change in affinity of the remaining receptors. These results suggest that TGF-beta may be capable of interfering with MIP-1 alpha's role as a stem cell inhibitor. Indeed, they suggest that in the presence of active TGF-beta in vivo, MIP-1 alpha is at best a weak contributor to the overall physiological inhibition of stem cells.

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Selected References

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  1. Akhurst R. J., Fee F., Balmain A. Localized production of TGF-beta mRNA in tumour promoter-stimulated mouse epidermis. Nature. 1988 Jan 28;331(6154):363–365. doi: 10.1038/331363a0. [DOI] [PubMed] [Google Scholar]
  2. Bonewald L. F. Can transforming growth factor beta be useful as a protective agent for pluripotent hematopoietic progenitor cells? Exp Hematol. 1992 Dec;20(11):1249–1251. [PubMed] [Google Scholar]
  3. Broxmeyer H. E., Sherry B., Lu L., Cooper S., Oh K. O., Tekamp-Olson P., Kwon B. S., Cerami A. Enhancing and suppressing effects of recombinant murine macrophage inflammatory proteins on colony formation in vitro by bone marrow myeloid progenitor cells. Blood. 1990 Sep 15;76(6):1110–1116. [PubMed] [Google Scholar]
  4. Chantry D., Turner M., Abney E., Feldmann M. Modulation of cytokine production by transforming growth factor-beta. J Immunol. 1989 Jun 15;142(12):4295–4300. [PubMed] [Google Scholar]
  5. Clements J. M., Craig S., Gearing A. J., Hunter M. G., Heyworth C. M., Dexter T. M., Lord B. I. Biological and structural properties of MIP-1 alpha expressed in yeast. Cytokine. 1992 Jan;4(1):76–82. doi: 10.1016/1043-4666(92)90040-x. [DOI] [PubMed] [Google Scholar]
  6. Dunlop D. J., Wright E. G., Lorimore S., Graham G. J., Holyoake T., Kerr D. J., Wolpe S. D., Pragnell I. B. Demonstration of stem cell inhibition and myeloprotective effects of SCI/rhMIP1 alpha in vivo. Blood. 1992 May 1;79(9):2221–2225. [PubMed] [Google Scholar]
  7. Gao J. L., Kuhns D. B., Tiffany H. L., McDermott D., Li X., Francke U., Murphy P. M. Structure and functional expression of the human macrophage inflammatory protein 1 alpha/RANTES receptor. J Exp Med. 1993 May 1;177(5):1421–1427. doi: 10.1084/jem.177.5.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Graham G. J., Freshney M. G., Donaldson D., Pragnell I. B. Purification and biochemical characterisation of human and murine stem cell inhibitors (SCI). Growth Factors. 1992;7(2):151–160. doi: 10.3109/08977199209046404. [DOI] [PubMed] [Google Scholar]
  9. Graham G. J., Pragnell I. B. Negative regulators of haemopoiesis--current advances. Prog Growth Factor Res. 1990;2(3):181–192. doi: 10.1016/0955-2235(90)90004-4. [DOI] [PubMed] [Google Scholar]
  10. Graham G. J., Wright E. G., Hewick R., Wolpe S. D., Wilkie N. M., Donaldson D., Lorimore S., Pragnell I. B. Identification and characterization of an inhibitor of haemopoietic stem cell proliferation. Nature. 1990 Mar 29;344(6265):442–444. doi: 10.1038/344442a0. [DOI] [PubMed] [Google Scholar]
  11. Graham G. J., Zhou L., Weatherbee J. A., Tsang M. L., Napolitano M., Leonard W. J., Pragnell I. B. Characterization of a receptor for macrophage inflammatory protein 1 alpha and related proteins on human and murine cells. Cell Growth Differ. 1993 Mar;4(3):137–146. [PubMed] [Google Scholar]
  12. Heufler C., Topar G., Koch F., Trockenbacher B., Kämpgen E., Romani N., Schuler G. Cytokine gene expression in murine epidermal cell suspensions: interleukin 1 beta and macrophage inflammatory protein 1 alpha are selectively expressed in Langerhans cells but are differentially regulated in culture. J Exp Med. 1992 Oct 1;176(4):1221–1226. doi: 10.1084/jem.176.4.1221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Holmes W. E., Lee J., Kuang W. J., Rice G. C., Wood W. I. Structure and functional expression of a human interleukin-8 receptor. Science. 1991 Sep 13;253(5025):1278–1280. doi: 10.1126/science.1840701. [DOI] [PubMed] [Google Scholar]
  14. Jacobsen S. E., Keller J. R., Ruscetti F. W., Kondaiah P., Roberts A. B., Falk L. A. Bidirectional effects of transforming growth factor beta (TGF-beta) on colony-stimulating factor-induced human myelopoiesis in vitro: differential effects of distinct TGF-beta isoforms. Blood. 1991 Nov 1;78(9):2239–2247. [PubMed] [Google Scholar]
  15. Jacobsen S. E., Ruscetti F. W., Dubois C. M., Lee J., Boone T. C., Keller J. R. Transforming growth factor-beta trans-modulates the expression of colony stimulating factor receptors on murine hematopoietic progenitor cell lines. Blood. 1991 Apr 15;77(8):1706–1716. [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Laiho M., DeCaprio J. A., Ludlow J. W., Livingston D. M., Massagué J. Growth inhibition by TGF-beta linked to suppression of retinoblastoma protein phosphorylation. Cell. 1990 Jul 13;62(1):175–185. doi: 10.1016/0092-8674(90)90251-9. [DOI] [PubMed] [Google Scholar]
  18. Lenfant M., Wdzieczak-Bakala J., Guittet E., Prome J. C., Sotty D., Frindel E. Inhibitor of hematopoietic pluripotent stem cell proliferation: purification and determination of its structure. Proc Natl Acad Sci U S A. 1989 Feb;86(3):779–782. doi: 10.1073/pnas.86.3.779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lin H. Y., Wang X. F., Ng-Eaton E., Weinberg R. A., Lodish H. F. Expression cloning of the TGF-beta type II receptor, a functional transmembrane serine/threonine kinase. Cell. 1992 Feb 21;68(4):775–785. doi: 10.1016/0092-8674(92)90152-3. [DOI] [PubMed] [Google Scholar]
  20. Lord B. I., Dexter T. M., Clements J. M., Hunter M. A., Gearing A. J. Macrophage-inflammatory protein protects multipotent hematopoietic cells from the cytotoxic effects of hydroxyurea in vivo. Blood. 1992 May 15;79(10):2605–2609. [PubMed] [Google Scholar]
  21. Lorimore S. A., Pragnell I. B., Eckmann L., Wright E. G. Synergistic interactions allow colony formation in vitro by murine haemopoietic stem cells. Leuk Res. 1990;14(5):481–489. doi: 10.1016/0145-2126(90)90036-9. [DOI] [PubMed] [Google Scholar]
  22. Massagué J. Receptors for the TGF-beta family. Cell. 1992 Jun 26;69(7):1067–1070. doi: 10.1016/0092-8674(92)90627-o. [DOI] [PubMed] [Google Scholar]
  23. Massagué J. The transforming growth factor-beta family. Annu Rev Cell Biol. 1990;6:597–641. doi: 10.1146/annurev.cb.06.110190.003121. [DOI] [PubMed] [Google Scholar]
  24. McCartney-Francis N., Mizel D., Wong H., Wahl L., Wahl S. TGF-beta regulates production of growth factors and TGF-beta by human peripheral blood monocytes. Growth Factors. 1990;4(1):27–35. doi: 10.3109/08977199009011007. [DOI] [PubMed] [Google Scholar]
  25. Migdalska A., Molineux G., Demuynck H., Evans G. S., Ruscetti F., Dexter T. M. Growth inhibitory effects of transforming growth factor-beta 1 in vivo. Growth Factors. 1991;4(3):239–245. doi: 10.3109/08977199109104820. [DOI] [PubMed] [Google Scholar]
  26. Munson P. J., Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. doi: 10.1016/0003-2697(80)90515-1. [DOI] [PubMed] [Google Scholar]
  27. Murphy P. M., Tiffany H. L. Cloning of complementary DNA encoding a functional human interleukin-8 receptor. Science. 1991 Sep 13;253(5025):1280–1283. doi: 10.1126/science.1891716. [DOI] [PubMed] [Google Scholar]
  28. Neote K., DiGregorio D., Mak J. Y., Horuk R., Schall T. J. Molecular cloning, functional expression, and signaling characteristics of a C-C chemokine receptor. Cell. 1993 Feb 12;72(3):415–425. doi: 10.1016/0092-8674(93)90118-a. [DOI] [PubMed] [Google Scholar]
  29. Oh K. O., Zhou Z., Kim K. K., Samanta H., Fraser M., Kim Y. J., Broxmeyer H. E., Kwon B. S. Identification of cell surface receptors for murine macrophage inflammatory protein-1 alpha. J Immunol. 1991 Nov 1;147(9):2978–2983. [PubMed] [Google Scholar]
  30. Ohta M., Greenberger J. S., Anklesaria P., Bassols A., Massagué J. Two forms of transforming growth factor-beta distinguished by multipotential haematopoietic progenitor cells. Nature. 1987 Oct 8;329(6139):539–541. doi: 10.1038/329539a0. [DOI] [PubMed] [Google Scholar]
  31. Parkinson K., Balmain A. Chalones revisited--a possible role for transforming growth factor beta in tumour promotion. Carcinogenesis. 1990 Feb;11(2):195–198. doi: 10.1093/carcin/11.2.195. [DOI] [PubMed] [Google Scholar]
  32. Pietenpol J. A., Stein R. W., Moran E., Yaciuk P., Schlegel R., Lyons R. M., Pittelkow M. R., Münger K., Howley P. M., Moses H. L. TGF-beta 1 inhibition of c-myc transcription and growth in keratinocytes is abrogated by viral transforming proteins with pRB binding domains. Cell. 1990 Jun 1;61(5):777–785. doi: 10.1016/0092-8674(90)90188-k. [DOI] [PubMed] [Google Scholar]
  33. Pragnell I. B., Wright E. G., Lorimore S. A., Adam J., Rosendaal M., DeLamarter J. F., Freshney M., Eckmann L., Sproul A., Wilkie N. The effect of stem cell proliferation regulators demonstrated with an in vitro assay. Blood. 1988 Jul;72(1):196–201. [PubMed] [Google Scholar]
  34. Schall T. J. Biology of the RANTES/SIS cytokine family. Cytokine. 1991 May;3(3):165–183. doi: 10.1016/1043-4666(91)90013-4. [DOI] [PubMed] [Google Scholar]
  35. Shull M. M., Ormsby I., Kier A. B., Pawlowski S., Diebold R. J., Yin M., Allen R., Sidman C., Proetzel G., Calvin D. Targeted disruption of the mouse transforming growth factor-beta 1 gene results in multifocal inflammatory disease. Nature. 1992 Oct 22;359(6397):693–699. doi: 10.1038/359693a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Thompson N. L., Flanders K. C., Smith J. M., Ellingsworth L. R., Roberts A. B., Sporn M. B. Expression of transforming growth factor-beta 1 in specific cells and tissues of adult and neonatal mice. J Cell Biol. 1989 Feb;108(2):661–669. doi: 10.1083/jcb.108.2.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tsunawaki S., Sporn M., Ding A., Nathan C. Deactivation of macrophages by transforming growth factor-beta. Nature. 1988 Jul 21;334(6179):260–262. doi: 10.1038/334260a0. [DOI] [PubMed] [Google Scholar]
  38. Tushinski R. J., Stanley E. R. The regulation of mononuclear phagocyte entry into S phase by the colony stimulating factor CSF-1. J Cell Physiol. 1985 Feb;122(2):221–228. doi: 10.1002/jcp.1041220210. [DOI] [PubMed] [Google Scholar]
  39. Wang X. F., Lin H. Y., Ng-Eaton E., Downward J., Lodish H. F., Weinberg R. A. Expression cloning and characterization of the TGF-beta type III receptor. Cell. 1991 Nov 15;67(4):797–805. doi: 10.1016/0092-8674(91)90074-9. [DOI] [PubMed] [Google Scholar]
  40. Wrana J. L., Attisano L., Cárcamo J., Zentella A., Doody J., Laiho M., Wang X. F., Massagué J. TGF beta signals through a heteromeric protein kinase receptor complex. Cell. 1992 Dec 11;71(6):1003–1014. doi: 10.1016/0092-8674(92)90395-s. [DOI] [PubMed] [Google Scholar]
  41. Yan Z., Hsu S., Winawer S., Friedman E. Transforming growth factor beta 1 (TGF-beta 1) inhibits retinoblastoma gene expression but not pRB phosphorylation in TGF-beta 1-growth stimulated colon carcinoma cells. Oncogene. 1992 Apr;7(4):801–805. [PubMed] [Google Scholar]

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