Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1995 Mar 28;92(7):2859–2863. doi: 10.1073/pnas.92.7.2859

gp130 and c-Kit signalings synergize for ex vivo expansion of human primitive hemopoietic progenitor cells.

X Sui 1, K Tsuji 1, R Tanaka 1, S Tajima 1, K Muraoka 1, Y Ebihara 1, K Ikebuchi 1, K Yasukawa 1, T Taga 1, T Kishimoto 1, et al.
PMCID: PMC42318  PMID: 7535932

Abstract

gp130, a signal-transducing receptor component of interleukin 6 (IL-6), associates with an IL-6 and IL-6 receptor (IL-6) complex and transduces signals. To examine the role of gp130 signaling in the expansion of human hemopoietic progenitor cells, we tested the effects of a recombinant soluble human IL-6 receptor (sIL-6R) and/or IL-6 in combination with other cytokines on purified human umbilical cord blood CD34+ cells, using methylcellulose clonal assay and suspension culture in the presence or absence of serum. A combination of sIL-6R and IL-6 (sIL-6R/IL-6), but not sIL-6R or IL-6 alone, was found to dramatically stimulate expansion of hemopoietic progenitor cells as well as CD34+ cells in the presence of stem cell factor. Significant generation of multipotential hemopoietic progenitors over a period of 3 weeks in suspension culture and efficient formation of colonies, especially multilineage and blast cell colonies, in methylcellulose assay supplemented with a combination of sIL-6R/IL-6 together with stem cell factor were observed in serum-containing and serum-free culture. Addition of anti-gp130 monoclonal antibodies or anti-IL-6R monoclonal antibodies to the above cultures dose-dependently inhibited the expansion of progenitor cells in suspension culture and also completely blocked the formation of multilineage colonies in methylcellulose culture. These findings demonstrated that the significant expansion of human primitive hemopoietic progenitors could be achieved with the gp130 and c-Kit signalings initiated by the sIL-6R/IL-6 complex in the presence of stem cell factor and suggested the possible application of this method for ex vivo expansion of CD34+ cells for bone marrow transplantation.

Full text

PDF
2859

Selected References

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

  1. Brugger W., Möcklin W., Heimfeld S., Berenson R. J., Mertelsmann R., Kanz L. Ex vivo expansion of enriched peripheral blood CD34+ progenitor cells by stem cell factor, interleukin-1 beta (IL-1 beta), IL-6, IL-3, interferon-gamma, and erythropoietin. Blood. 1993 May 15;81(10):2579–2584. [PubMed] [Google Scholar]
  2. Haylock D. N., To L. B., Dowse T. L., Juttner C. A., Simmons P. J. Ex vivo expansion and maturation of peripheral blood CD34+ cells into the myeloid lineage. Blood. 1992 Sep 15;80(6):1405–1412. [PubMed] [Google Scholar]
  3. Hibi M., Murakami M., Saito M., Hirano T., Taga T., Kishimoto T. Molecular cloning and expression of an IL-6 signal transducer, gp130. Cell. 1990 Dec 21;63(6):1149–1157. doi: 10.1016/0092-8674(90)90411-7. [DOI] [PubMed] [Google Scholar]
  4. Hirata Y., Taga T., Hibi M., Nakano N., Hirano T., Kishimoto T. Characterization of IL-6 receptor expression by monoclonal and polyclonal antibodies. J Immunol. 1989 Nov 1;143(9):2900–2906. [PubMed] [Google Scholar]
  5. Ikebuchi K., Wong G. G., Clark S. C., Ihle J. N., Hirai Y., Ogawa M. Interleukin 6 enhancement of interleukin 3-dependent proliferation of multipotential hemopoietic progenitors. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9035–9039. doi: 10.1073/pnas.84.24.9035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Imai T., Koike K., Kubo T., Kikuchi T., Amano Y., Takagi M., Okumura N., Nakahata T. Interleukin-6 supports human megakaryocytic proliferation and differentiation in vitro. Blood. 1991 Oct 15;78(8):1969–1974. [PubMed] [Google Scholar]
  7. Iscove N. N., Shaw A. R., Keller G. Net increase of pluripotential hematopoietic precursors in suspension culture in response to IL-1 and IL-3. J Immunol. 1989 Apr 1;142(7):2332–2337. [PubMed] [Google Scholar]
  8. Kishimoto T., Akira S., Taga T. Interleukin-6 and its receptor: a paradigm for cytokines. Science. 1992 Oct 23;258(5082):593–597. doi: 10.1126/science.1411569. [DOI] [PubMed] [Google Scholar]
  9. Kishimoto T., Taga T., Akira S. Cytokine signal transduction. Cell. 1994 Jan 28;76(2):253–262. doi: 10.1016/0092-8674(94)90333-6. [DOI] [PubMed] [Google Scholar]
  10. Koike K., Nakahata T., Takagi M., Kobayashi T., Ishiguro A., Tsuji K., Naganuma K., Okano A., Akiyama Y., Akabane T. Synergism of BSF-2/interleukin 6 and interleukin 3 on development of multipotential hemopoietic progenitors in serum-free culture. J Exp Med. 1988 Sep 1;168(3):879–890. doi: 10.1084/jem.168.3.879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Koller M. R., Emerson S. G., Palsson B. O. Large-scale expansion of human stem and progenitor cells from bone marrow mononuclear cells in continuous perfusion cultures. Blood. 1993 Jul 15;82(2):378–384. [PubMed] [Google Scholar]
  12. Mackiewicz A., Schooltink H., Heinrich P. C., Rose-John S. Complex of soluble human IL-6-receptor/IL-6 up-regulates expression of acute-phase proteins. J Immunol. 1992 Sep 15;149(6):2021–2027. [PubMed] [Google Scholar]
  13. Mayani H., Lansdorp P. M. Thy-1 expression is linked to functional properties of primitive hematopoietic progenitor cells from human umbilical cord blood. Blood. 1994 May 1;83(9):2410–2417. [PubMed] [Google Scholar]
  14. Murakami M., Hibi M., Nakagawa N., Nakagawa T., Yasukawa K., Yamanishi K., Taga T., Kishimoto T. IL-6-induced homodimerization of gp130 and associated activation of a tyrosine kinase. Science. 1993 Jun 18;260(5115):1808–1810. doi: 10.1126/science.8511589. [DOI] [PubMed] [Google Scholar]
  15. Nakahata T., Ogawa M. Hemopoietic colony-forming cells in umbilical cord blood with extensive capability to generate mono- and multipotential hemopoietic progenitors. J Clin Invest. 1982 Dec;70(6):1324–1328. doi: 10.1172/JCI110734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Saito M., Yoshida K., Hibi M., Taga T., Kishimoto T. Molecular cloning of a murine IL-6 receptor-associated signal transducer, gp130, and its regulated expression in vivo. J Immunol. 1992 Jun 15;148(12):4066–4071. [PubMed] [Google Scholar]
  17. Saito T., Taga T., Miki D., Futatsugi K., Yawata H., Kishimoto T., Yasukawa K. Preparation of monoclonal antibodies against the IL-6 signal transducer, gp130, that can inhibit IL-6-mediated functions. J Immunol Methods. 1993 Aug 9;163(2):217–223. doi: 10.1016/0022-1759(93)90125-q. [DOI] [PubMed] [Google Scholar]
  18. Sato N., Sawada K., Koizumi K., Tarumi T., Ieko M., Yasukouchi T., Yamaguchi M., Takahashi T. A., Sekiguchi S., Koike T. In vitro expansion of human peripheral blood CD34+ cells. Blood. 1993 Dec 15;82(12):3600–3609. [PubMed] [Google Scholar]
  19. Taga T., Hibi M., Hirata Y., Yamasaki K., Yasukawa K., Matsuda T., Hirano T., Kishimoto T. Interleukin-6 triggers the association of its receptor with a possible signal transducer, gp130. Cell. 1989 Aug 11;58(3):573–581. doi: 10.1016/0092-8674(89)90438-8. [DOI] [PubMed] [Google Scholar]
  20. Taga T., Narazaki M., Yasukawa K., Saito T., Miki D., Hamaguchi M., Davis S., Shoyab M., Yancopoulos G. D., Kishimoto T. Functional inhibition of hematopoietic and neurotrophic cytokines by blocking the interleukin 6 signal transducer gp130. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10998–11001. doi: 10.1073/pnas.89.22.10998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tamura T., Udagawa N., Takahashi N., Miyaura C., Tanaka S., Yamada Y., Koishihara Y., Ohsugi Y., Kumaki K., Taga T. Soluble interleukin-6 receptor triggers osteoclast formation by interleukin 6. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11924–11928. doi: 10.1073/pnas.90.24.11924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tanaka R., Koike K., Imai T., Shiohara M., Kubo T., Amano Y., Komiyama A., Nakahata T. Stem cell factor enhances proliferation, but not maturation, of murine megakaryocytic progenitors in serum-free culture. Blood. 1992 Oct 1;80(7):1743–1749. [PubMed] [Google Scholar]
  23. Yasukawa K., Futatsugi K., Saito T., Yawata H., Narazaki M., Suzuki H., Taga T., Kishimoto T. Association of recombinant soluble IL-6-signal transducer, gp130, with a complex of IL 6 and soluble IL-6 receptor, and establishment of an ELISA for soluble gp130. Immunol Lett. 1992 Feb;31(2):123–130. doi: 10.1016/0165-2478(92)90138-e. [DOI] [PubMed] [Google Scholar]
  24. Yasukawa K., Saito T., Fukunaga T., Sekimori Y., Koishihara Y., Fukui H., Ohsugi Y., Matsuda T., Yawata H., Hirano T. Purification and characterization of soluble human IL-6 receptor expressed in CHO cells. J Biochem. 1990 Oct;108(4):673–676. doi: 10.1093/oxfordjournals.jbchem.a123261. [DOI] [PubMed] [Google Scholar]
  25. Yoshida K., Chambers I., Nichols J., Smith A., Saito M., Yasukawa K., Shoyab M., Taga T., Kishimoto T. Maintenance of the pluripotential phenotype of embryonic stem cells through direct activation of gp130 signalling pathways. Mech Dev. 1994 Feb;45(2):163–171. doi: 10.1016/0925-4773(94)90030-2. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES