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British Journal of Cancer logoLink to British Journal of Cancer
. 1983 Aug;48(2):247–259. doi: 10.1038/bjc.1983.180

Studies on the in vivo production of a lymphokine activity, interleukin 3 (IL-3) elaborated by lymphocytes and a myeloid leukaemic line in vitro and the fate of IL-3 dependent cell lines.

J M Garland, A Aldridge, J Wagstaffe, T M Dexter
PMCID: PMC2011446  PMID: 6411109

Abstract

Interleukin 3 (IL-3) is produced constitutively by WEHI-3b leukaemic cells and stimulated lymphoid cell populations in vitro. We have investigated the in vivo production of IL-3 in mice rendered leukaemic with WEHI-3b cells and mice stimulated by acute graft versus host disease (GVHD). In leukaemic mice, IL-3 was not found in serum or sonicates of 18-day spleens or bone marrow, although cells from the leukaemic organs were fully competent to elaborate IL-3 in vitro. Further, elaboration of IL-3 by WEHI cells in vitro was not affected by co-culture with normal haemopoietic cells. However, intracellular IL-3 was detected in leukaemic nodules isolated from the liver. Inhibitors specific for IL-3 were not found, although liver-cell conditioned medium and leukaemic nodule sonicates contained potent non-specific inhibitors of cell growth. At 21 days, intracellular IL-3 was also present in spleens and correlated with the presence of non-specific inhibitors. In GVHD, no evidence for IL-3 elaboration in vivo was found, nor did lymphoid populations affected by GVHD spontaneously elaborate it in vitro; however, their competence to produce it was unaffected, as IL-3 was elaborated on subsequent mitogen stimulation in vitro. We also investigated the recovery and circulation of in vitro 111Indium-labelled IL-3 dependent cells after injection in vivo and the half-life of semi-purified IL-3. Dependent cells were not recovered after injection into irradiated recipients, although the cells recirculated for at least 24 hours. Inability to recover dependent cells was explicable on general cytotoxicity which masked potential recovery. The serum half-life of injected partially purified material with IL-3 activity was short (less than 30 min). We conclude that the elaboration of IL-3 by leukaemic WEHI-3b is not an in vitro artifact and these results are discussed in relationship to other growth factors and the leukaemic state, and the origin of IL-3 dependent lines.

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Selected References

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  1. Bazill G. W., Haynes M., Garland J., Dexter T. M. Characterization and partial purification of a haemopoietic cell growth factor in WEHI-3 cell conditioned medium. Biochem J. 1983 Mar 15;210(3):747–759. doi: 10.1042/bj2100747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bindon C., Czerniecki M., Ruell P., Edwards A., McCarthy W. H., Harris R., Hersey P. Clearance rates and systemic effects of intravenously administered interleukin 2 (IL-2) containing preparations in human subjects. Br J Cancer. 1983 Jan;47(1):123–133. doi: 10.1038/bjc.1983.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burgess A. W., Metcalf D. The nature and action of granulocyte-macrophage colony stimulating factors. Blood. 1980 Dec;56(6):947–958. [PubMed] [Google Scholar]
  4. Das S. K., Stanley E. R., Guilbert L. J., Forman L. W. Human colony-stimulating factor (CSF-1) radioimmunoassay: resolution of three subclasses of human colony-stimulating factors. Blood. 1981 Sep;58(3):630–641. [PubMed] [Google Scholar]
  5. 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]
  6. Dexter T. M., Allen T. D., Scott D., Teich N. M. Isolation and characterisation of a bipotential haematopoietic cell line. Nature. 1979 Feb 8;277(5696):471–474. doi: 10.1038/277471a0. [DOI] [PubMed] [Google Scholar]
  7. Dexter T. M., Garland J., Scott D., Scolnick E., Metcalf D. Growth of factor-dependent hemopoietic precursor cell lines. J Exp Med. 1980 Oct 1;152(4):1036–1047. doi: 10.1084/jem.152.4.1036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. English L. S. Immunoregulatory factors produced by activated lymph nodes in vivo. Adv Exp Med Biol. 1982;149:601–607. doi: 10.1007/978-1-4684-9066-4_83. [DOI] [PubMed] [Google Scholar]
  9. Ford W. L., Rolstad B., Fossum S., Hunt S. V., Smith M. E., Sparshott S. M. The stimulus to host cell proliferation in graft-versus-host reactions. Scand J Immunol. 1981 Dec;14(6):705–713. doi: 10.1111/j.1365-3083.1981.tb00613.x. [DOI] [PubMed] [Google Scholar]
  10. Garland J. M., Dexter T. M. 20 alpha-hydroxysteroid dehydrogenase expression in hemopoietic cell cultures and its relationship to interleukin 3. Eur J Immunol. 1982 Dec;12(12):998–1001. doi: 10.1002/eji.1830121203. [DOI] [PubMed] [Google Scholar]
  11. Garland J. M., Dexter T. M. Lymphoid antigens on non-lymphoid factor-dependent haemopoietic cell lines. Adv Exp Med Biol. 1982;149:127–135. doi: 10.1007/978-1-4684-9066-4_18. [DOI] [PubMed] [Google Scholar]
  12. Garland J. M., Dexter T. M. Relationship of haemopoietic growth factor to lymphocytes and interleukin 3: a short review. Lymphokine Res. 1983;2(1):13–22. [PubMed] [Google Scholar]
  13. Gorczynski R. M., Benzing K., MacRae S., Price G. In vitro generation from murine brow marrow cells of accessory cells with discrete antigen presentation capacity and ability to release lymphostimulatory molecules. Immunopharmacology. 1980 Dec;2(4):327–339. doi: 10.1016/0162-3109(80)90017-x. [DOI] [PubMed] [Google Scholar]
  14. Greenberger J. S., Eckner R. J., Ostertag W., Colletta G., Boschetti S., Nagasawa H., Karpas A., Weichselbaum R. R., Moloney W. C. Release of spleen focus-forming virus (SFFV) from differentiation inducible promyelocytic leukemia cell lines transformed in vitro by Friend leukemia virus. Virology. 1980 Sep;105(2):425–435. doi: 10.1016/0042-6822(80)90043-4. [DOI] [PubMed] [Google Scholar]
  15. Greenberger J. S. Self-renewal of factor-dependent hemopoietic progenitor cell-lines derived from long-term bone marrow cultures demonstrates significant mouse strain genotypic variation. J Supramol Struct. 1980;13(4):501–511. doi: 10.1002/jss.400130409. [DOI] [PubMed] [Google Scholar]
  16. Hapel A. J., Lee J. C., Farrar W. L., Ihle J. N. Establishment of continuous cultures of thy1.2+, Lyt1+, 2-T cells with purified interleukin 3. Cell. 1981 Jul;25(1):179–186. doi: 10.1016/0092-8674(81)90242-7. [DOI] [PubMed] [Google Scholar]
  17. Ihle J. N., Keller J., Henderson L., Klein F., Palaszynski E. Procedures for the purification of interleukin 3 to homogeneity. J Immunol. 1982 Dec;129(6):2431–2436. [PubMed] [Google Scholar]
  18. Jones-Villeneuve E. V., Rusthoven J. J., Miller R. G., Phillips R. A. Differentiation of Thy 1-bearing cells from progenitors in long-term bone marrow cultures. J Immunol. 1980 Feb;124(2):597–601. [PubMed] [Google Scholar]
  19. Larsson E. L., Iscove N. N., Coutinho A. Two distinct factors are required for induction of T-cell growth. Nature. 1980 Feb 14;283(5748):664–666. doi: 10.1038/283664a0. [DOI] [PubMed] [Google Scholar]
  20. Lotem J., Sachs L. In vivo inhibition of the development of myeloid leukemia by injection of macrophage- and granulocyte-inducing protein. Int J Cancer. 1981 Sep 15;28(3):375–386. doi: 10.1002/ijc.2910280318. [DOI] [PubMed] [Google Scholar]
  21. Maizel A. L., Mehta S. R., Hauft S., Franzini D., Lachman L. B., Ford R. J. Human T lymphocyte/monocyte interaction in response to lectin: kinetics of entry into the S-phase. J Immunol. 1981 Sep;127(3):1058–1064. [PubMed] [Google Scholar]
  22. Moore M. A., Spitzer G., Metcalf D., Penington D. G. Monocyte production of colony stimulating factor in familial cyclic neutropenia. Br J Haematol. 1974 May;27(1):47–55. doi: 10.1111/j.1365-2141.1974.tb06773.x. [DOI] [PubMed] [Google Scholar]
  23. Raulet D. H., Bevan M. J. A differentiation factor required for the expression of cytotoxic T-cell function. Nature. 1982 Apr 22;296(5859):754–757. doi: 10.1038/296754a0. [DOI] [PubMed] [Google Scholar]
  24. Schrader J. W., Goldschneider I., Bollum F. J., Schrader S. In vitro studies on lymphocyte differentiation. II. Generation of terminal deoxynucleotidyl transferase-positive cells in long-term cultures of mouse bone marrow. J Immunol. 1979 Jun;122(6):2337–2339. [PubMed] [Google Scholar]
  25. Schrader J. W., Lewis S. J., Clark-Lewis I., Culvenor J. G. The persisting (P) cell: histamine content, regulation by a T cell-derived factor, origin from a bone marrow precursor, and relationship to mast cells. Proc Natl Acad Sci U S A. 1981 Jan;78(1):323–327. doi: 10.1073/pnas.78.1.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sekaly R. P., MacDonald H. R., Zaech P., Nabholz M. Cell cycle regulation of cloned cytolytic T cells by T cell growth factor: analysis by flow microfluorometry. J Immunol. 1982 Oct;129(4):1407–1414. [PubMed] [Google Scholar]
  27. Shadduck R. K., Pigoli G., Waheed A., Boegel F. The role of colony-stimulating factor in granulopoiesis. J Supramol Struct. 1980;14(4):423–439. doi: 10.1002/jss.400140403. [DOI] [PubMed] [Google Scholar]
  28. Thakur M. L., Segal A. W., Louis L., Welch M. J., Hopkins J., Peters T. J. Indium-111-labeled cellular blood components: mechanism of labeling and intracellular location in human neutrophils. J Nucl Med. 1977 Oct;18(10):1022–1026. [PubMed] [Google Scholar]
  29. Wagner H., Hardt C., Heeg K., Röllinghoff M., Pfizenmaier K. T-cell-derived helper factor allows in vivo induction of cytotoxic T cells in nu/nu mice. Nature. 1980 Mar 20;284(5753):278–278. doi: 10.1038/284278a0. [DOI] [PubMed] [Google Scholar]
  30. Warner J. F., Dennert G. Effects of a cloned cell line with NK activity on bone marrow transplants, tumour development and metastasis in vivo. Nature. 1982 Nov 4;300(5887):31–34. doi: 10.1038/300031a0. [DOI] [PubMed] [Google Scholar]
  31. Warner N. L., Moore M. A., Metcalf D. A transplantable myelomonocytic leukemia in BALB-c mice: cytology, karyotype, and muramidase content. J Natl Cancer Inst. 1969 Oct;43(4):963–982. [PubMed] [Google Scholar]
  32. Yung Y. P., Moore M. A. Long-term in vitro culture of murine mast cells. III. Discrimination of mast cells growth factor and granulocyte-CSF. J Immunol. 1982 Sep;129(3):1256–1261. [PubMed] [Google Scholar]

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