A murine stromal cell line promotes the proliferation of the human factor-dependent leukemic cell line UT-7.
In long-term human bone marrow cultures, stromal cells of human origin are usually used on the assumption that human primitive progenitor cells do not respond to cytokines produced by stromal cells from other species. There is accumulating evidence, however, that murine stromal cells also promote maintenance and differentiation of very primitive human stem cells, which suggests the existence of novel stromal activities that cross species barriers. In this study, we show that a murine bone marrow-derived stromal cell line, MS-5, allows the proliferation of the human leukemic cell line UT-7. The long-term growth of UT-7 is usually supported only by human interleukin-3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF), or erythropoietin (Epo). None of these three cytokines was involved in the observed effect, since murine GM-CSF and IL-3 do not act on human cells and MS-5 cells do not produce Epo. Soluble stem cell factor (SCF) induced UT-7 cell proliferation. However, S1/S1 mutant fibroblasts also supported UT-7 cell growth and anti-c-kit antibodies only partially abolished UT-7 cell proliferative response to MS-5 cells. These observations excluded a major role of SCF in this system. MS-5-derived growth-promoting activity was diffusible, but attempts to grow UT-7 cells in high levels of known soluble murine stromal-derived cytokines active on human cells showed no or minimal response, suggesting that MS-5's proliferative effect was not mediated by known cytokines. Finally, involvement of an autocrine loop of activation induced by MS-5 was excluded: RT-PCR analysis did not detect increased transcripts for GM-CSF, IL-3, IL-6, SCF, or Epo in UT-7 cells cocultured for 2 to 6 days with MS-5. In addition, UT-7 cell proliferation on MS-5 was not inhibited by neutralizing antibodies against the human GM-CSF receptor or the human IL-6 receptor alpha chain. Whether UT-7 cell proliferation triggered by MS-5 reflects the existence of novel stromal cytokines or results from synergistic interactions on the MS-5 cell surface between extracellular matrix proteins and cytokines will require further investigation.
Auffray
I
I
INSERM U 362, Institut Gustave Roussy, Villejuif, France.
Dubart
A
A
Izac
B
B
Vainchenker
W
W
Coulombel
L
L
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D001483:Base Sequence; D001853:Bone Marrow / Q000737:chemistry / Q000502:physiology; D001854:Bone Marrow Cells; D002450:Cell Communication / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D004247:DNA / Q000032:analysis / Q000235:genetics; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics; D016298:Hematopoietic Cell Growth Factors / Q000235:genetics / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000502:physiology; D006801:Humans; D007377:Interleukin-3 / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics; D007938:Leukemia / Q000473:pathology* / Q000503:physiopathology; D051379:Mice; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / Q000276:immunology; D018123:Receptors, Interleukin / Q000276:immunology; D019947:Receptors, Interleukin-6; D013045:Species Specificity; D019089:Stem Cell Factor; D013997:Time Factors; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D016298:Hematopoietic Cell Growth Factors; D007377:Interleukin-3; D015850:Interleukin-6; D009841:Oligonucleotides; D012333:RNA, Messenger; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor; D018123:Receptors, Interleukin; D019947:Receptors, Interleukin-6; D019089:Stem Cell Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D004247:DNA