IL‐3 specifically inhibits GM‐CSF binding to the higher affinity receptor

Taketazu, Fumitoshi; Shibuya, Kengo; Kuwaki, Tomoaki; Tsumura, Haruhiko; Miyazono, Kohei; Miyagawa, Kiyoshi; Takaku, Fumimaro

doi:10.1002/jcp.1041460209

Cited by 32 publications

(9 citation statements)

References 19 publications

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“…8-fold increase in WCC during the second week of treatment and a 1 . In fact, an inhibition of the effect of GM-CSF by activation of the IL-3 receptor has been reported in cell lines (Taketazu et al, 1991). The platelet count continued to rise after discontinuation of treatment as was the case with most of our patients.…”

Section: Discussionsupporting

confidence: 75%

The effect of the GM‐CSF/IL‐3 fusion protein PIXY321 on bone marrow and circulating haemopoietic cells of previously untreated patients with cancer

et al. 1996

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This is a phase I/II study of the GM-CSF/IL-3 fusion protein (PIXY321. Patients were treated with PIXY321 at a daily subcutaneous dose of 500, 750 and 1000 micrograms/m2 for 14 d. Side-effects were mild and consisted mainly of injection-site reactions and constitutional symptoms. A biphasic modest increase of white blood count (2-5-fold) and platelets (1-1.5 fold) was seen, accompanied by an increased bone marrow cellularity and an increase in circulating progenitors. Colony-forming cells in the blood rose to a median of 184 granulocyte/macrophage-colony forming cells (GM-CFC)/ml, eight Mix-CFC/ml, 250 burst forming units-erythroid (BFU-E)/ml and 140 CFU-mega-karyocytes/ml, corresponding to a 10-, 2.5, 8- and 30-fold increase respectively. When seeded for long-term culture on irradiated bone marrow stroma, the mobilized cells were not able to sustain haemopoiesis in vitro to the same degree as bone marrow. Taken together these results indicate that PIXY321 has a biological effect in humans more similar to that of IL-3 than to that of GM-CSF.

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Section: Discussionsupporting

confidence: 75%

The effect of the GM‐CSF/IL‐3 fusion protein PIXY321 on bone marrow and circulating haemopoietic cells of previously untreated patients with cancer

et al. 1996

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“…The importance of the (3 subunit is emphasized by the fact that it is shared by three different receptors with clear specificities for their respective ligands, GM-CSF, interleukin 3, and interleukin 5. These receptors possess overlapping functional characteristics, and more than one receptor may be expressed simultaneously in a given cell type (19,(40)(41)(42)(43)(44)(45)(46)(47)(48). The a subunit of each receptor may play a role in defining the specificity of the cellular response triggered by the binding of the respective ligand.…”

Section: Discussionmentioning

confidence: 99%

The alpha subunit of the human granulocyte-macrophage colony-stimulating factor receptor signals for glucose transport via a phosphorylation-independent pathway.

Ding

Rivas²,

Heaney³

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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The receptor for granulocyte-macrophage colony-stimulating factor (GM-CSF) is composed of an a and B3 subunit, which together form the high-affinity receptor. The a subunit by itself binds ligand at low affinity, whereas the isolated (8 subunit does not bind GM-CSF. It is generally believed that the high-affinity receptor is responsible for the multiple functions of GM-CSF and that the isolated a subunit (GMRa) does not transduce a signal. Xenopus laevis oocytes injected with RNA encoding human GMRa expressed up to 1010 low-affinity sites for GM-CSF (Kd = 6 nM). GM-CSF binding to the a subunit expressed in Xenopus oocytes caused activation of 2-deoxyglucose transport through endogenous glucose transporters. 2-Deoxyglucose transport was stimulated by similar low concentrations of GM-CSF in HL-60 leukemia cells as well as normal human neutrophils and Xenopus oocytes expressing GMRa. Engagement of the isolated a subunit in oocytes did not lead to protein phosphorylation or tyrosine phosphorylation of mitogen-activated protein kinase (MAP kinase). Staurosporin and genistein inhibited GM-CSFinduced tyrosine phosphorylation of MAP kinase in human neutrophils and HL-60 cells without affecting GM-CSFstimulated uptake of 2-deoxyglucose. These results provide direct evidence that the isolated a subunit signals for hexose transport and can do so without engagement of the kinase cascade. Our data also indicate that signaling for hexose uptake may occur in a phosphorylation-independent manner in cells expressing the high-affinity GM-CSF receptor.The receptor for granulocyte-macrophage colony-stimulating factor (GM-CSF) is composed of an a and ( subunit, and the a subunit (GMRa) by itselfbinds ligand at low affinity (Kd = 3-7 nM) (1-6). The isolated 8 subunit (GMR(3) does not bind GM-CSF; however, it associates with GMRa to form a high-affinity receptor (Kd = 20-40 pM) (3-6). GM-CSF receptors are present on myeloid progenitors and mature neutrophils, eosinophils, and mononuclear phagocytes (7)(8)(9)(10)(11)(12)(13). GMRa is also present in some nonhematopoietic cells, such as placenta and melanoma cells (1,14,15), while the GMR(8 is mainly restricted to hematopoietic cells (3-5, 12).It is generally believed that the biological functions exerted by GM-CSF are mediated by the high-affinity receptor and that the isolated GMRa does not transduce a signal (3-6, 16, 17). GMRa, however, binds GM-CSF and in association with GMR,8 transduces signals involving protein phosphorylation (2,(18)(19)(20). To examine the functional capability of the isolated a subunit, we expressed it in Xenopus laevis oocytes and discovered that it could signal for glucose transport without involving the phosphorylation of mitogen-activated protein kinase (MAP kinase). MATERIALS AND METHODSComplementary DNAs for the human GMRa (21) and the human glucose transporter 4 (GLUT4) (22) cloned in pBluescript (Stratagene) were linearized by digestion with restriction enzymes, and the sense or antisense RNA was transcribed in vitro (23). Stage V and VI oocy...

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“…We have also observed this phenomenon using the ceIls with both GM-CSF receptors and 1L-3 receptors [50]. Furthermore, we have discovered that, on both KG-1 cells and TF-1 cells, this cross-reactivity with GM-CSF and IL-3 specifically occurred on the P-chain of GM-CSF receptors in cross-linking studies 1501.…”

Section: Discussionmentioning

confidence: 69%

Structural and functional analyses of glycosylation on the distinct molecules of human GM‐CSF receptors

Shibuya

Miyagawa

Kitamura

et al. 1991

European Journal of Biochemistry

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We have previously demonstrated that granulocyte-macrophage colony-stimulating factor (GM-CSF) receptors are composed of at least two molecules of 80 and 135 kDa, which were denoted a-and /?-chains, respectively [Chiba, S., Shibuya, K., Piao, Y.-F., Tojo, A., Sasaki, N., Matsuki, S., Miyagawa, K., Cell Regul. 1,. In this paper, we describe an investigation of the biochemical disparity noted between the a-and /?-chains of GM-CSF receptors using proteolytic and deglycosidic enzymes, and further demonstrate the potential importance of carbohydrate structures of the GM-CSF receptors using different lectins and glycoprotein synthesis inhibitors. Cross-linked a-and /?-chains with 251-GM-CSF were digested by Staphylococcus aureus V8 protease and gave a different pattern. Furthermore, the size of the a-chain was reduced by 25 kDa by the removal of the N-linked oligosaccharides with peptidase: N-glycosidase F treatment, whereas that of the /?-chain remained unmodified by the enzyme. These results suggest that the a-chain of GM-CSF receptors agrees with the recently cloned low-affinity GM-CSF receptor [Gearing, D. P., King, J. A,, Gough, N. M. & Nicola, N. A. (1989) EMBO J. 8, 3667-36761 having approximately 30% N-linked oligosaccharides and is biochemically different from the a/?-chain. By analyses using lectins, some of the oligosaccharides in the CIchain seem to be the complex-type and/or hybrid-type, because wheat germ agglutinin and leukoagglutinating phytohemagglutinin inhibited both GM-CSF-induced proliferation and GM-CSF binding to its receptors. Further analyses using glycoprotein synthesis inhibitors showed that N-linked processing of the a-chain, especially glucose removal by glucosidase I and I1 (whose activities are inhibited by deoxynojirimycin), appeared to be required for the expression onto the cell surface although the /3-chain expression was little affected by their inhibitors. Thus the /?-chain, probably located near the a-chain on the cell surface, was associated with a high-affinity class of GM-CSF receptors.Granulocyte-macrophage colony-stimulating factor (GM-CSF) is one of the colony-stimulating factors (CSFs) for hematopoietic progenitor cells. Most of the CSFs were purified, identified and their cDNAs were cloned. Biochemical and biological characteristics of GM-CSF receptors have been reported. Previously, only a single affinity class of GM-CSF receptor was shown on human hematopoietic cells [9 -121. However, our recent extensive studies showed that most hematopoietic cells expressed both a highaffinity class of binding site with a Kd of 10-50 pM and a low-affinity one with a Kd of 0.9 -2 nM [13 ~ 151. Chemical cross-linking analyses further revealed that the GM-CSF receptors consisted of at least two molecules of 80 kDa and 135 kDa [14, 151. We also showed that nonhematopoietic cells had only a low-affinity receptor, with a Kd of 1.5-10 nM, whose molecular mass was approximately 65 -85 kDa, and could not have their growth modified by GM-CSF [16].In our previous report, the 80-kDa and 135-kDa ...

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IL‐3 specifically inhibits GM‐CSF binding to the higher affinity receptor

Cited by 32 publications

References 19 publications

The effect of the GM‐CSF/IL‐3 fusion protein PIXY321 on bone marrow and circulating haemopoietic cells of previously untreated patients with cancer

The effect of the GM‐CSF/IL‐3 fusion protein PIXY321 on bone marrow and circulating haemopoietic cells of previously untreated patients with cancer

The alpha subunit of the human granulocyte-macrophage colony-stimulating factor receptor signals for glucose transport via a phosphorylation-independent pathway.

Structural and functional analyses of glycosylation on the distinct molecules of human GM‐CSF receptors

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