Human granulocyte colony-stimulating factor: biologic activities and receptor characterization on hematopoietic cells and small cell lung cancer cell lines

Abstract

Human granulocyte colony-stimulating factor (G-CSF) is a regulatory glycoprotein that stimulates the production of neutrophilic granulocytes from committed hematopoietic progenitor cells both in vitro and in vivo. In this report, we show that biosynthetic (recombinant) human G-CSF enhances colony formation by normal human bone marrow and the human myeloid leukemic cell lines, HL-60 and KG-1, as well as nonhematopoietic small cell lung cancer lines, H128 and H69. G-CSF also modulates multiple differentiated functions of human neutrophils, including enhanced oxidative metabolism in response to f- Met-Leu-Phe (f-MLP), increased antibody-dependent cell-mediated cytotoxicity (ADCC), and augmented arachidonic acid release in response to ionophore and chemotactic agents. These effects are all maximal at a concentration of 100 to 500 pmol/L. Using 125I-labeled recombinant human G-CSF, high affinity binding sites were identified on human neutrophils, the myeloid leukemia cell lines KG-1 and HL-60, and the small cell carcinoma cell lines, H128 and H69. G-CSF receptor numbers ranged between 138 and 285 sites per cell with a kd of 77 to 140 pmol/L, consistent with the concentrations of G-CSF that elicit biologic responses in vitro. Decreased specific binding of 125l-G-CSF by human neutrophils was consistently observed in the presence of excess unlabeled human granulocyte-macrophage colony-stimulating factor (GM-CSF), suggesting competition or down modulation by GM-CSF of the G- CSF receptor.

Population Modeling of Filgrastim PK‐PD in Healthy Adults Following Intravenous and Subcutaneous Administrations

“…The estimated K D value is close to a value of 90.8 pM obtained by us from the in vitro Biacore-based binding assay (data not shown), and in the range of 70 to 150 pM reported elsewhere. 23,29 The estimated ξ value is larger than the number of G-CSF receptors per neutrophil, 50 to 500 reported in the literature. 1 This large value could account for nonspecific binding or underestimation of neutrophils in the bone marrow.…”

Recombinant human granulocyte colony‐stimulating factor inhibits the metastasis of hematogenous and non‐hematogenous tumors in mice

“…Vellenga et al (1987) reported that rhG-CSF stimulates the proliferation of human myeloid leukemia cells in vitro. Avalos et al (1990) showed that rhG-CSF stimulates the in vitro proliferation of non-hematopoietic small-cell lung-cancer cells that possess receptors for G-CSF on the cell surface. , however, reported that although rhG-CSF stimulates in witro proliferation of mouse leukemic cells, it inhibits the development of leukemic cells in vivo.…”

LRG-accelerated differentiation defines unique G-CSFR signaling pathways downstream of PU.1 and C/EBPε that modulate neutrophil activation

“…A specific role for LRG in granulopoiesis has not been elucidated. G-CSF has been shown to be the major regulator of granulopoiesis [4] and supports not only the survival, proliferation, and neutrophilic differentiation of myeloid progenitor cells but also activates certain functions of mature, terminally differentiated neutrophils [5][6][7]. The biological activities of G-CSF are mediated by specific receptors on the surface of responsive cells.…”

“…Studies in mice with knockout or knock-in mutations in the G-CSFR gene suggest that the G-CSFR generates unique, nonredundant signals required for polymorphonuclear neutrophil (PMN) production and marrow egress to maintain homeostatic levels of circulating PMN [7][8][9][10]. G-CSFR knockout mice have chronic neutropenia with a uniform decrease in myeloid cells in the bone marrow.…”