Molecular cloning and expression of cDNA for human granulocyte colony-stimulating factor

Abstract

Granulocyte colony-stimulating factor (G-CSF) is a member of the CSF family of hormone-like glycoproteins that regulate haematopoietic cell proliferation and differentiation, and G-CSF almost exclusively stimulates the colony formation of granulocytes from committed precursor cells in semi-solid agar culture. Recently, Nomura et al. have established a human squamous carcinoma cell line (designated CHU-2) from a human oral cavity tumour which produces large quantities of CSF constitutively, and the CSF produced by CHU-2 cells has been purified to homogeneity from the conditioned medium. We have now determined the partial amino-acid sequence of the purified G-CSF protein, and by using oligonucleotides as probes, have isolated several clones containing G-CSF complementary DNA from the cDNA library prepared with messenger RNA from CHU-2 cells. The complete nucleotide sequences of two of these cDNAs were determined and the expression of the cDNA in monkey COS cells gave rise to a protein showing authentic G-CSF activity. Furthermore, Southern hybridization analysis of DNA from normal leukocytes and CHU-2 cells suggests that the human genome contains only one gene for G-CSF and that some rearrangement has occurred within one of the alleles of the G-CSF gene in CHU-2 cells.

Prior chemotherapy does not prevent effective mobilisation by G-CSF of peripheral blood progenitor cells

“…The characterisation (Nicola et al, 1979) and molecular cloning of this molecule (Nagata et al, 1986) has allowed its evaluation in a variety of clinical settings including for example, administration of G-CSF to patients following high dose chemotherapy and bone marrow transplantation (Sheridan et al, 1989).…”

Anti‐G presenting as a blood transfusion error

“…The question, whether these mutations are solely responsible for the neutropenia cannot be answered. Until now, all relevant splice sites in the G-CSF gene were found in the canonical GT-AG donor-acceptor sites [22]. Since alternative splicing is a known phenomenon in biologically active G-CSF molecules [22,23], at least the variants in intron 4 could be a reason for a modified G-CSF profile in this patient.…”

“…Until now, all relevant splice sites in the G-CSF gene were found in the canonical GT-AG donor-acceptor sites [22]. Since alternative splicing is a known phenomenon in biologically active G-CSF molecules [22,23], at least the variants in intron 4 could be a reason for a modified G-CSF profile in this patient. Splicing variants with a lowered affinity to the G-CSF-receptor were previously described [23,24].…”

“…Homozygous base exchanges were found in the patient's G-CSF gene when compared to the originally described sequence [21,22].…”

Allogeneic Bone Marrow Transplantation for Malignant Hematologic Disorders in Children.

“…cloned and its clinical use became possible in Japan as well as United States and European countries (Nagata et al 1986; Souza et al 1986). rG-CSF is expected to induce a rapid recovery of granulopoiesis from severe granulocytopenia and eventually to decrease infectious complications.…”