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Constitutive expression of granulocyte‐colony stimulating factor receptor on a human B‐lymphoblastoid cell line
Author(s) -
Morikawa Keiko,
Morikawa Shigeru,
Miyawaki Toshio,
Nagasaki Makoto,
Torii Ikuko,
Imai Katsuyuki
Publication year - 1996
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.1966.d01-1806.x
Subject(s) - microbiology and biotechnology , cell culture , receptor , biology , cell surface receptor , biotinylation , trypan blue , immunofluorescence , receptor expression , flow cytometry , cell , antibody , biochemistry , immunology , genetics
The present study demonstrated that a human B‐ cell line derived from non‐Hodgkin's lymphoma, HCF‐MLpN, constitutively expressed G‐CSF receptor on the cell surface. G‐CSF binding to the cell surface was shown by immunofluorescence staining using biotinylated G‐CSF preparation and analysed by flow cytometry. Specific binding of G‐CSF to the cells was shown by pretreatment with unlabelled G‐CSF. In the radioreceptor assay and Scatchard plot analysis using radiolabelled ligand, MLpN cells revealed a single species of binding site with an equilibrium dissociation constant of 167 (153–182) p M and a maximal binding site per cell of 1076 (1044–1116). The G‐CSF receptor mRNA transcript was exhibited in the RNA from MLpN cells by reverse transcriptase polymerase chain reaction procedure. [ 3 H]thymidine incorporation and trypan blue exclusion showed that the G‐CSF receptor was capable of transducing the growth signal to HCF‐MLpN cells. A small fraction of fresh B blasts from six patients with B‐cell lymphoma and leukaemia displayed G‐CSF binding by two‐colour immunofluorescence staining. In contrast, a panel of seven B‐cell lines was negative for the binding to biotinylated G‐CSF preparation. These results suggest that the phenotype of G‐CSF binding may be lost during the culture. The expression of G‐CSF receptor in HCF‐MLpN cells appeared to be exceptional.