scholarly journals Human Fibroblasts(KMST-6/RAS Cell Line) Transformed with 60Co Gamma-rays and c-Ha-ras Oncogene Produce a Large Amount of Granulocyte Colony-stimulating Factor(G-CSF); Production is Enhanced by cAMP, Theophylline, and Butyrate.

1995 ◽  
Vol 20 (1) ◽  
pp. 41-45 ◽  
Author(s):  
Kunihiro Kawashima ◽  
Israt Jahan ◽  
Kazuo Fushimi ◽  
Koichiro Mihara ◽  
Masayoshi Namba
Keyword(s):  
Cell Line ◽  
Gamma Rays ◽  
Human Fibroblasts ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Ras Oncogene ◽  
60Co Gamma ◽  
60Co Gamma Rays ◽  
Stimulating Factor ◽  
Csf Production

scholarly journals Adhesion of NFS-60 myeloid leukemia cells to MC3T3-G2/PA6 stromal cells induces granulocyte colony-stimulating factor production

Blood ◽  
1994 ◽  
Vol 84 (2) ◽  
pp. 415-420 ◽  
Author(s):  
T Yoshikubo ◽  
K Ozawa ◽  
K Takahashi ◽  
M Nishikawa ◽  
N Horiuchi ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Cell Line ◽  
Cell Lines ◽  
Stromal Cells ◽  
Myeloid Leukemia ◽  
Myeloid Cell ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Stimulating Factor ◽  
Csf Production

Abstract To examine the interaction between immature myeloid cells and stromal cells in the induction of granulocyte colony-stimulating factor (G-CSF) production, stromal cells of the MC3T3-G2/PA6 (PA6) murine cell line, which has preadipocyte characteristics and can support hematopoiesis, were cocultured with various myeloid cell lines and G-CSF mRNA expression was examined by Northern and reverse transcriptase- polymerase chain reaction analyses. A significant amount of G-CSF mRNA was induced by the culture of an interleukin-3/G-CSF-dependent murine myeloid leukemia cell line, NFS-60, on PA6 stromal cells for 16 hours. Using a G-CSF-dependent subline of DA-1 (DA-1N), the biologic activity of G-CSF was also detected in PA6/NFS-60 coculture supernatants, but not in the culture supernatant of PA6 or NFS-60 alone. Direct contact of NFS-60 cells with the PA6 stromal layer was essential for the induction of G-CSF mRNA, as indicated by the following observations: (1) NFS-60 cells efficiently adhered to PA6 cells; (2) medium conditioned by NFS-60 cells did not contain the activity to induce G- CSF mRNA in PA6 cells; and (3) induction of G-CSF mRNA was not observed when NFS-60 cells were separated from PA6 cells by a microporous membrane (0.45-microns pore size). Several other myeloid cell lines, including FDC-P2, 32Dcl3, WEHI-3, and DA-1, did not induce G-CSF mRNA expression after the coculture with PA6 cells, although significant numbers of these cells adhered to PA6 cells. Therefore, NFS-60 cells may express or overexpress a molecule that is involved in adhesion- mediated induction of G-CSF production.

scholarly journals Adhesion of NFS-60 myeloid leukemia cells to MC3T3-G2/PA6 stromal cells induces granulocyte colony-stimulating factor production

Blood ◽  
1994 ◽  
Vol 84 (2) ◽  
pp. 415-420 ◽  
Author(s):  
T Yoshikubo ◽  
K Ozawa ◽  
K Takahashi ◽  
M Nishikawa ◽  
N Horiuchi ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Cell Line ◽  
Cell Lines ◽  
Stromal Cells ◽  
Myeloid Leukemia ◽  
Myeloid Cell ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Stimulating Factor ◽  
Csf Production

To examine the interaction between immature myeloid cells and stromal cells in the induction of granulocyte colony-stimulating factor (G-CSF) production, stromal cells of the MC3T3-G2/PA6 (PA6) murine cell line, which has preadipocyte characteristics and can support hematopoiesis, were cocultured with various myeloid cell lines and G-CSF mRNA expression was examined by Northern and reverse transcriptase- polymerase chain reaction analyses. A significant amount of G-CSF mRNA was induced by the culture of an interleukin-3/G-CSF-dependent murine myeloid leukemia cell line, NFS-60, on PA6 stromal cells for 16 hours. Using a G-CSF-dependent subline of DA-1 (DA-1N), the biologic activity of G-CSF was also detected in PA6/NFS-60 coculture supernatants, but not in the culture supernatant of PA6 or NFS-60 alone. Direct contact of NFS-60 cells with the PA6 stromal layer was essential for the induction of G-CSF mRNA, as indicated by the following observations: (1) NFS-60 cells efficiently adhered to PA6 cells; (2) medium conditioned by NFS-60 cells did not contain the activity to induce G- CSF mRNA in PA6 cells; and (3) induction of G-CSF mRNA was not observed when NFS-60 cells were separated from PA6 cells by a microporous membrane (0.45-microns pore size). Several other myeloid cell lines, including FDC-P2, 32Dcl3, WEHI-3, and DA-1, did not induce G-CSF mRNA expression after the coculture with PA6 cells, although significant numbers of these cells adhered to PA6 cells. Therefore, NFS-60 cells may express or overexpress a molecule that is involved in adhesion- mediated induction of G-CSF production.

scholarly journals Antiproliferative and differentiative effects of recombinant interleukin-4 on a granulocyte colony-stimulating factor-dependent myeloblastic leukemic cell line

Blood ◽  
1991 ◽  
Vol 78 (2) ◽  
pp. 471-478
Author(s):  
Y Imai ◽  
N Nara ◽  
S Tohda ◽  
K Nagata ◽  
T Suzuki ◽  
...  
Keyword(s):  
Cell Line ◽  
Leukemic Cell ◽  
Interleukin 4 ◽  
Tnf Alpha ◽  
Adherent Cells ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Leukemic Cell Line ◽  
Nonadherent Cells ◽  
Stimulating Factor

The effect of recombinant human interleukin-4 (IL-4) on a granulocyte colony-stimulating factor (G-CSF)-dependent human myeloblastic leukemic cell line, OCI-AML1a, was investigated. IL-4 suppressed the clonogenic cell growth in methylcellulose culture, inhibited the uptake of 3H thymidine in a dose-dependent manner at 5 to 100 U/mL, and consequently suppressed the growth of clonogenic cells in short- and long-term suspension cultures. In addition, IL-4 markedly increased the number of adherent cells. These adherent cells were alpha-naphthyl-butyrate (alpha-NB) esterase-positive and showed macrophage-like appearance, increased expression of CD14, CD11b, CD23, and Ia, and significantly decreased clonogenicity. On the other hand, nonadherent cells growing in suspension showed only slight increase in proportion of alpha-NB esterase-positive or monocyte/macrophage-like cells and increased CD23 expression by an addition of IL-4. The clonogenicity of the nonadherent cells was not significantly influenced by IL-4. By addition of the media conditioned by OCI-AML1a cells in the presence of IL-4, the clonogenic cells growth of OCIAML1a cells was suppressed and adherent cells were markedly increased. The suppressive and differentiative effects on OCI/AML1a cells of the conditioned media and IL-4 itself were almost completely abolished by anti-IL-4 antibody. Furthermore, the neutralizing antibodies against transforming growth factor-beta 2 (TGF-beta 2), tumor necrosis factor-alpha (TNF-alpha), or IL-6 did not influence the effect of recombinant IL-4. Taken together, IL-4 was shown to suppress the growth and induce differentiation toward adherent macrophage-like cells of the G-CSF-dependent myeloblastic cell line. The effect of IL-4 may be direct, and not secondary via inducing production of other cytokines such as TGF-beta, TNF-alpha, or IL-6 by leukemic cells.

scholarly journals Antiproliferative and differentiative effects of recombinant interleukin-4 on a granulocyte colony-stimulating factor-dependent myeloblastic leukemic cell line

Blood ◽  
1991 ◽  
Vol 78 (2) ◽  
pp. 471-478 ◽  
Author(s):  
Y Imai ◽  
N Nara ◽  
S Tohda ◽  
K Nagata ◽  
T Suzuki ◽  
...  
Keyword(s):  
Cell Line ◽  
Leukemic Cell ◽  
Interleukin 4 ◽  
Tnf Alpha ◽  
Adherent Cells ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Leukemic Cell Line ◽  
Nonadherent Cells ◽  
Stimulating Factor

Abstract The effect of recombinant human interleukin-4 (IL-4) on a granulocyte colony-stimulating factor (G-CSF)-dependent human myeloblastic leukemic cell line, OCI-AML1a, was investigated. IL-4 suppressed the clonogenic cell growth in methylcellulose culture, inhibited the uptake of 3H thymidine in a dose-dependent manner at 5 to 100 U/mL, and consequently suppressed the growth of clonogenic cells in short- and long-term suspension cultures. In addition, IL-4 markedly increased the number of adherent cells. These adherent cells were alpha-naphthyl-butyrate (alpha-NB) esterase-positive and showed macrophage-like appearance, increased expression of CD14, CD11b, CD23, and Ia, and significantly decreased clonogenicity. On the other hand, nonadherent cells growing in suspension showed only slight increase in proportion of alpha-NB esterase-positive or monocyte/macrophage-like cells and increased CD23 expression by an addition of IL-4. The clonogenicity of the nonadherent cells was not significantly influenced by IL-4. By addition of the media conditioned by OCI-AML1a cells in the presence of IL-4, the clonogenic cells growth of OCIAML1a cells was suppressed and adherent cells were markedly increased. The suppressive and differentiative effects on OCI/AML1a cells of the conditioned media and IL-4 itself were almost completely abolished by anti-IL-4 antibody. Furthermore, the neutralizing antibodies against transforming growth factor-beta 2 (TGF-beta 2), tumor necrosis factor-alpha (TNF-alpha), or IL-6 did not influence the effect of recombinant IL-4. Taken together, IL-4 was shown to suppress the growth and induce differentiation toward adherent macrophage-like cells of the G-CSF-dependent myeloblastic cell line. The effect of IL-4 may be direct, and not secondary via inducing production of other cytokines such as TGF-beta, TNF-alpha, or IL-6 by leukemic cells.

scholarly journals Dexamethasone and 1,25-dihydroxyvitamin D3, but not cyclosporine A, inhibit production of granulocyte-macrophage colony-stimulating factor in human fibroblasts

Blood ◽  
1991 ◽  
Vol 77 (9) ◽  
pp. 1912-1918 ◽  
Author(s):  
A Tobler ◽  
HP Marti ◽  
C Gimmi ◽  
AB Cachelin ◽  
S Saurer ◽  
...  
Keyword(s):  
Human Fibroblasts ◽  
Tnf Alpha ◽  
Colony Stimulating Factor ◽  
Dihydroxyvitamin D3 ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Csf Production

Abstract Tumor necrosis factor alpha (TNF alpha) stimulates granulocyte- macrophage colony-stimulating factor (GM-CSF) production in human fibroblasts and other mesenchymal cells. However, relatively little is known about agents that downregulate cytokine production in these cells. In the present report we show that dexamethasone (Dexa), a synthetic glucocorticoid, markedly reduced GM-CSF production in TNF alpha-stimulated fibroblasts at both the protein and the RNA levels. CSF activity, GM-CSF protein, and RNA levels, determined by an in vitro colony-forming assay in normal human bone marrow cells, by an enzyme immunoassay, and by Northern blotting assay, were reduced to greater than 90% of control values by Dexa (1 mumol/L). Similarly, 1,25- dihydroxyvitamin D3 [1,25(OH)2D3], a hormone with possible physiologic immunoregulatory significance, reduced GM-CSF expression in a concentration- and time-dependent manner. However, this repression was less pronounced than that of Dexa, and in part due to a decreased proliferative activity. In contrast, cyclosporine A (CsA), another immunosuppressive agent, did not alter GM-CSF expression in TNF alpha- stimulated fibroblasts. Our in vitro studies suggest that by inhibiting GM-CSF production in fibroblasts, glucocorticoids and possibly 1,25(OH)2D3, but not CsA, may attenuate TNF alpha-mediated inflammatory processes and influence the regulation of hematopoiesis.

scholarly journals Granzyme B and perforin lytic proteins are expressed in CD34+ peripheral blood progenitor cells mobilized by chemotherapy and granulocyte colony-stimulating factor

Blood ◽  
1995 ◽  
Vol 86 (9) ◽  
pp. 3500-3506 ◽  
Author(s):  
C Berthou ◽  
JP Marolleau ◽  
C Lafaurie ◽  
A Soulie ◽  
L Dal Cortivo ◽  
...  
Keyword(s):  
Cell Line ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Granzyme B ◽  
Cellular Adhesion ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Hematopoietic Stem ◽  
Cd34 Cells ◽  
Stimulating Factor

Granzyme B and perforin are cytoplasmic granule-associated proteins used by cytotoxic T lymphocytes and natural killer (NK) cells to kill their targets. However, granzyme B gene expression has also been detected in a non-cytotoxic hematopoietic murine multipotent stem cell line, FDCP-Mix. The objective of the present study was to investigate whether granzyme B and perforin could be expressed in human hematopoietic CD34+ cells and if present, discover what their physiologic relevance could be. The primitive CD34+ human cell line KG1a was investigated first and was found to express granzyme B and perforin. Highly purified hematopoietic stem/progenitor cells were then selected using the CD34 surface antigen as marker. Steady-state bone marrow (BM) CD34+ cells did not contain these proteins. Peripheral blood (PB) CD34+ cells, which had been induced to circulate, were also analyzed. After chemotherapy (CT) and granulocyte colony-stimulating factor (G-CSF) treatment, CD34+ cells strongly expressed mRNAs and proteins of granzyme B and perforin. In contrast, CD34+ cells mobilized by G-CSF alone were negative. Western blot analysis further showed that granzyme B and perforin proteins were identical in CD34+ cells and activated PBLs. Such proteins might be implicated in the highly efficient migration of CD34+ stem/progenitor cells from BM to PB after CT and G-CSF treatment. The cellular adhesion mechanisms involved in the BM homing of CD34+ cells are disrupted at least temporarily after CT. The Asp-ase proteolytic activity of granzyme B on extracellular matrix proteins could be used by progenitor cells for their rapid detachment from BM stromal cells and perforin might facilitate their migration across the endothelial cell barrier.

Telomere elongation observed in immortalized human fibroblasts by treatment with 60Co gamma rays or 4-nitroquinoline 1-oxide

1996 ◽  
Vol 97 (1) ◽  
Author(s):  
Shinsuke Sugihara ◽  
Koichiro Mihara ◽  
Tohru Marunouchi ◽  
Hajime Inoue ◽  
Masayoshi Namba
Keyword(s):  
Gamma Rays ◽  
Human Fibroblasts ◽  
Telomere Elongation ◽  
60Co Gamma ◽  
60Co Gamma Rays
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