Peripheral Blood Neutrophils of Germ-Free Rats Modified by In Vivo Granulocyte-Colony-Stimulating Factor and Exposure to Natural Environment

Abstract Recombinant rat stem cell factor (rrSCF) and recombinant human granulocyte colony-stimulating factor (G-CSF) coinjected for 1 week in rats cause a synergistic increase in mature marrow neutrophils accompanied by a striking decrease in erythroid and lymphoid marrow elements. The spleens of the same rats show increased granulopoiesis as well as increased erythropoiesis as compared with the spleens of rats treated with either growth factor alone. Splenic extramedullary erythropoiesis may act to compensate for the decrease in marrow erythropoiesis. The coinjection of rrSCF and G-CSF causes an increase in marrow mast cells at the end of 1 week, but the increase is much less than in rrSCF-alone-treated rats. The combination of rrSCF and G- CSF increases the rate of release of marrow neutrophils into the circulation and causes a dramatic synergistic peripheral neutrophilia, beginning especially after 4 days of treatment. Colony-forming assays of all experimental groups showed a synergistic increase in colony- forming unit granulocyte-macrophage (CFU-GM) in the marrow, but not in peripheral blood, after coincubation with SCF plus granulocyte- macrophage CSF (GM-CSF) as opposed to GM-CSF alone, showing anatomic compartmentalization between a more primitive marrow CFU-GM subset and a more mature peripheral blood CFU-GM subset. In vivo daily administration of SCF plus GM-CSF results in a synergistic increase in marrow neutrophils, but not the striking synergistic increase in circulating neutrophils that is observed with SCF plus G-CSF.

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Recombinant rat stem cell factor synergizes with recombinant human granulocyte colony-stimulating factor in vivo in mice to mobilize peripheral blood progenitor cells that have enhanced repopulating potential

Blood ◽

10.1182/blood.v82.6.1720.1720 ◽

1993 ◽

Vol 82 (6) ◽

pp. 1720-1723 ◽

Cited By ~ 91

Author(s):

RA Briddell ◽

CA Hartley ◽

KA Smith ◽

IK McNiece

Keyword(s):

Stem Cell ◽

Progenitor Cells ◽

Peripheral Blood ◽

Stem Cell Factor ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

Peripheral Blood Progenitor Cells ◽

Stimulating Factor ◽

Synergistic Increase

Abstract Splenectomized mice treated for 7 days with pegylated recombinant rat stem cell factor (rrSCF-PEG) showed a dose-dependent increase in peripheral blood progenitor cells (PBPC) that have enhanced in vivo repopulating potential. A dose of rrSCF-PEG at 25 micrograms/kg/d for 7 days produced no significant increase in PBPC. However, when this dose of rrSCF-PEG was combined with an optimal dose of recombinant human granulocyte colony-stimulating factor (rhG-CSF; 200 micrograms/kg/d), a synergistic increase in PBPC was observed. Compared with treatment with rhG-CSF alone, the combination of rrSCF-PEG plus rhG-CSF resulted in a synergistic increase in peripheral white blood cells, in the incidence and absolute numbers of PBPC, and in the incidence and absolute numbers of circulating cells with in vivo repopulating potential. These data suggest that low doses of SCF, which would have minimal, if any, effects in vivo, can synergize with optimal doses of rhG-CSF to enhance the mobilization of PBPC stimulated by rhG-CSF alone.

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Effect of the in vivo application of granulocyte colony‐stimulating factor on NK cells in bone marrow and peripheral blood

Journal of Cellular and Molecular Medicine ◽

10.1111/jcmm.13539 ◽

2018 ◽

Vol 22 (6) ◽

pp. 3025-3034 ◽

Cited By ~ 4

Author(s):

Xing‐Xing Yu ◽

Ting‐Ting Han ◽

Ling‐Ling Xu ◽

Ying‐Jun Chang ◽

Xiao‐Jun Huang ◽

...

Keyword(s):

Bone Marrow ◽

Nk Cells ◽

Peripheral Blood ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

Stimulating Factor

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Recombinant rat stem cell factor synergizes with recombinant human granulocyte colony-stimulating factor in vivo in mice to mobilize peripheral blood progenitor cells that have enhanced repopulating potential

Blood ◽

10.1182/blood.v82.6.1720.bloodjournal8261720 ◽

1993 ◽

Vol 82 (6) ◽

pp. 1720-1723 ◽

Cited By ~ 3

Author(s):

RA Briddell ◽

CA Hartley ◽

KA Smith ◽

IK McNiece

Keyword(s):

Stem Cell ◽

Progenitor Cells ◽

Peripheral Blood ◽

Stem Cell Factor ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

Peripheral Blood Progenitor Cells ◽

Stimulating Factor ◽

Synergistic Increase

Splenectomized mice treated for 7 days with pegylated recombinant rat stem cell factor (rrSCF-PEG) showed a dose-dependent increase in peripheral blood progenitor cells (PBPC) that have enhanced in vivo repopulating potential. A dose of rrSCF-PEG at 25 micrograms/kg/d for 7 days produced no significant increase in PBPC. However, when this dose of rrSCF-PEG was combined with an optimal dose of recombinant human granulocyte colony-stimulating factor (rhG-CSF; 200 micrograms/kg/d), a synergistic increase in PBPC was observed. Compared with treatment with rhG-CSF alone, the combination of rrSCF-PEG plus rhG-CSF resulted in a synergistic increase in peripheral white blood cells, in the incidence and absolute numbers of PBPC, and in the incidence and absolute numbers of circulating cells with in vivo repopulating potential. These data suggest that low doses of SCF, which would have minimal, if any, effects in vivo, can synergize with optimal doses of rhG-CSF to enhance the mobilization of PBPC stimulated by rhG-CSF alone.

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In Vivo Effects of Flt3/Flk2 Ligand on Mobilization of Hematopoietic Progenitors in Primates and Potent Synergistic Enhancement With Granulocyte Colony-Stimulating Factor

Blood ◽

10.1182/blood.v90.2.620.620_620_629 ◽

1997 ◽

Vol 90 (2) ◽

pp. 620-629 ◽

Cited By ~ 1

Author(s):

Thalia Papayannopoulou ◽

Betty Nakamoto ◽

Robert G. Andrews ◽

Stewart D. Lyman ◽

Minako Y. Lee

Keyword(s):

Functional Properties ◽

Peripheral Blood ◽

Combined Treatment ◽

Mast Cell Activation ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

Synergistic Enhancement ◽

Stimulating Factor

The Flt3 receptor is expressed in primitive hematopoietic cells and its ligand exerts proliferative effects on these cells in vitro in synergy with other cytokines. To expand on the functional properties of Flt3 ligand (FL) in vivo we treated nonhuman primates with FL and tested its ability to mobilize stem/progenitor cells when given alone or in combination with granulocyte colony-stimulating factor (G-CSF ) treatment. FL alone (200 μg/kg/day) mobilizes progenitors with slow kinetics and with a peak effect at the end of 2 weeks of treatment. The spectrum of mobilized progenitors includes myeloid, lymphoid, megakaryocytic, and osteoclastogenic but a low proportion of burst-forming unit (BFU)e. Bone marrow (BM) studies before and during the treatment suggested that proliferative effects in BM may have preceded effects on peripheral blood mobilization. To assess the synergy of FL with G-CSF in mobilization of progenitors we used two schemes: one in which G-CSF was used for the last 5 days of a 12-day treatment with FL; the other in which both cytokines were given concurrently for 5 days only (FL, 200 μg/kg; G-CSF, 100 μg/kg). Both schemes yielded much higher progenitor mobilization levels (peak levels of colony-forming cells [CFSs] 41,000 to 95,000/mL blood) than observed with either FL (CFC 4,600 to 7,300/mL) or G-CSF (8,405 ± 3,024/mL) used alone at the same doses. Furthermore, there was a progressive and significant expansion of progenitors in vitro during 2 weeks in suspension cultures of mononuclear cells or of CD34+ cells only in the animal with the combined treatment. Likewise, substantial mobilization of osteoclastogenic progenitors was documented only with the combined treatment. Given the functional properties of FL, its synergistic mobilization with G-CSF, and its anticipated good tolerance (because of the absence of an effect on mast cell activation), a clinical use is projected for this cytokine in peripheral blood transplantation settings, as well as in experiments with ex vivo gene transfer.

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Quantitative in vivo assay of human granulocyte colony-stimulating factor using cyclophosphamide-induced neutropenic mice

Blood ◽

10.1182/blood.v75.6.1228.1228 ◽

1990 ◽

Vol 75 (6) ◽

pp. 1228-1233 ◽

Cited By ~ 9

Author(s):

K Hattori ◽

K Shimizu ◽

M Takahashi ◽

M Tamura ◽

M Oheda ◽

...

Keyword(s):

Linear Relationship ◽

Neutrophil Count ◽

Peripheral Blood ◽

Assay Method ◽

Blood Neutrophil ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

Peripheral Blood Neutrophil ◽

Stimulating Factor

Abstract Administration of human granulocyte colony-stimulating factor (hG-CSF) to mice with cyclophosphamide (CPA)-induced neutropenia for 4 consecutive days from the day after the CPA dosing (100 mg/kg) resulted in a dose-dependent increase in the peripheral blood neutrophil count 6 hours after the final hG-CSF injection. Within the hG-CSF dose range of 0.1 to 10 micrograms per mouse per day, there was a strong linear relationship (r greater than .9) between the logarithm of the dose and the peripheral blood neutrophil count in the treated mice. Using the same hG-CSF preparation, 38 experiments indicated that the regression lines are highly reproducible. Such an association never occurred with intact mice, and 100 mg/kg of CPA induced the highest response to hG- CSF. This linear relationship between the two variables allows us to determine the biologic potency of a test hG-CSF preparation relative to a reference standard using a parallel line assay, with a coefficient of precision of around .2. When assayed by this bioassay procedure, which we have termed CPA-mouse assay, natural hG-CSF and recombinant hG-CSF (produced by Chinese hamster ovary cells) were nearly equipotent in specific biologic activity. These results confirm the CPA-mouse assay as an especially useful assay method for quantifying the in vivo activity of hG-CSF.

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Granulocyte colony-stimulating factor induces hFc gamma RI (CD64 antigen)-positive neutrophils via an effect on myeloid precursor cells

Blood ◽

10.1182/blood.v81.6.1457.1457 ◽

1993 ◽

Vol 81 (6) ◽

pp. 1457-1464 ◽

Cited By ~ 49

Author(s):

JM Kerst ◽

JG van de Winkel ◽

AH Evans ◽

M de Haas ◽

IC Slaper-Cortenbach ◽

...

Keyword(s):

Peripheral Blood ◽

Precursor Cells ◽

Polymorphonuclear Neutrophil ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

Final Maturation ◽

Vitro Effect ◽

Stimulating Factor

Abstract In this study we have examined hFc gamma RI expression during myelopoiesis. Normal bone marrow (BM) cells were found to express hFc gamma RI up to the metamyelocyte stage. A different Fc gamma RI expression pattern was observed in an in vitro model of myelopoiesis. Purified CD34-positive BM cells, cultured for 12 to 14 days with granulocyte colony-stimulating factor (G-CSF), differentiate into a population of mature granulocytic cells. In these cultures, in which hFc gamma RI was virtually absent on the initial CD34-positive BM cells, hFc gamma RI was strongly induced by G-CSF after only 5 days. During final maturation the cells remained hFc gamma RI positive. This expression was confirmed functionally by antibody-sensitized erythrocytes (EA)-rosette assays. Moreover, the mature myeloid cells were found to express mRNA encoding for hFc gamma RI, whereas reverse- transcriptase polymerase chain reaction analysis showed that both hFc gamma RIA and hFc gamma RIB genes were expressed. In contrast, on peripheral blood (PB) polymorphonuclear neutrophil leukocytes (PMN) the in vitro effect of G-CSF as to hFc gamma RI induction was limited. Therefore, we conclude that, with respect to hFc gamma RI expression on PMN, G-CSF acts on myeloid precursor cells rather than on mature cells. This conclusion could be strengthened by in vivo administration of a single dose of G-CSF to a healthy volunteer. After a 12-hour lag time, hFc gamma RI expressing PMNs were detected in the peripheral blood. This study shows that hFc gamma RI is an early myeloid differentiation marker that is lost during normal final maturation. However, committed myeloid progenitor cells can be strongly induced by G-CSF to express hFc gamma RI, ultimately resulting in mature granulocytic cells expressing the high-affinity receptor for IgG. This expression may have important consequences for the functional capacity of these cells.

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Pharmacologic doses of granulocyte colony-stimulating factor affect cytokine production by lymphocytes in vitro and in vivo

Blood ◽

10.1182/blood.v95.7.2269.007k37_2269_2274 ◽

2000 ◽

Vol 95 (7) ◽

pp. 2269-2274 ◽

Cited By ~ 2

Author(s):

Elaine M. Sloand ◽

Sonnie Kim ◽

Jaroslaw P. Maciejewski ◽

Fritz Van Rhee ◽

Aniruddho Chaudhuri ◽

...

Keyword(s):

Peripheral Blood ◽

Mononuclear Cells ◽

Th2 Cells ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

Cd4 Cells ◽

Ifn Γ ◽

Stimulating Factor

Peripheral blood stem cell (PBSC) transplantation is successful in improving engraftment without increasing acute graft-versus-host disease (GVHD), despite much larger numbers of T cells in unmanipulated PBSCs than in bone marrow grafts. In mouse models and retrospective human studies, granulocyte colony-stimulating factor (G-CSF) therapy has been associated with less acute GVHD. We studied the effect of G-CSF on interferon (IFN)-γ and IL-4 expression in CD4+lymphocytes. CD4+ cells co-cultivated with G-CSF and stimulated with PHA or CD3 monoclonal antibodies showed significant decreases in IFN-γ and increases in IL-4 expression (n = 13;P < .01). G-CSF appeared to have a direct effect on CD4+ cells independent of monocytes present in the culture because purified CD4+ cells exposed to G-CSF, washed, and cocultivated with untreated monocytes demonstrated similar changes in IFN-γ and IL-4 expression, whereas untreated CD4+ cells cocultured with G-CSF–stimulated monocytes behaved as controls. We then studied peripheral blood mononuclear cells (PBMCs) from G-CSF–mobilized PBSC donors. When their PBMCs were cultured with PHA or CD3 monoclonal antibody, the percent of IFN-γ–expressing cells decreased by a mean of 55% and 42%, respectively, whereas the percent of IL-4–containing cells increased by a mean of 39% and 58%, respectively, following G-CSF stimulation. Increased apoptosis of IFN-γ–producing CD4+ cells was not responsible for the shift in TH1/TH2 subsets. G-CSF-R mRNA was present in both CD4+ and CD8+ cells. These results suggest that G-CSF decreases IFN-γ and increases IL-4 production in vitro and in vivo and likely modulates a balance between TH1 and TH2 cells, an effect that may be important in PBSC transplantation.

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Stem cell factor enhances in vivo effects of granulocyte colony stimulating factor for stimulating mobilization of peripheral blood progenitor cells

Stem Cells ◽

10.1002/stem.5530110807 ◽

1996 ◽

Vol 11 (S2) ◽

pp. 36-41 ◽

Cited By ~ 14

Author(s):

Ian K. Mcniece ◽

Robert A. Briddell ◽

Cynthia A. Hartley ◽

Kent A. Smith ◽

Robert G. Andrews

Keyword(s):

Stem Cell ◽

Progenitor Cells ◽

Peripheral Blood ◽

Stem Cell Factor ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

Peripheral Blood Progenitor Cells ◽

In Vivo Effects ◽

Stimulating Factor

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A Specific Stimulator of Granulocyte Colony-Stimulating Factor Accelerates Recovery From Cyclophosphamide-Induced Neutropenia in the Mouse

Blood ◽

10.1182/blood.v90.2.795.795_795_802 ◽

1997 ◽

Vol 90 (2) ◽

pp. 795-802

Author(s):

Jay S. Fine ◽

Xiao-Yan Cai ◽

Luminita Justice ◽

Carl P. Gommoll ◽

Linda D. Hamilton ◽

...

Keyword(s):

Peripheral Blood ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

Small Molecular Weight ◽

Peripheral Blood Mononuclear ◽

Stimulating Factor

We have identified a small molecular weight compound, SCH 14988, which specifically stimulates in vitro granulocyte-colony stimulating factor (G-CSF ) production from activated human peripheral blood mononuclear cells and monocytes but not other cytokines or CSFs with hematoregulatory activity. In vivo administration of SCH 14988 to mice rendered neutropenic by cyclophosphamide treatment resulted in the accelerated recovery of the peripheral neutrophil compartment. This activity correlated with increased in vivo G-CSF levels and stimulation of marrow granulopoiesis, and was comparable to that of exogenously administered recombinant human G-CSF. No alterations to other leukocyte populations in peripheral blood, spleen, or the peritoneal cavity were observed. These findings suggest that SCH 14988 may be clinically useful to enhance neutrophil granulopoiesis, as well as to study the mechanisms involved in G-CSF gene regulation.

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