scholarly journals Correlation between the stimulation of human neutrophil function by monoclonal antibody and by colony-stimulating factor

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 738-741 ◽  
Author(s):  
MA Vadas ◽  
C Clarke ◽  
NA Nicola ◽  
AF Lopez
Keyword(s):  
Monoclonal Antibody ◽  
Neutrophil Function ◽  
Common Mechanism ◽  
Human Neutrophils ◽  
Target Cells ◽  
Colony Stimulating Factor ◽  
Positive Correlation ◽  
Stimulating Factor ◽  
Stimulation Of

Abstract Purified human neutrophils from 48 individuals were tested for their capacity to kill antibody-coated target cells in vitro in the absence or presence of stimulating agents. The agents used to stimulate cytotoxic capacity were the monoclonal antibody (MAb) WEM-G1, colony- stimulating factor (CSF-alpha), or mononuclear cell supernatant (MNC- SN). There existed an heterogeneity among the neutrophils of different individuals in the capacity to kill target cells both in the unstimulated (“resting”) or the stimulated state. A positive correlation was found between the ability of neutrophils to kill in the “resting” state and their capacity to be stimulated by MAb WEM-G1, CSF- alpha, or MNC-SN. Furthermore, a strong positive correlation in the ability of neutrophils to be stimulated by the MAb WEM-G1 and either CSF-alpha (r = .76) or MNC-SN (r = .68), as well as between CSF-alpha and MNC-SN (r = .79) was demonstrated. No correlation was seen, however, between stimulation of neutrophil function in vitro and total blood leukocyte counts, neutrophil counts, monocyte counts, or intensity of binding of MAb WEM-G1. The observation that neutrophils respond to a similar extent to different types of stimulators, -such as cytokines (CSF-alpha and MNC-SN) and MAb, suggests that these two factors may be operating through a common mechanism and the degree of stimulation may reflect an intrinsic responsiveness of neutrophils that differs among individuals. Our results also suggest a potential clinical use of WEM-G1 in measuring neutrophil functional capacity in vitro and predicting the capacity to respond to CSF-like cytokines.

scholarly journals Correlation between the stimulation of human neutrophil function by monoclonal antibody and by colony-stimulating factor

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 738-741
Author(s):  
MA Vadas ◽  
C Clarke ◽  
NA Nicola ◽  
AF Lopez
Keyword(s):  
Monoclonal Antibody ◽  
Neutrophil Function ◽  
Common Mechanism ◽  
Human Neutrophils ◽  
Target Cells ◽  
Colony Stimulating Factor ◽  
Positive Correlation ◽  
Stimulating Factor ◽  
Stimulation Of

Purified human neutrophils from 48 individuals were tested for their capacity to kill antibody-coated target cells in vitro in the absence or presence of stimulating agents. The agents used to stimulate cytotoxic capacity were the monoclonal antibody (MAb) WEM-G1, colony- stimulating factor (CSF-alpha), or mononuclear cell supernatant (MNC- SN). There existed an heterogeneity among the neutrophils of different individuals in the capacity to kill target cells both in the unstimulated (“resting”) or the stimulated state. A positive correlation was found between the ability of neutrophils to kill in the “resting” state and their capacity to be stimulated by MAb WEM-G1, CSF- alpha, or MNC-SN. Furthermore, a strong positive correlation in the ability of neutrophils to be stimulated by the MAb WEM-G1 and either CSF-alpha (r = .76) or MNC-SN (r = .68), as well as between CSF-alpha and MNC-SN (r = .79) was demonstrated. No correlation was seen, however, between stimulation of neutrophil function in vitro and total blood leukocyte counts, neutrophil counts, monocyte counts, or intensity of binding of MAb WEM-G1. The observation that neutrophils respond to a similar extent to different types of stimulators, -such as cytokines (CSF-alpha and MNC-SN) and MAb, suggests that these two factors may be operating through a common mechanism and the degree of stimulation may reflect an intrinsic responsiveness of neutrophils that differs among individuals. Our results also suggest a potential clinical use of WEM-G1 in measuring neutrophil functional capacity in vitro and predicting the capacity to respond to CSF-like cytokines.

Receptor expression and oxidase activity in human neutrophils: Regulation by granulocyte-macrophage colony-stimulating factor and dependence upon protein biosynthesis

1990 ◽  
Vol 10 (4) ◽  
pp. 393-401 ◽  
Author(s):  
Steven W. Edwards ◽  
Fiona Watson ◽  
Ronald MacLeod ◽  
John Davies
Keyword(s):  
De Novo ◽  
Protein Biosynthesis ◽  
Neutrophil Function ◽  
Membrane Receptors ◽  
Receptor Expression ◽  
Human Neutrophils ◽  
Colony Stimulating Factor ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Incubation of human bloodstream neutrophils with 50 u/ml recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF) “primed” the respiratory burst (as assessed by fMet-Leu-Phe stimulated luminol-dependent chemiluminescence) and resulted in a rapid (within 15 min) upregulation of expression of CD11b and CD18 (as measured by FACS analysis). This rapid “priming” and modulation of receptor expression was not inhibited by cycloheximide and hence appeared to be independent of de novo protein biosynthesis. When neutrophils were incubated for up to 5 h in culture, the fluorescence distributions of CD11b and CD18 declined indicating the loss of expression of these receptors as the neutrophils aged, but in rGM-CSF treated suspensions receptor expression was maintained. When neutrophils were incubated in the presence of cycloheximide, they progressively lost their ability to generate reactive oxidants in response to fMet-Leu-Phe so that by 5 h incubation with this inhibitor they could only generate about 25% of the oxidative response stimulated in untreated cells, and the expression of CD16 and CD18 was grossly impaired. Similar effects were observed in rGM-CSF treated suspensions except that cycloheximide required longer incubation times (typically 4–5 h) before impairment of function or receptor expression occurred. These data show that de novo protein biosynthesis is required for both the maintenance of neutrophil function and also for the continued expression of some plasma membrane receptors.

scholarly journals Inhibition of interleukin 3 and colony-stimulating factor 1-stimulated marrow cell proliferation by pertussis toxin

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1187-1195 ◽  
Author(s):  
YX He ◽  
E Hewlett ◽  
D Temeles ◽  
P Quesenberry
Keyword(s):  
Bone Marrow ◽  
G Proteins ◽  
Pertussis Toxin ◽  
Cell Surface Receptor ◽  
Target Cells ◽  
Colony Stimulating Factor ◽  
Murine Bone Marrow ◽  
Interleukin 3 ◽  
Stimulating Factor ◽  
Stimulation Of

Abstract Pertussis toxin (PT) catalyzes the ADP-ribosylation of several guanine nucleotide-binding (G) proteins that are involved in the transduction of cell surface receptor-mediated signals. Involvement of such G- proteins in regulation of hematopoiesis by two growth factors, colony- stimulating factor-1 (CSF-1) and interleukin 3 (IL 3), was investigated using pertussis toxin. Continuous or pulse exposure of murine bone marrow cells to pertussis toxin inhibited CSF-1 or IL 3-induced colony formation by approximately 50%. Pertussis toxin inhibition was also demonstrated against partially separated marrow from 5-fluorouracil- treated mice. The toxin effect was blocked by heating (95 degrees C for 30 minutes), by antitoxin antibody and was not associated with increased cAMP levels in target cells. In experiments with murine marrow, toxin-mediated inhibition appeared to involve predominantly the macrophage lineage. IL 3 stimulation of proliferation of the murine marrow-derived factor-dependent cell line FDC-P1, as measured by 3H-TdR incorporation, and CSF-1 stimulation of pure populations of murine bone marrow derived macrophages, as measured by DNA content and cell number, was also inhibited. Analysis of the effects of pertussis toxin on the growth of single cells stimulated by IL 3 demonstrated that this inhibition involved a decreased growth rate rather than a toxic ablation of cells. Phorbol myristate acetate (PMA) stimulated FDC-P1 cells and was able to abrogate the PT inhibition of IL 3 stimulation of these cells, suggesting but not establishing that IL 3 may mediate its proliferative effects through activating protein kinase C.

scholarly journals Target cells for granulocyte colony-stimulating factor, interleukin-3, and interleukin-5 in differentiation pathways of neutrophils and eosinophils

Blood ◽  
1990 ◽  
Vol 76 (10) ◽  
pp. 1956-1961 ◽  
Author(s):  
H Ema ◽  
T Suda ◽  
K Nagayoshi ◽  
Y Miura ◽  
CI Civin ◽  
...  
Keyword(s):  
Bone Marrow Cells ◽  
Early Stage ◽  
Target Cells ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Interleukin 3 ◽  
Cd34 Antigen ◽  
Cd34 Cells ◽  
Stimulating Factor

Abstract To study the relationship between hematopoietic factors and their responsive hematopoietic progenitors in the differentiation process, both purified factors and enriched progenitors are required. We isolated total CD34+ cells, CD34+,CD33+ cells, and CD34+,CD33- cells individually from normal human bone marrow cells by fluorescence- activated cell sorter (FACS), and examined the effects of granulocyte colony-stimulating factor (G-CSF), interleukin-3 (IL-3), and IL-5 on in vitro colony formation of these cells. CD34+,CD33+ cells formed granulocyte colonies in the presence of G-CSF. Both CD34+,CD33+ cells and CD34+,CD33- cells formed granulocyte/macrophage colonies in the presence of IL-3. Eosinophil (Eo) colonies were only formed by CD34+,CD33- cells in response to IL-3, but scarcely formed by CD34+ cells in the presence of IL-5. We performed the two-step cultures consisting of the primary liquid culture for 6 days and the secondary methylcellulose culture, and serially examined changes in phenotypes of ,he cells cultured in the primary culture. CD34-,CD33+ cells derived from CD34+,CD33+ cells by preincubation with G-CSF or IL-3 formed Eo colonies in the presence of IL-5 but not IL-3. CD34-,CD33+ cells derived from CD34+,CD33- cells by preincubation with IL-3 also formed Eo colonies by support of IL-5 as well as IL-3. Both CD34+ cells gradually lost the CD34 antigen by day 6 of incubation with G-CSF or IL- 3. Loss of this antigen was well-correlated with acquisition of susceptibility to IL-5. It was concluded that G-CSF supported the neutrophil differentiation of committed colony-forming cells, IL-3 supported that of both committed and multipotent colony-forming cells. G-CSF and IL-3 also supported the early stage of E. differentiation; IL- 5 supported the late stage of that.

A potential therapeutic role for small nonpeptidyl compounds that mimic human granulocyte colony-stimulating factor

Blood ◽  
2004 ◽  
Vol 103 (3) ◽  
pp. 836-842 ◽  
Author(s):  
Kouji Kusano ◽  
Shinji Ebara ◽  
Koichi Tachibana ◽  
Tadahiro Nishimura ◽  
Susumu Sato ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Interleukin 2 ◽  
Immune Defense ◽  
Mitogen Activated Protein Kinase ◽  
Human Neutrophils ◽  
Blood Neutrophil ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Stimulating Factor

AbstractGranulocyte colony-stimulating factor (G-CSF) stimulates the proliferation of bone marrow granulocytic progenitor cells and promotes their differentiation into granulocytes. G-CSF is therefore an important component of immune defense against pathogenic microorganisms: recombinant human G-CSF (rhG-CSF) is used to treat patients with a variety of neutropenias. In the present study, we screened approximately 10 000 small nonpeptidyl compounds and found 3 small compounds that mimic G-CSF in several in vitro and in vivo assays. These compounds induced G-CSF–dependent proliferation, but had no effect on interleukin-3–dependent, interleukin-2–dependent, interleukin-10–dependent, thrombopoietin (TPO)–dependent, or erythropoietin (EPO)–dependent proliferation. Each compound induced the phosphorylation of signal transducers and activators of transcription–3 (STAT3) and mitogen-activated protein kinase (MAPK) in a G-CSF–dependent cell line and in human neutrophils. In addition, these compounds induced hematopoietic colony formation from primary rat bone marrow cells in vitro. When subcutaneously injected into normal rats, they caused an increase in peripheral blood neutrophil counts. Furthermore, when they were administered to cyclophosphamide-induced neutropenic rats, blood neutrophil levels increased and remained elevated up to day 8. We therefore suggest that these small nonpeptidyl compounds mimic the activity of G-CSF and may be useful in the treatment of neutropenic patients.

scholarly journals In vivo stimulation of megakaryocytopoiesis by recombinant murine granulocyte-macrophage colony-stimulating factor

Blood ◽  
1990 ◽  
Vol 76 (8) ◽  
pp. 1473-1480
Author(s):  
AM Vannucchi ◽  
A Grossi ◽  
D Rafanelli ◽  
PR Ferrini
Keyword(s):  
Bone Marrow ◽  
Blood Platelets ◽  
Colony Stimulating Factor ◽  
Platelet Production ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Stimulation Of

Murine recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF) was injected in mice, and the effects on bone marrow, splenic megakaryocytes, megakaryocyte precursors (megakaryocyte colony-forming units [CFU-Meg]) were evaluated. In mice injected three times a day for 6 days with 12,000 to 120,000 U rGM-CSF, no significant modification of both platelet levels and mean platelet volume was observed, while there was a twofold increase in blood neutrophils. However, the rate of platelet production, as assessed by the measurement of 75selenomethionine incorporation into blood platelets, was On the contrary, administration of up to 384,000 U rGM-CSF two times a day for 2 days, as for a typical “thrombopoietin assay,” failed to modify platelet production. A significant dose-related increase in the number of splenic megakaryocytes occurred in mice receiving 60,000 to 120,000 U rGM-CSF, while a slight increase in the number of bone marrow megakaryocytes was observed in mice injected with 120,000 U rGM-CSF. The proportion of bone marrow megakaryocytes with a size less than 18 microns and greater than 35 microns resulted significantly higher in mice receiving rGM-CSF in comparison with controls; an increase in the percentage of splenic megakaryocytes greater than 35 microns was also observed. A statistically significant increase in the total spleen content of CFU-Meg was observed after administration of 90,000 and 120,000 U rGM-CSF three times a day for 6 days, while no effect on bone marrow CFU-Meg was recorded, irrespective of the dose delivered. Finally, 24 hours after a single intravenous injection of rGM-CSF, there was a significant increase in the proportion of CFU-Meg in S- phase, with the splenic progenitors being more sensitive than bone marrow-derived CFU-Meg. These data indicate that rGM-CSF has in vivo megakaryocyte stimulatory activity, and are consistent with previous in vitro observations. However, an effective stimulation of megakaryocytopoiesis in vivo, bringing about an increase in the levels of blood platelets, may require interaction of rGM-CSF with other cytokines.

scholarly journals Granulocyte colony-stimulating factor stimulates human mature neutrophilic granulocytes to produce interferon-alpha

Blood ◽  
1990 ◽  
Vol 75 (1) ◽  
pp. 17-19 ◽  
Author(s):  
N Shirafuji ◽  
S Matsuda ◽  
H Ogura ◽  
K Tani ◽  
H Kodo ◽  
...  
Keyword(s):  
Interferon Alpha ◽  
Messenger Rna ◽  
Mononuclear Cells ◽  
Human Neutrophils ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Glycoprotein Hormone ◽  
Ifn Alpha ◽  
Stimulating Factor

Abstract Granulocyte colony-stimulating factor (G-CSF) is a glycoprotein hormone that specifically stimulates both production and functional activation of neutrophils, while interferon-alpha (IFN-alpha) is known to suppress myelopoiesis, including neutrophil production in vivo and in vitro. On a possibility that IFN-alpha may operate as one of the inhibitory feedback factors in neutropoiesis, we examined whether neutrophils produce IFN-alpha in response to G-CSF. Northern blot analysis showed that messenger RNA (mRNA) for human IFN-alpha 1 became detectable time- dependently in highly purified human neutrophils incubated with purified recombinant human G-CSF (rhG-CSF). But such transcription was not observed either in neutrophils incubated with other neutrophil activators, such as formyl-methionyl-leucyl-phenylalanine (fMLP) or lipopolysaccharides (LPS), or in blood mononuclear cells incubated with rhG-CSF. In addition, radioimmunoassay for human IFN-alpha showed that its levels in culture medium of the rhG-CSF-treated neutrophils rose markedly (up to approximately 100 IU/mL/1 x 10(7) cells) in a time- dependent way, compared with those of nonstimulated neutrophils. These findings suggest that the G-CSF/IFN-alpha system may participate in the feedback regulatory loop of neutropoiesis.

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