Granulocyte - macrophage colony stimulating factor and interleukin-8 in the reproductive tract of ewes following oestrus and mating

2007 ◽  
Vol 19 (4) ◽  
pp. 585 ◽  
Author(s):  
Jennifer L. Scott ◽  
Natkunam Ketheesan ◽  
Phillip M. Summers
Keyword(s):  
Reproductive Tract ◽  
Staining Intensity ◽  
Female Reproductive Tract ◽  
Enzyme Linked Immunosorbent Assay ◽  
Colony Stimulating Factor ◽  
Tissue Samples ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Cytokines produced in the female reproductive tract after mating may enhance reproductive success. The present study investigated the distribution of granulocyte–macrophage colony stimulating factor (GM-CSF) and interleukin (IL)-8 in tissues and luminal secretions from different sites in the reproductive tract of the ewe following oestrus and after natural mating. Fifteen ewes were mated with a ram for 1 h and their reproductive tracts collected 3, 6, 18, 24 or 48 h later. Another 15 ewes were used as oestrous controls. Luminal secretions and tissue samples were collected from seven sites in each reproductive tract. Secretions were analysed by enzyme-linked immunosorbent assay and tissues were stained immunohistochemically using anti-sheep GM-CSF and anti-sheep IL-8 antibodies. Both cytokines were found in luminal and glandular endometrial epithelium and, to a lesser extent, in cervical epithelium; neither was found in the vaginal epithelium. Twice as many (P < 0.05) luminal samples from mated ewes than non-mated ewes were positive for GM-CSF. The vaginal lumen contained significantly higher (P < 0.01) concentrations of IL-8 compared with other sites, irrespective of mating status. Significant differences (P < 0.05) were found in staining intensity of GM-CSF and IL-8 from different sites. Production of GM-CSF and IL-8 by reproductive tissues is likely to contribute to leucocyte infiltration into the ovine reproductive tract.

scholarly journals Induction of anti-recombinant human granulocyte-macrophage colony- stimulating factor (Escherichia coli-derived) antibodies and clinical effects in nonimmunocompromised patients

Blood ◽  
1994 ◽  
Vol 84 (12) ◽  
pp. 4078-4087 ◽  
Author(s):  
P Ragnhammar ◽  
HJ Friesen ◽  
JE Frodin ◽  
AK Lefvert ◽  
M Hassan ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibody Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Clinical Effects ◽  
Immunocompromised Patients ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

The pharmacokinetics of recombinant human granulocyte-macrophage colony- stimulating factor (rhGM-CSF), induction of anti-GM-CSF antibodies, and clinical effects related to the induction of the antibodies were analyzed in patients with metastatic colorectal carcinoma (CRC) who were not on chemotherapy (n = 20, nonimmunocompromised patients). rhGM- CSF (250 micrograms/m2/d; Escherichia coli-derived) was administered subcutaneously for 10 days every month for 4 months. Eight patients with multiple myeloma (MM) on intensive chemotherapy followed by rhGM- CSF treatment were also included (immunocompromised patients). After a single injection of GM-CSF at the first cycle in CRC patients, the maximum calculated concentration (Cmax) was 5.24 +/- 0.56 ng/mL; the half life (T1/2) was 2.91 +/- 0.8 hours; and the area under the concentration curve (AUC) was 30.86 +/- 6.03 hours x ng/mL (mean +/- SE). No anti-GM-CSF antibodies were detected. During the subsequent cycles, 95% of the CRC patients developed anti-GM-CSF IgG antibodies, which significantly altered the pharmacokinetics of rhGM-CSF at the third and fourth cycles with decreased Cmax (2.87 +/- 0.57 ng/mL; P < .05), T1/2 (1.57 +/- 0.2 hours; P < .05), and AUC (14.90 +/- 4.10 hours x ng/mL; P < .005). The presence of anti-GM-CSF antibodies significantly reduced the GM-CSF-induced enhancement of granulocytes, and there was a clear tendency for a decreased increment of monocytes. Antibodies diminished systemic side effects of rhGM-CSF. Only 1 of 8 MM patients showed a very low anti-GM-CSF antibody titer after GM-CSF therapy, as shown by enzyme-linked immunosorbent assay and Western blot. Therefore, in nonimmunocompromised patients, exogenous nonglycosylated GM-CSF induced an anti-GM-CSF IgG antibody response in practically all patients, which seemed to be of clinical significance. In immunocompromised patients, virtually no significant antibody response was shown.

scholarly journals Identification of functionally distinct domains of human granulocyte- macrophage colony-stimulating factor using monoclonal antibodies

Blood ◽  
1991 ◽  
Vol 77 (5) ◽  
pp. 1033-1043 ◽  
Author(s):  
Y Kanakura ◽  
SA Cannistra ◽  
CB Brown ◽  
M Nakamura ◽  
GF Seelig ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Neutralizing Antibodies ◽  
Solid Phase ◽  
Receptor Interaction ◽  
Enzyme Linked Immunosorbent Assay ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Abstract Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a glycoprotein that is required for the survival, growth, and differentiation of hematopoietic progenitor cells. Although the primary structure of GM-CSF is known from cDNA cloning, the relationship between structure and function of GM-CSF is not fully understood. Fifteen different monoclonal antibodies (MoAbs) to human GM-CSF were generated to map immunologically distinct areas of the molecule. Each of the MoAbs was biotinylated and shown by enzyme-linked immunosorbent assay to bind to recombinant GM-CSF that had been affixed to a solid phase. Each of the 15 unconjugated MoAbs was then used to compete with each biotinylated MoAb for binding to GM-CSF. These cross-blocking studies identified eight distinct epitopes of native GM-CSF. Seven of these epitopes were also present in denatured GM-CSF by Western blotting, and four of the epitopes were at least partially conserved on GM-CSF that was reduced in beta-mercaptoethanol. MoAbs to four of eight epitopes neutralized both recombinant (glycosylated and nonglycosylated) and natural human GM-CSF in a GM colony-forming unit (CFU-GM) assay and blocked GM-CSF-induced activation of neutrophils. For most of the antibodies there was a good correlation between neutralizing activity and the capacity to block binding of 125I-GM-CSF to neutrophils or blasts. Non-neutralizing antibodies to one epitope partially blocked binding of 125I-GM-CSF to neutrophils. None of the MoAbs neutralized interleukin-3, G-CSF, or M-CSF. The locations of seven of the epitopes could be partially mapped with regard to the amino acid structure by determining reactivity to GM-CSF synthetic peptides or to human-mouse chimeric GM-CSFs. The neutralizing antibodies were found to map to amino acids 40–77, 78–94, or 110–127. Thus, these MoAbs are useful to identify functional domains of GM-CSF and in identifying regions that are likely to be involved in receptor interaction.

167 THE SUPPLEMENTATION OF GRANULOCYTE-MACROPHAGE COLONY STIMULATING FACTOR (GM-CSF) PROMOTES THE DEVELOPMENT OF NUCLEAR TRANSFERRED BOVINE EMBRYOS

2006 ◽  
Vol 18 (2) ◽  
pp. 191
Author(s):  
D. H. Kim ◽  
S. W. Kim ◽  
B. C. Yang ◽  
G. S. Im ◽  
H. S. Park ◽  
...  
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Female Reproductive Tract ◽  
Apoptotic Cells ◽  
Colony Stimulating Factor ◽  
Bovine Embryos ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Granulocyte-macrophage colony stimulating factor (GM-CSF) is secreted by epithelial cells lining the female reproductive tract in mice and several other species. GM-CSF receptors are present in the fertilized oocyte and in all subsequent stages of development, and in blastocysts it is expressed in both inner cell mass and trophectoderm cells. Recent studies suggest that GM-CSF can act as a survival factor for the developing embryo. The purpose of this study was to examine the effect of GM-CSF, as a medium supplement, on the development of nuclear-transferred bovine embryos. Oocytes were enucleated after in vitro maturation in TCM-199 supplemented with 10% fetal bovine serum (FBS), 1 mg/mL FSH, and 1 mg/mL estradiol-17� for 20 h. Enucleated oocytes were fused with bovine ear skin fibroblast cells by a DC pulse of 25 V/150 mm for 20 ms in Zimmerman cell fusion medium. For activation, reconstructed embryos were exposed to 10 mM Ca-ionophore for 5 min, followed by 2 mM 6-dimethylaminopurine for 3 h. NT embryos were subsequently cultured in CR2 medium without or with 10 ng/mL recombinant porcine GM-CSF at 39.0�C in 5% O2, 5% CO2 and 90% N2. After 7 days of culture, blastocyst formation was observed. The number of inner cell mass (ICM) and trophectoderm (TE) cells was examined by differential staining. Apoptotic cells in blastocysts were detected by a terminal deoxynucleotidyl transferase-mediated d-UTP nick-end labeling (TUNEL) assay. Data were analyzed by chi-square and Student's t-test. Addition of GM-CSF to the medium significantly (P < 0.05) increased the proportion of embryos developing to the blastocyst stage (37.6 � 12.0 and 54.7 � 13.9% for control and GM-CSF groups respectively). No differences in the total cell number and the ratio of ICM to total cells were detected between the control group (125.4 � 35.7 and 38.5 � 9.7%) and the GM-CSF group (123.8 � 35.1 and 34.2 � 13.1%). The mean proportion of apoptotic cells in blastocysts was not different between the control (5.4 � 5.4%) and the GM-CSF (5.3 � 3.9%) group. Our results showed the beneficial effect of GM-CSF on the development of NT bovine embryos. These results suggest that GM-CSF might be a useful molecule for increasing development of NT bovine embryos. Further studies are necessary to verify the mechanism of GM-CSF on the development of bovine NT embryos.

288. The post-insemination inflammatory response in the ewe

2005 ◽  
Vol 17 (9) ◽  
pp. 121 ◽  
Author(s):  
J. L. Scott ◽  
N. Ketheesan ◽  
P. M. Summers
Keyword(s):  
Inflammatory Response ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Enzyme Linked Immunosorbent Assay ◽  
Tissue Samples ◽  
Gm Csf ◽  
Reproductive Tissues ◽  
Cell Counts ◽  
Macrophage Colony ◽  
Uterine Body

Insemination causes an inflammatory response in the female reproductive tract of many species. The cytokine/leukocyte network initiated during this reaction is believed to enhance reproductive success.1 This study investigated the post-insemination inflammatory response in the ewe. Fifteen nonparous ewes were mated with the same ram for 1 h and their reproductive tracts were collected 3, 6, 18, 24 or 48 h later. Another fifteen ewes were used as controls. Tissue samples and luminal mucus were collected from 10 sites in each reproductive tract and stained with haematoxylin and eosin, Diffquik and immunohistochemically using a monoclonal CD68 antibody to quantify neutrophils, eosinophils and macrophages. Presence of interleukin-8 (IL-8) and granulocyte-macrophage colony-stimulating factor (GM-CSF) was investigated using immunohistochemistry and enzyme-linked immunosorbent assay. Neutrophils and macrophages increased in reproductive tissues following insemination. Mean cell counts in 1.5-mm2 tissue of mated (M) and control (C) ewes demonstrated a peak in neutrophils at 6–18 h post-insemination with significant differences (P < 0.05) between mated and controls in the posterior cervix (M = 23.7; C = 4.1) and uterine body (M = 34.5; C = 11.5). Macrophages peaked at 18–24 h with significant differences (P < 0.05) between mated and controls in the vagina (M=13.4; C = 4.6), posterior cervix (M = 10.4; C = 2.7), mid-cervix (M = 8.5; C = 3.0) and ipsilateral mid-uterine horn (M = 14.2; C = 7.9). Neutrophils increased in the lumen of the cervix and uterine body following insemination but macrophage numbers did not change. Insemination did not affect eosinophils. IL-8 and GM-CSF were detected in endometrial epithelial cells in mated and non-mated ewes. Highest concentrations of IL-8 were found in vaginal mucus. Small quantities of GM-CSF were detected in occasional mucus samples. No difference between mated and non-mated ewes was demonstrated for either cytokine. In conclusion, the post-insemination inflammatory reaction in the ewe involves an increase in neutrophils and macrophages in reproductive tissues, with neutrophils crossing the epithelium into the lumen. There was no apparent increase in IL-8 or GM-CSF in response to insemination. (1)Robertson SA et al. (1997) American Journal of Reproductive Immunology 37, 438–442.

Measurement of human granulocyte-macrophage colony-stimulating factor (GM-CSF) by enzyme-linked immunosorbent assay

Biotherapy ◽  
1989 ◽  
Vol 1 (3) ◽  
pp. 161-167 ◽  
Author(s):  
Fusayuki Omori ◽  
Seiichi Okamura ◽  
Shin Hayashi ◽  
Shigeru Yamaga ◽  
Yuichi Hirota ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

scholarly journals Identification of functionally distinct domains of human granulocyte- macrophage colony-stimulating factor using monoclonal antibodies

Blood ◽  
1991 ◽  
Vol 77 (5) ◽  
pp. 1033-1043
Author(s):  
Y Kanakura ◽  
SA Cannistra ◽  
CB Brown ◽  
M Nakamura ◽  
GF Seelig ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Neutralizing Antibodies ◽  
Solid Phase ◽  
Receptor Interaction ◽  
Enzyme Linked Immunosorbent Assay ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a glycoprotein that is required for the survival, growth, and differentiation of hematopoietic progenitor cells. Although the primary structure of GM-CSF is known from cDNA cloning, the relationship between structure and function of GM-CSF is not fully understood. Fifteen different monoclonal antibodies (MoAbs) to human GM-CSF were generated to map immunologically distinct areas of the molecule. Each of the MoAbs was biotinylated and shown by enzyme-linked immunosorbent assay to bind to recombinant GM-CSF that had been affixed to a solid phase. Each of the 15 unconjugated MoAbs was then used to compete with each biotinylated MoAb for binding to GM-CSF. These cross-blocking studies identified eight distinct epitopes of native GM-CSF. Seven of these epitopes were also present in denatured GM-CSF by Western blotting, and four of the epitopes were at least partially conserved on GM-CSF that was reduced in beta-mercaptoethanol. MoAbs to four of eight epitopes neutralized both recombinant (glycosylated and nonglycosylated) and natural human GM-CSF in a GM colony-forming unit (CFU-GM) assay and blocked GM-CSF-induced activation of neutrophils. For most of the antibodies there was a good correlation between neutralizing activity and the capacity to block binding of 125I-GM-CSF to neutrophils or blasts. Non-neutralizing antibodies to one epitope partially blocked binding of 125I-GM-CSF to neutrophils. None of the MoAbs neutralized interleukin-3, G-CSF, or M-CSF. The locations of seven of the epitopes could be partially mapped with regard to the amino acid structure by determining reactivity to GM-CSF synthetic peptides or to human-mouse chimeric GM-CSFs. The neutralizing antibodies were found to map to amino acids 40–77, 78–94, or 110–127. Thus, these MoAbs are useful to identify functional domains of GM-CSF and in identifying regions that are likely to be involved in receptor interaction.

scholarly journals Cholesterol loading suppresses the atheroinflammatory gene polarization of human macrophages induced by colony stimulating factors

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jani Lappalainen ◽  
Nicolas Yeung ◽  
Su D. Nguyen ◽  
Matti Jauhiainen ◽  
Petri T. Kovanen ◽  
...  
Keyword(s):  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Foam Cells ◽  
Colony Stimulating Factor ◽  
Human Macrophages ◽  
Gm Csf ◽  
Macrophage Subpopulations ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

AbstractIn atherosclerotic lesions, blood-derived monocytes differentiate into distinct macrophage subpopulations, and further into cholesterol-filled foam cells under a complex milieu of cytokines, which also contains macrophage-colony stimulating factor (M-CSF) and granulocyte–macrophage-colony stimulating factor (GM-CSF). Here we generated human macrophages in the presence of either M-CSF or GM-CSF to obtain M-MØ and GM-MØ, respectively. The macrophages were converted into cholesterol-loaded foam cells by incubating them with acetyl-LDL, and their atheroinflammatory gene expression profiles were then assessed. Compared with GM-MØ, the M-MØ expressed higher levels of CD36, SRA1, and ACAT1, and also exhibited a greater ability to take up acetyl-LDL, esterify cholesterol, and become converted to foam cells. M-MØ foam cells expressed higher levels of ABCA1 and ABCG1, and, correspondingly, exhibited higher rates of cholesterol efflux to apoA-I and HDL2. Cholesterol loading of M-MØ strongly suppressed the high baseline expression of CCL2, whereas in GM-MØ the low baseline expression CCL2 remained unchanged during cholesterol loading. The expression of TNFA, IL1B, and CXCL8 were reduced in LPS-activated macrophage foam cells of either subtype. In summary, cholesterol loading converged the CSF-dependent expression of key genes related to intracellular cholesterol balance and inflammation. These findings suggest that transformation of CSF-polarized macrophages into foam cells may reduce their atheroinflammatory potential in atherogenesis.

The Truncated Splice Variant of the Granulocyte-Macrophage-Colony-Stimulating Factor Receptor β- Chain in Peripheral Blood Serves as Severity Biomarker of Respiratory Failure in Newborns

Neonatology ◽  
2021 ◽  
pp. 1-7
Author(s):  
Verena Schulte ◽  
Alexandra Sipol ◽  
Stefan Burdach ◽  
Esther Rieger-Fackeldey
Keyword(s):  
Respiratory Failure ◽  
Peripheral Blood ◽  
Colony Stimulating Factor ◽  
Term Infants ◽  
Respiratory Impairment ◽  
Gm Csf ◽  
A Subunit ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

<b><i>Background:</i></b> The granulocyte-macrophage-colony-stimulating factor (GM-CSF) plays an important role in surfactant homeostasis. β<sub>C</sub> is a subunit of the GM-CSF receptor (GM-CSF-R), and its activation mediates surfactant catabolism in the lung. β<sub>IT</sub> is a physiological, truncated isoform of β<sub>C</sub> and is known to act as physiological inhibitor of β<sub>C</sub>. <b><i>Objective:</i></b> The aim of this study was to determine the ratio of β<sub>IT</sub> and β<sub>C</sub> in the peripheral blood of newborns and its association with the degree of respiratory failure at birth. <b><i>Methods:</i></b> We conducted a prospective cohort study in newborns with various degrees of respiratory impairment at birth. Respiratory status was assessed by a score ranging from no respiratory impairment (0) to invasive respiratory support (3). β<sub>IT</sub> and β<sub>C</sub> expression were determined in peripheral blood cells by real-time PCR. β<sub>IT</sub> expression, defined as the ratio of β<sub>IT</sub> and β<sub>C</sub>, was correlated with the respiratory score. <b><i>Results:</i></b> β<sub>IT</sub> expression was found in all 59 recruited newborns with a trend toward higher β<sub>IT</sub> in respiratory ill (score 2, 3) newborns than respiratory healthy newborns ([score 0, 1]; <i>p</i> = 0.066). Seriously ill newborns (score 3) had significantly higher β<sub>IT</sub> than healthy newborns ([score 0], <i>p</i> = 0.010). Healthy preterm infants had significantly higher β<sub>IT</sub> expression than healthy term infants (<i>p</i> = 0.019). <b><i>Conclusions:</i></b> β<sub>IT</sub> is expressed in newborns with higher expression in respiratory ill than respiratory healthy newborns. We hypothesize that β<sub>IT</sub> may have a protective effect in postnatal pulmonary adaptation acting as a physiological inhibitor of β<sub>C</sub> and, therefore, maintaining surfactant in respiratory ill newborns.

Sign in / Sign up

Export Citation Format

Share Document