Reconstitution of nerve growth factor receptor function: Transfer of membranes or vesicles enriched in NGF receptor to a mammalian fibroblast cell line

1986 ◽  
Vol 3 ◽  
pp. S38
Author(s):  
Tatsuro Koike
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cell Line ◽  
Growth Factor Receptor ◽  
Fibroblast Cell ◽  
Fibroblast Cell Line ◽  
Receptor Function ◽  
Nerve Growth ◽  
Ngf Receptor ◽  
Function Transfer

scholarly journals Lateral diffusion of nerve growth factor receptor: modulation by ligand-binding and cell-associated factors.

1990 ◽  
Vol 1 (8) ◽  
pp. 605-614 ◽  
Author(s):  
G Venkatakrishnan ◽  
C A McKinnon ◽  
A H Ross ◽  
D E Wolf
Keyword(s):  
Monoclonal Antibody ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cell Line ◽  
Pc12 Cells ◽  
Growth Factor Receptor ◽  
Lateral Diffusion ◽  
Nerve Growth Factor Receptor ◽  
Nerve Growth ◽  
Cos Cells

We compared the properties in human melanoma cell line A875 and rat pheochromocytoma cell line PC12 of nerve growth factor receptor (NGFr). We also analyzed NGFr and a truncated NGFR lacking the cytoplasmic domain, which were transiently expressed in COS cells. The full-length NGFR expressed in COS cells bound nerve growth factor (NGF) with positive cooperativity, but A875 NGFr and truncated NGFr in COS cells did not display positive cooperativity. The anti-human NGFr monoclonal antibody NGFR5 was characterized and found not to compete with NGF for binding to NGFr. Fabs were prepared from NGFR5 and 192, an anti-rat NGFR monoclonal antibody that was previously shown not to compete with NGF for binding. Fluorescein-labeled Fabs were used to measure the distribution and lateral diffusion of the NGFr. NGFr expressed on COS and A875 cells are diffusely distributed, but NGFr on the surface of PC12 cells appeared, for some cells, to be patched. In A875 cells, 51% of the NGFr was free to diffuse with diffusion coefficient (D) approximately 7 X 10(-10) cm2/s. In COS cells, 43% diffused with D approximately 5 X 10(-10) cm2/s. There was no significant difference in diffusibility between the full-length NGFr and the truncated NGFr. We compared NGFr diffusion on PC12 cells in suspension or adherent to collagen-coated coverslips. For suspension cells, we obtained 32% recovery with D approximately 2.5 X 10(-9) cm2/s. On adherent cells, we obtained 17% recovery with 6 X 10(-9) cm2/s. Binding of NGF enhanced lateral diffusion of NGFr in A875 cells and in PC12 cells in suspension but did not alter lateral diffusion of NGFr in COS cells or in adherent PC12 cells. NGF had no effect on the diffusing fraction or the distribution of NGFR for any cell line.

scholarly journals Selective destruction of nerve growth factor receptor-bearing cells in vitro using a hybrid toxin composed of ricin A chain and a monoclonal antibody against the nerve growth factor receptor.

1985 ◽  
Vol 101 (3) ◽  
pp. 1107-1114 ◽  
Author(s):  
P S DiStefano ◽  
J B Schweitzer ◽  
M Taniuchi ◽  
E M Johnson
Keyword(s):  
Protein Synthesis ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Growth Factor Receptor ◽  
Ricin A Chain ◽  
Nerve Growth ◽  
Ngf Receptor ◽  
A Chain ◽  
Ricin A

A hybrid toxin composed of ricin A chain and a monoclonal antibody directed against the rat nerve growth factor (NGF) receptor (192-IgG) was prepared using the heterobifunctional cross-linking agent N-succinimidyl-3-(2-pyridyldithio)-propionate and purified by affinity chromatography. Characterization studies showed that the hybrid, 192-s-s-A, displaced bound 125I-labeled 192-IgG from rat superior cervical ganglion (SCG) membranes with an IC50 3-5 times lower than that of unconjugated 192-IgG. When incubated with cultured rat SCG neurons, 192-s-s-A inhibited protein synthesis in a concentration-dependent fashion. The effect of 192-s-s-A on these neurons was reversed by coincubation with an excess of 192-IgG. The IC50 of 192-s-s-A on protein synthesis in rat SCG neurons was 4 nM. Intact ricin and ricin A chain inhibited protein synthesis in these neurons with IC50 values of 5 pM and 500 nM, respectively. The 192-s-s-A hybrid had no effect on mouse SCG neurons or a human melanoma cell line known to have NGF receptors. This is consistent with the finding that 192-IgG recognizes only the rat NGF receptor. Also, 192-s-s-A did not inhibit protein synthesis in primary cultures of rat skeletal muscle or Vero cells, which do not have cell surface receptors for NGF. 192-s-s-A was able to inhibit protein synthesis in PC12 cells but the potency was 10-100 times less in these cells compared to rat SCG neurons. Ricin and A chain were also 10-100 times less potent in PC12 cells than neurons. Rat SCG neurons exposed to 192-s-s-A lost their refractile appearance under phase-contrast optics, showed granular degeneration of neurites, and died. Thus the decreased protein synthesis caused by the hybrid toxin correlated with the morphological destruction of the neurons. 192-s-s-A represents a potentially powerful tool by which to selectively destroy NGF receptor-bearing cells in vitro. The hybrid toxin may prove useful as an in vivo toxin.

scholarly journals Circulating nerve growth factor receptor positive cells are associated with severity and prognosis of pulmonary arterial hypertension

2021 ◽  
Vol 11 (1) ◽  
pp. 204589402199052
Author(s):  
Chiaki Goten ◽  
Soichiro Usui ◽  
Shin-ichiro Takashima ◽  
Oto Inoue ◽  
Hirofumi Okada ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Pulmonary Arterial Hypertension ◽  
Arterial Hypertension ◽  
Growth Factor Receptor ◽  
Nerve Growth Factor Receptor ◽  
Clinical Severity ◽  
Hemodynamic Parameters ◽  
Nerve Growth ◽  
Pulmonary Arterial

Pulmonary arterial hypertension (PAH) remains a disease with a poor prognosis, so early detection and treatment are very important. Sensitive and non-invasive markers for PAH are urgently required. This study was performed to identify sensitive markers of the clinical severity and prognosis of PAH. Patients diagnosed with PAH (n = 30) and control participants (n = 15) were enrolled in this observational study. Major EPC and MSC markers (including CD34, CD133, VEGFR2, CD90, PDGFRα, and NGFR) in peripheral blood mononuclear cells (PBMNCs) were assessed by flow cytometry. Associations of these markers with hemodynamic parameters (e.g. mean pulmonary arterial pressure, pulmonary vascular resistance, and cardiac index) were assessed. Patients with PAH were followed up for 12 months to assess the incidence of major adverse events, defined as death or lung transplantation. Levels of circulating EPC and MSC markers in PBMNCs were higher in patients with PAH than in control participants. Among the studied markers, nerve growth factor receptor (NGFR) was significantly positively correlated with hemodynamic parameters. During the 12-month follow-up period, major-event-free survival was significantly higher in patients with PAH who had relatively low frequencies of NGFR positive cells than patients who had higher frequencies. These results suggested that the presence of circulating NGFR positive cells among PBMNCs may be a novel biomarker for the severity and prognosis of PAH.

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