Production of a monoclonal antibody directed against the nerve growth factor receptor from sympathetic membranes

1985 ◽  
Vol 27 (2) ◽  
pp. 121-132 ◽  
Author(s):  
Claudia J. Morgan ◽  
Ralph A. Bradshaw
Keyword(s):  
Monoclonal Antibody ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Growth Factor Receptor ◽  
Nerve Growth Factor Receptor ◽  
Nerve Growth

Production of a Monoclonal Antibody Directed against the High-Affinity Nerve Growth Factor Receptor

1999 ◽  
Vol 14 (2) ◽  
pp. 68-72 ◽  
Author(s):  
E. Tagliabue ◽  
C. Ghirelli ◽  
L. Lombardi ◽  
F. Castiglioni ◽  
L. Asnaghi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Neuroblastoma Cell ◽  
Growth Factor Receptor ◽  
Nerve Growth Factor Receptor ◽  
Receptor Internalization ◽  
Growth Modulation ◽  
Nerve Growth ◽  
High Affinity

The high-affinity nerve growth factor receptor corresponds to the tyrosine protein kinase encoded by the proto-oncogene trkA. Different findings suggest that nerve growth factor (NGF) can be operative in the growth modulation of tumor cell lines possessing high-affinity binding sites for this molecule. Using as immunizing material the SKNBE neuroblastoma cell line transfected with proto-trkA we produced a monoclonal antibody (MAb) able to recognize the high-affinity nerve growth factor receptor. The selected MAb, designated MGR12, is directed against an epitope present on the extracellular domain of the receptor since it showed reactivity on living trkA-expressing cells and was able to immunoprecipitate the proto-trkA molecule. The MGR12 MAb is directed against a non-functional epitope since it neither inhibited NGF binding nor induced receptor internalization. This new reagent appears to be an appropriate tool for analyzing the expression of high-affinity nerve growth factor receptor in tumors of different origin and for elucidating its involvement in tumor progression.

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 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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