Mammalian cell production and purification of progenipoietin, a dual-agonist chimaeric haematopoietic growth factor

2003 ◽  
Vol 37 (1) ◽  
pp. 31 ◽  
Author(s):  
David C. Wood ◽  
Karl J. Mathis ◽  
William D. Joy ◽  
John C. Minnerly ◽  
Lyle E. Pegg ◽  
...  
Keyword(s):  
Growth Factor ◽  
Mammalian Cell ◽  
Cell Production ◽  
Haematopoietic Growth Factor ◽  
Dual Agonist

scholarly journals Processing and secretion of nerve growth factor: expression in mammalian cells with a vaccinia virus vector.

1988 ◽  
Vol 8 (6) ◽  
pp. 2456-2464 ◽  
Author(s):  
R H Edwards ◽  
M J Selby ◽  
W C Mobley ◽  
S L Weinrich ◽  
D E Hruby ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cyclic Amp ◽  
Vaccinia Virus ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Signal Peptide ◽  
Virus Vector ◽  
Mammalian Cell Lines ◽  
Vaccinia Virus Vector

To study posttranslational mechanisms for the control of nerve growth factor (NGF), we used a recombinant vaccinia virus vector to independently express the two major NGF transcripts in a variety of mammalian cell lines. The two major transcripts contain NGF (12.5 kilodaltons [kDa]) at the C-terminus and differ by alternative splicing of an N-terminal exon, so that the large precursor (34 kDa) had 67 amino acids upstream of an internal signal peptide and the smaller precursor (27 kDa) had this signal peptide at its N-terminus. In L929 cells, expression of either NGF transcript with the vaccinia virus vector gave rise to an apparently identical intracellular 35-kDa glycosylated precursor formed by cleavage of the primary gene product after the signal peptide. These cells also secreted biologically active NGF. To determine whether NGF processing is restricted by cell type, we infected a variety of mammalian cell lines with both recombinant viruses; all accumulated the same 35-kDa precursor and secreted NGF. Thus, many types of cells have the machinery to process and secrete NGF. However, NGF accumulated intracellularly (presumably in secretory granules) in cells with a regulated pathway of secretion (e.g., AtT-20 and HIT cells). In these cells, a membrane-permeable cyclic AMP analog, 8-bromo-cyclic AMP, stimulated NGF secretion. This suggests a mechanism for the regulation of NGF levels in which specific secretagogues, e.g., neurotransmitters, control NGF secretion.

Role for BH3-Only Protein NOXA In Growth-Factor Deprivation and Early Erythropoiesis

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4235-4235
Author(s):  
Christian R. Geest ◽  
Felix M. Wensveen ◽  
Sten F.W.M. Libregts ◽  
Alex M. de Bruin ◽  
Ingrid A.M. Derks ◽  
...  
Keyword(s):  
Growth Factor ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Erythroid Cell ◽  
Erythroid Progenitors ◽  
Erythroid Progenitor ◽  
Apoptosis Pathway ◽  
Cell Production ◽  
Growth Factor Deprivation ◽  
Redundant Role

Abstract Abstract 4235 Red blood cell production is a strictly regulated process and homeostatic maintenance of the erythropoietic system requires equilibrium between the rate of erythroid cell production and red blood cell destruction. Hematopoietic cytokines play a crucial role in regulating expansion, differentiation and survival of erythrocyte progenitors. Shortage of growth factors triggers the mitochondrial apoptosis pathway, which is critically dependent on Bcl-2 family members. However, the contribution of this mechanism in the regulation of erythropoiesis remains ill-defined. This prompted us to screen for candidate genes involved in this process in erythroid progenitors. We found that the expression of Noxa, a pro-apoptotic Bcl-2 family member, is upregulated during erythroid differentiation and following cytokine-withdrawal in erythroid progenitor cells. Knockdown or deletion of Noxa in IL-3 dependent human and murine erythroid progenitor cell lines increased Mcl-1 levels, which correlated with markedly decreased apoptosis following cytokine withdrawal. Importantly, Noxa ablation in mice increased extra-medullary erythropoiesis, resulting in enhanced numbers of early splenic erythroblasts and circulating reticulocytes. Noxa-deficient hematopoietic progenitors were more resistant to apoptosis induced by growth factor deprivation and displayed increased colony-forming potential. In addition, combined loss of Noxa and Bim resulted in enhanced resistance of erythroid progenitors to cytokine withdrawal compared to WT or single Bim knockouts, suggesting a non-redundant role for Noxa and Bim in regulating survival of erythroid progenitors in response to cytokine deprivation. Finally, in a model of acute haemolytic anaemia, deletion of Noxa enhanced subsequent hematocrit recovery. Together, these findings identify a non-redundant role for BH3-only protein Noxa in the regulation of erythroblast survival during early erythropoiesis. Therefore, Noxa may be a novel component to control red blood cell numbers and modulation of this pathway could be envisaged in therapeutic options for treatment of anaemia. Disclosures: No relevant conflicts of interest to declare.

Embryonic expression of a haematopoietic growth factor encoded by the SI locus and the ligand for c-kit

Nature ◽  
1990 ◽  
Vol 347 (6294) ◽  
pp. 667-669 ◽  
Author(s):  
Yasuhisa Matsui ◽  
Kristina M. Zsebo ◽  
Brigid L. M. Hogan
Keyword(s):  
Growth Factor ◽  
Haematopoietic Growth Factor

Analysis of Mammalian Cell Growth Factor Receptor Dynamics

1987 ◽  
Vol 506 (1 Biochemical E) ◽  
pp. 147-162 ◽  
Author(s):  
DOUGLAS A. LAUFFENBURGER ◽  
JENNIFER LINDERMAN ◽  
LYLE BERKOWITZ
Keyword(s):  
Growth Factor ◽  
Cell Growth ◽  
Mammalian Cell ◽  
Growth Factor Receptor ◽  
Receptor Dynamics ◽  
Mammalian Cell Growth ◽  
Cell Growth Factor Receptor

Processing and secretion of nerve growth factor: expression in mammalian cells with a vaccinia virus vector

1988 ◽  
Vol 8 (6) ◽  
pp. 2456-2464
Author(s):  
R H Edwards ◽  
M J Selby ◽  
W C Mobley ◽  
S L Weinrich ◽  
D E Hruby ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cyclic Amp ◽  
Vaccinia Virus ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Signal Peptide ◽  
Virus Vector ◽  
Mammalian Cell Lines ◽  
Vaccinia Virus Vector

To study posttranslational mechanisms for the control of nerve growth factor (NGF), we used a recombinant vaccinia virus vector to independently express the two major NGF transcripts in a variety of mammalian cell lines. The two major transcripts contain NGF (12.5 kilodaltons [kDa]) at the C-terminus and differ by alternative splicing of an N-terminal exon, so that the large precursor (34 kDa) had 67 amino acids upstream of an internal signal peptide and the smaller precursor (27 kDa) had this signal peptide at its N-terminus. In L929 cells, expression of either NGF transcript with the vaccinia virus vector gave rise to an apparently identical intracellular 35-kDa glycosylated precursor formed by cleavage of the primary gene product after the signal peptide. These cells also secreted biologically active NGF. To determine whether NGF processing is restricted by cell type, we infected a variety of mammalian cell lines with both recombinant viruses; all accumulated the same 35-kDa precursor and secreted NGF. Thus, many types of cells have the machinery to process and secrete NGF. However, NGF accumulated intracellularly (presumably in secretory granules) in cells with a regulated pathway of secretion (e.g., AtT-20 and HIT cells). In these cells, a membrane-permeable cyclic AMP analog, 8-bromo-cyclic AMP, stimulated NGF secretion. This suggests a mechanism for the regulation of NGF levels in which specific secretagogues, e.g., neurotransmitters, control NGF secretion.

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