Regulation of Erythroid Differentiation: Characterization of Rauscher and Friend Virus-Transformed Proerythroid Cells

1979 ◽  
Vol 160 (1) ◽  
pp. 24-27 ◽  
Author(s):  
R. M. Miao ◽  
A. H. Fieldsteel
Keyword(s):  
Erythroid Differentiation ◽  
Friend Virus

scholarly journals Isolation of erythropoietin-sensitive cells from Friend virus-infected marrow cultures: characteristics of the erythropoietin response

Blood ◽  
1983 ◽  
Vol 61 (4) ◽  
pp. 751-758 ◽  
Author(s):  
M Bondurant ◽  
M Koury ◽  
SB Krantz ◽  
T Blevins ◽  
DT Duncan
Keyword(s):  
Terminal Differentiation ◽  
Erythroid Differentiation ◽  
Precursor Cells ◽  
Erythroid Cells ◽  
Friend Virus ◽  
Isolated Cells ◽  
Two Dimensional Gel Electrophoresis ◽  
Erythroid Precursor ◽  
Short Time

Abstract Murine erythroid precursor cells, stimulated to proliferate in vitro in the absence of added erythropoietin (EP) by the anemia strain of Friend virus (FVA), will subsequently respond to EP by complete erythrocyte differentiation. If not exposed to EP, the erythroid cells divide for about 120 hr in culture, and they maintain the potential for full differentiation in response to EP added at any time during the period from 72 to 120 hr. Between 96 and 120 hr of culture without added EP, the EP-sensitive erythroid precursor cells that have formed discrete erythroid bursts can be isolated in relatively large numbers from such cultures by plucking with a Pasteur pipette. The addition of EP initiates the final stages of erythroid differentiation, including heme synthesis in 70%-80% of these isolated cells. With respect to homogeneity of the precursor cells, quantity of EP-responsive cells obtainable, and uniformity of EP responsiveness, this system is uniquely favorable for biochemical studies of the late differentiation effects of EP. The overall changes in gene expression accompanying EP- induced terminal differentiation were examined by two-dimensional gel electrophoresis of proteins labeled for a short time with radioactive amino acids. Several new proteins are synthesized in these erythroid cells during terminal differentiation, but the number is a very small percentage of the total number of proteins being made. Thus, in this system, the effect of EP is to initiate expression of a small group of genes, including those for globins, spectrin, and other proteins involved in the final stages of erythroid differentiation.

Characterization of HOX Gene Expression During Myelopoiesis: Role of HOX A5 in Lineage Commitment and Maturation

Blood ◽  
1999 ◽  
Vol 93 (10) ◽  
pp. 3391-3400 ◽  
Author(s):  
John F. Fuller ◽  
Jeanne McAdara ◽  
Yifah Yaron ◽  
Mark Sakaguchi ◽  
John K. Fraser ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Bone Marrow Cells ◽  
Homeobox Genes ◽  
Erythroid Differentiation ◽  
Erythroid Progenitors ◽  
Effector Cells ◽  
Gm Csf ◽  
Important Regulator ◽  
Macrophage Colony

During the process of normal hematopoiesis, proliferation is tightly linked to maturation. The molecular mechanisms that lead to production of mature effector cells with a variety of phenotypes and functions from a single multipotent progenitor are only beginning to be elucidated. It is important to determine how these maturation events are regulated at the molecular level, because this will provide significant insights into the process of normal hematopoiesis as well as leukemogenesis. Transcription factors containing the highly conserved homeobox motif show considerable promise as potential regulators of hematopoietic maturation events. In this study, we focused on identification and characterization of homeobox genes of the HOX family that are important in regulating normal human myeloid differentiation induced by the hematopoietic growth factor, granulocyte-macrophage colony-stimulating factor (GM-CSF). We have identified three homeobox genes, HOX A5, HOX B6, and HOX B7, which are expressed during early myelopoiesis. Treating bone marrow cells with antisense oligodeoxynucleotides to HOX A5 resulted in inhibition of granulocytic/monocytic hematopoiesis and increased the generation of erythroid progenitors. Also, overexpression of HOX A5 inhibited erythroid differentiation of the K562 cell line. Based on these observations, we propose that HOX A5 functions as an important regulator of hematopoietic lineage determination and maturation.

Establishment and characterization of a new erythroblastic leukemia cell line, EEB: Phosphatidylglucoside-mediated erythroid differentiation and apoptosis

2006 ◽  
Vol 30 (7) ◽  
pp. 829-839 ◽  
Author(s):  
Chizuru Kawano-Yamamoto ◽  
Kazuo Muroi ◽  
Yasuko Nagatsuka ◽  
Masato Higuchi ◽  
Satoru Kikuchi ◽  
...  
Keyword(s):  
Cell Line ◽  
Leukemia Cell ◽  
Erythroid Differentiation ◽  
Leukemia Cell Line

scholarly journals Characterization of a Live-Attenuated Retroviral Vaccine Demonstrates Protection via Immune Mechanisms

1998 ◽  
Vol 72 (8) ◽  
pp. 6554-6558 ◽  
Author(s):  
Ulf Dittmer ◽  
Diane M. Brooks ◽  
Kim J. Hasenkrug
Keyword(s):  
Genetic Resistance ◽  
Helper Virus ◽  
Vaccine Virus ◽  
Friend Virus ◽  
Live Attenuated Vaccine ◽  
Immune Mechanisms ◽  
Viral Interference ◽  
Adult Mice ◽  
Virus Complex

ABSTRACT Live-attenuated retroviruses have been shown to be effective retroviral vaccines, but currently little is known regarding the mechanisms of protection. In the present studies, we used Friend virus as a model to analyze characteristics of a live-attenuated vaccine in protection against virus-induced disease. Highly susceptible mice were immunized with nonpathogenic Friend murine leukemia helper virus (F-MuLV), which replicates poorly in adult mice. Further attenuation of the vaccine virus was achieved by crossing the Fv-1 genetic resistance barrier. The minimum dose of vaccine virus required to protect 100% of the mice against challenge with pathogenic Friend virus complex was determined to be 103 focus-forming units of attenuated virus. Live vaccine virus was necessary for induction of immunity, since inactivated F-MuLV did not induce protection. To determine whether immune cells mediated protection, spleen cells from vaccinated donor mice were adoptively transferred into syngeneic recipients. The results indicated that immune mechanisms rather than viral interference mediated protection.

Characterization of Multiple Isoforms of Protein 4.1R Expressed During Erythroid Terminal Differentiation

Blood ◽  
1998 ◽  
Vol 92 (11) ◽  
pp. 4404-4414 ◽  
Author(s):  
P. Gascard ◽  
G. Lee ◽  
L. Coulombel ◽  
I. Auffray ◽  
M. Lum ◽  
...  
Keyword(s):  
Primary Structure ◽  
Splice Variants ◽  
Cell Lineage ◽  
Erythroid Differentiation ◽  
Membrane Properties ◽  
Exon 2 ◽  
Erythroid Terminal Differentiation ◽  
And Function ◽  
Protein 4.1R

In erythrocytes, 80-kD protein 4.1R regulates critical membrane properties of deformability and mechanical strength. However, previously obtained data suggest that multiple isoforms of protein 4.1, generated by alternative pre-mRNA splicing, are expressed during erythroid differentiation. Erythroid precursors use two splice acceptor sites at the 5′ end of exon 2, thereby generating two populations of 4.1 RNA: one that includes an upstream AUG-1 in exon 2′ and encodes high molecular weight isoforms, and another that skips AUG-1 in exon 2′ and encodes 4.1 by initiation at a downstream AUG-2 in exon 4. To begin an analysis of the complex picture of protein 4.1R expression and function during erythropoiesis, we determined the number and primary structure of 4.1R isoforms expressed in erythroblasts. We used reverse-transcription polymerase chain reaction to amplify and clone full-length coding domains from the population of 4.1R cDNA containing AUG-1 and the population excluding AUG-1. We observed an impressive repertoire of 4.1R isoforms that included 7 major and 11 minor splice variants, thus providing the first definitive characterization of 4.1R primary structures in a single-cell lineage. 4.1R isoforms, transfected into COS-7 cells, distributed to the nucleus, cytoplasm, plasma membrane, and apparent centrosome. We confirmed previous studies showing that inclusion of exon 16 was essential for efficient nuclear localization. Unexpectedly, immunochemical analysis of COS-7 cells transfected with an isoform lacking both AUG-1 and AUG-2 documented that a previously unidentified downstream translation initiation codon located in exon 8 can regulate expression of 4.1R. We speculate that the repertoire of primary structure of 4.1R dictates its distinct binding partners and functions during erythropoiesis.

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