scholarly journals Use of a promoter-trap retrovirus to identify and isolate genes involved in differentiation of a myeloid progenitor cell line in vitro

Blood ◽  
1996 ◽  
Vol 87 (5) ◽  
pp. 1771-1779 ◽  
Author(s):  
JI Jonsson ◽  
Q Wu ◽  
K Nilsson ◽  
RA Phillips
Keyword(s):  
Dna Sequences ◽  
Progenitor Cell ◽  
High Efficiency ◽  
Sequence Similarity ◽  
Lacz Gene ◽  
Developmentally Regulated ◽  
Myeloid Progenitor ◽  
Myeloid Progenitor Cell ◽  
Promoter Trap

Abstract Studies of gene regulation during early hematopoiesis and of the regulatory network that controls differentiation and lineage commitment are hampered by difficulties in isolating and growing stem cells and early progenitor cells. These difficulties preclude the application of standard molecular genetic approaches to these problems. As an alternative approach we have introduced a lacZ-containing promoter-trap retrovirus into hematopoietic cells. We used the interleukin-3- dependent mouse myeloid progenitor cell 32D as a model to identify transcriptionally active genes. The frequency of integrations that led to transcription of the lacZ gene was estimated to be 0.5% of all integrations, of which 14% were downregulated on differentiation of 32D cells towards neutrophils. Thus, one in every 1,000 to 2,000 integrations identified a developmentally regulated gene. Cellular DNA sequences upstream of proviral integrations were isolated by inverse polymerase chain reaction. Five were further characterized and we confirmed by RNA expression analysis that they were downregulated on differentiation. Sequence analysis revealed identification of novel genes with sequence similarity to known genes. Considering the high efficiency of retroviral infection, our study shows the feasibility of using promoter-trap vectors to identity and isolate developmentally regulated genes from early hematopoietic progenitors.

scholarly journals Use of a promoter-trap retrovirus to identify and isolate genes involved in differentiation of a myeloid progenitor cell line in vitro

Blood ◽  
1996 ◽  
Vol 87 (5) ◽  
pp. 1771-1779 ◽  
Author(s):  
JI Jonsson ◽  
Q Wu ◽  
K Nilsson ◽  
RA Phillips
Keyword(s):  
Dna Sequences ◽  
Progenitor Cell ◽  
High Efficiency ◽  
Sequence Similarity ◽  
Lacz Gene ◽  
Developmentally Regulated ◽  
Myeloid Progenitor ◽  
Myeloid Progenitor Cell ◽  
Promoter Trap

Studies of gene regulation during early hematopoiesis and of the regulatory network that controls differentiation and lineage commitment are hampered by difficulties in isolating and growing stem cells and early progenitor cells. These difficulties preclude the application of standard molecular genetic approaches to these problems. As an alternative approach we have introduced a lacZ-containing promoter-trap retrovirus into hematopoietic cells. We used the interleukin-3- dependent mouse myeloid progenitor cell 32D as a model to identify transcriptionally active genes. The frequency of integrations that led to transcription of the lacZ gene was estimated to be 0.5% of all integrations, of which 14% were downregulated on differentiation of 32D cells towards neutrophils. Thus, one in every 1,000 to 2,000 integrations identified a developmentally regulated gene. Cellular DNA sequences upstream of proviral integrations were isolated by inverse polymerase chain reaction. Five were further characterized and we confirmed by RNA expression analysis that they were downregulated on differentiation. Sequence analysis revealed identification of novel genes with sequence similarity to known genes. Considering the high efficiency of retroviral infection, our study shows the feasibility of using promoter-trap vectors to identity and isolate developmentally regulated genes from early hematopoietic progenitors.

IL-31 Receptor and IL-31 in the Positive Regulation of Myeloid Progenitor Cell Proliferation/Survival.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1347-1347
Author(s):  
Hal E. Broxmeyer ◽  
Nico Ghilardi ◽  
Giao Hangoc ◽  
Scott Cooper ◽  
Wen Tao ◽  
...  
Keyword(s):  
Colony Formation ◽  
Progenitor Cell ◽  
S Phase ◽  
Oncostatin M ◽  
Littermate Control ◽  
Gm Csf ◽  
Myeloid Progenitor ◽  
Myeloid Progenitor Cell

Abstract Interleukin (IL)-31 receptor (R), also called gp130-like monocyte-receptor (GLM-R; Ghilardi et al. J. Biol. Chem.277:16831, 2002) is related to gp130 (~25% homology), and G-CSF-R (~24%). Its signaling activates STAT3 and STAT5. IL-31 is a four helix bundle cytokine preferentially produced by T helper 2 cells. Nothing is known of the possible hematopoietic effects of IL-31R and IL-31. However, since: IL-31 signals through a receptor composed of IL-31RA and Oncostatin M R (Dillon et al. Nature Immunol. 5:752,2004), Oncostatin M is implicated in homeostasis of myeloid progenitor cells (Broxmeyer et al. Immunity.16:815, 2002), and STAT3 and 5 are implicated by a number of groups in cytokine regulation of hematopoiesis, we hypothesized that the IL-31/IL-31R axis would be involved in regulation of hematopoiesis. We first compared myeloid progenitor cell (MPC: CFU-GM, BFU-E, and CFU-GEMM) numbers and cycling status in marrow and spleen of IL-31R −/− vs. littermate control mice (+/+) using a combination of Epo, SCF and PWMSCM to stimulate in vitro the cells taken from these mice. IL-31R −/− mice had significantly decreased numbers of MPC per femur (~51%) and spleen (~36–45%) with significantly decreased MPC cycling status in marrow (% MPC in S-phase: 0–3% in IL-31R −/− vs. 41–53% in +/+ mice). MPC in spleen of IL-31 −/− and +/+ were both in a slow or non cycling state (0–3% in S-phase). In contrast to CFU-GM from +/+ mice, CFU-GM from IL-31R −/− mice demonstrated little or no synergistic response to combined stimulation in vitro with GM-CSF or IL-3 with either SCF or Flt3-ligand (FL). This translated to decreased absolute numbers per femur of GM-CSF+SCF-, GM-CSF+FL-, IL-3+SCF-, and IL-3+FL- responsive CFU-GM in IL-31R −/− mice. However, there were no significant differences between GM-CSF- or IL-3- responsive CFU-GM per femur between IL-31R −/− vs. +/+ mice suggesting effects on immature subsets of MPC. Recombinant IL-31 was assessed for effects in vitro. IL-31, at concentrations up to 100ng/ml, did not stimulate colony formation by marrow MPC, nor did it enhance or suppress colony formation stimulated by GM-CSF, Epo, Epo+SCF, or Epo+SCF+GM-CSF. However, IL-31 did enhance survival of MPC subjected to delayed growth factor addition in a manner similar to, but not as potent as, that of SDF-1/CXCL12. IL-31 manifested no chemotaxis activity for +/+ MPC, and IL-31R −/− and +/+ MPC were equally responsive to the chemotactic effects of SDF-1/CXCL12. These results suggest that the IL-31R in vivo acts to positively regulate numbers and cycling of immature subsets of MPC in the marrow, and that IL-31 has survival enhancing effects on MPC in vitro.

scholarly journals Normal myeloid progenitor cell subset-associated gene signatures for acute myeloid leukaemia subtyping with prognostic impact

PLoS ONE ◽  
2020 ◽  
Vol 15 (4) ◽  
pp. e0229593 ◽  
Author(s):  
Anna A. Schönherz ◽  
Julie Støve Bødker ◽  
Alexander Schmitz ◽  
Rasmus Froberg Brøndum ◽  
Lasse Hjort Jakobsen ◽  
...  
Keyword(s):  
Acute Myeloid Leukaemia ◽  
Myeloid Leukaemia ◽  
Progenitor Cell ◽  
Cell Subset ◽  
Prognostic Impact ◽  
Gene Signatures ◽  
Myeloid Progenitor ◽  
Myeloid Progenitor Cell ◽  
Acute Myeloid

scholarly journals Acute leukemia associated with the t(4;11) chromosome rearrangement: ultrastructural and immunologic characteristics

Blood ◽  
1982 ◽  
Vol 60 (6) ◽  
pp. 1321-1331 ◽  
Author(s):  
JL Parkin ◽  
DC Arthur ◽  
CS Abramson ◽  
RW McKenna ◽  
JH Kersey ◽  
...  
Keyword(s):  
Acute Leukemia ◽  
Clinical Features ◽  
Progenitor Cell ◽  
Chromosome Rearrangement ◽  
Poor Response ◽  
Response To Therapy ◽  
Leukemic Cells ◽  
Myeloid Progenitor ◽  
Hla Dr ◽  
Myeloid Progenitor Cell

Abstract The acute leukemia associated with the t(4;11) chromosome rearrangement is characterized by relatively consistent clinical features: occurrence primarily in young individuals, hyperleukocytosis, and poor response to therapy. This study describes the morphological, ultrastructural, and immunologic characteristics of the leukemic cells from ten patients with this type of leukemia. The morphological features of the leukemic blasts vary from lymphoid-appearing to monocytic. Ultrastructurally and cytochemically, some of the lymphoid-appearing blasts possess features of myeloid origin. The immunologic phenotype is characteristically E- SIg- CALLA- BA-1- BA-2+ HLA-DR+ and TdT+. These findings suggest that the t(4;11)-associated acute leukemia represents a proliferation of an early myeloid progenitor cell.

Malaria toxins from P. chabaudi chabaudi AS and P. berghei ANKA cause dyserythropoiesis in C57BL/6 mice

Parasitology ◽  
1997 ◽  
Vol 115 (5) ◽  
pp. 467-474 ◽  
Author(s):  
W. RUDIN ◽  
V. QUESNIAUX ◽  
N. FAVRE ◽  
G. BORDMANN
Keyword(s):  
Severe Malaria ◽  
Plasmodium Berghei ◽  
Intraperitoneal Injection ◽  
Proliferative Activity ◽  
Progenitor Cell ◽  
Myeloid Progenitor ◽  
Flow Cytometric ◽  
Myeloid Progenitor Cell ◽  
Colony Assays ◽  
Daily Intraperitoneal Injection

The lack of correlation between parasitaemia and anaemia in severe malaria indicates that factors in addition to schizont rupture or erythrophagocytosis contribute to anaemia. We asked whether malaria toxin (MT) from Plasmodium berghei or P. chabaudi might impair erythropoiesis. Daily intraperitoneal injection of MT into C57BL/6 mice induced a transient reduction of RBC values by 25–30% after about 2 weeks, followed by increased haematopoiesis in the spleen as compared to mice receiving uninfected RBC preparations. There was a 3 (P. berghei) to 8-fold (P. chabaudi) increase of total proliferative activity in the spleen. Flow cytometric analyses showed that this was accompanied by some differentiation of TER-119 positive erythroid cells and of Gr-1 positive myeloid cells. Erythroid and myeloid progenitor cell-derived colony assays confirmed these results and revealed an increase in the number of CFU-E ([les ]200-fold), BFU-E ([les ]10-fold) and CFU-GM ([les ]20-fold) in the spleen of MT treated mice, as compared to controls.

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