Interleukin 3 protects murine bone marrow cells from apoptosis induced by DNA damaging agents

Chester Beatty Laboratories, Institute of Cancer Research, London, United Kingdom.
Journal of Experimental Medicine (Impact Factor: 12.52). 11/1992; 176(4):1043-51.
Source: PubMed


Murine bone marrow-derived cells, dependent on interleukin 3 (IL-3) for their growth in culture, undergo programmed cell, or apoptosis, upon cytokine withdrawal. Here it is reported that a variety of DNA damaging agents cause a more rapid onset of apoptosis in a factor-dependent cell line, BAF3, deprived of IL-3. In contrast, when cultured in the presence of IL-3, or other growth promoting factors, BAF3 cells are highly resistant to X-irradiation and the cytotoxic drugs etoposide and cisplatin. Overexpression of the bcl2 gene product also protects BAF3 cells from DNA damage. The presence of IL-3 is not required during the initial events of DNA damage or its repair. In the absence of IL-3, cells still complete the repair of DNA breaks within 15 min, and continue to cycle for 5 h. At this time, IL-3 is necessary to prevent the accelerated onset of DNA cleavage from a G2 arrest point.

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    • "Previous studies confirmed that IL-3 can protect bone marrow cells against radiation-induced apoptosis and regulate the expression of certain oncogenes such as c-myc. In addition, IL-3 protects bone marrow cells against DNA damaging agents [15]. In this study, M-NFS-60 and BM-MNCs cells were treated with either SVPII alone or in combination with IL-3. "
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    ABSTRACT: The previous investigation demonstrated the radioprotective efficacy of peptides isolated from the venom of Buthus Martti Karsch. In this study, the effect of isolated scorpion venom peptide II (SVPII) on irradiated M-NFS-60 cells and mouse bone marrow mononuclear cells (BM-MNCs) was observed. The AlamarBlue cell viability assay, a colony-forming unit (CFU) assay, flow cytometry (FCM), immunofluorescence, and Western blotting were used to evaluate cell proliferation, cell cycle progression, and the expression of the IL-3 receptor (IL-3R) protein in non-irradiated and irradiated cells. Proliferation of irradiated M-NFS-60 cells was significantly accelerated by SPVII, and this effect was further enhanced by co-application of IL-3. Similarly, SPVII increased the number of BM-MNC CFUs and this proliferative effect was greater in the presence of SVPII plus IL-3. In addition, SPVII significantly altered cell cycle progression; SVPII enhanced the fraction of unirradiated M-NFS-60 cells in S phase and the fraction of irradiated M-NFS-60 cells arrested in G2/M. The expression of IL-3R protein by unirradiated M-NFS-60 cells was enhanced significantly by SVPII, and SVPII-induced IL-3R overexpression was 10-fold greater in irradiated M-NFS-60 cells. These results indicated the hematopoietic growth factor (HGF)-like effects of SVPII on irradiated cells, possibly mediated by upregulation of IL-3R.
    Cell and Bioscience 07/2013; 3(1):28. DOI:10.1186/2045-3701-3-28 · 3.63 Impact Factor
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    • "The first two populations were cultured in IL-3 containing media prior to factor withdrawal. As reported previously, removal of IL-3 from control Ba/F3 cells results in loss of cell viability that begins within 24 h after factor withdrawal. (28) Removal of IL-3 from stably transfected cells also resulted in cell death with similar kinetics as control cells. CID withdrawal from CID-selected cells reversed proliferation and resulted in cell death. "
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    ABSTRACT: The main methods for producing genetically engineered cells use viral vectors for which safety issues and manufacturing costs remain a concern. In addition, selection of desired cells typically relies on the use of cytotoxic drugs with long culture times. Here, we introduce an efficient non-viral approach combining the Sleeping Beauty (SB) Transposon System with selective proliferation of engineered cells by chemically induced dimerization (CID) of growth factor receptors. Minicircles carrying a SB transposon cassette containing a reporter transgene and a gene for the F36VFGFR1 fusion protein were delivered to the hematopoietic cell line Ba/F3. Stably-transduced Ba/F3 cell populations with >98% purity were obtained within 1 week using this positive selection strategy. Copy number analysis by quantitative PCR (qPCR) revealed that CID-selected cells contain on average higher copy numbers of transgenes than flow cytometry-selected cells, demonstrating selective advantage for cells with multiple transposon insertions. A diverse population of cells is present both before and after culture in CID media, although site-specific qPCR of transposon junctions show that population diversity is significantly reduced after selection due to preferential expansion of clones with multiple integration events. This non-viral, positive selection approach is an attractive alternative for producing engineered cells.
    Nucleic Acids Research 03/2012; 40(11):e85. DOI:10.1093/nar/gks213 · 9.11 Impact Factor
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    • "de la combinaison de cytokines 4F a été élaboré selon deux critères : facteurs de survie et d'expansion cellulaire (Borge et al. 1997; Brandt et al. 1994; Collins et al. 1992; Gratwohl et al. 1998; Keller et al. 1995; Kinoshita et al. 1995; Limanni et al. 1995; Leary et al. 1992; Neta 1997; Neta et al. 1993; Osada et al. 1999; Shah et al. 1996; Veiby et al. 1996). Le SCF et le FL ont été retenus comme facteurs de survie, l'IL-3 comme facteur d'expansion multilignage et la thrombopoïétine comme facteur mégacaryocytaire. "
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    ABSTRACT: Bone marrow aplasia observed following ionizing radiation exposure (Total Body Irradiation; gamma dose range: 2–10 Gy) is a result, in particular, of the radiation-induced (RI) apoptosis in hematopoietic stem and progenitor cells (HSPC). We have previously shown in a baboon model of mobilized peripheral blood CD34+ cell irradiation in vitro that RI apoptosis in HSPC was an early event, mostly occurring within the first 24 hours, which involves the CD95 Fas pathway. Apoptosis may be significantly reduced with a combination of 4 cytokines (4F): Stem Cell Factor (SCF), FLT-3 Ligand (FL), thrombopoietin (TPO), and interleukin-3 (IL-3), each at 50 ng·mL–1 (15% survival versus <3% untreated cells, 24 h post-irradiation at 2.5 Gy). In this study we show that addition of TNF-alpha(800 IU/ml) induces an increase in 4F efficacy in terms of cell survival 24 h after incubation (26% survival after 24 h irradiation exposure at 2.5 Gy) and amplification (k) of CD34+ cells after 6 days in a serum free culture medium (SFM) (kCD34+ = 4.3 and 6.3 respectively for 4F and successive 4F + TNF-alpha/ 4F treatments). In addition, the 4F combination allows culture on pre-established allogenic irradiated stromal cells in vitro at 4 Gy (kCD34+ = 4.5). Overall this study suggests (i) the potential therapeutic interest for an early administration of anti-apoptotic cytokines with or without hematopoiesis inhibitors (emergency cytokine therapy) and (ii) the feasibility in the accidentally irradiated individual, of autologous cell therapy based on ex vivo expansion in order to perform autograft of residual HSPC collected after the accident.Key words: apoptosis, cytokine, hematopoiesis, irradiation, bone marrow aplasia.[Journal translation]
    Canadian Journal of Physiology and Pharmacology 02/2011; 80(7):700-709. DOI:10.1139/y02-071 · 1.77 Impact Factor
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