Abstract
Interferon regulatory factors (IRF-1 and IRF-2) are transcription factors of IRF-family that regulate expression of genes for cytokines, chemokines and growth factors in mammalian cells. IRF-1 and IRF-2 play crucial roles in the differentiation of bone marrow cells for immune response. Bone marrow (BM) is the soft lymphoid organ that contains many types of stem cells and produces different types of cells of the blood and immune system. Genetic alterations and damage of the bone marrow cells can lead to different types of blood and immune system-related diseases including anemia and cancer. We have studied the expression of IRF-1 and IRF-2 during radiation-induced damage and regeneration of bone marrow cells after transplantation of freshly isolated bone marrow cells in the mouse. Cell cycle analysis, colony forming unit-fibroblast (CFU-F) assay and bone marrow histology showed that after radiation-induced damage, the bone marrow transplantation resulted in regeneration of the bone marrow up to 24–35% recovery. Real-time quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) for the mRNA expression showed that IRF-1 and IRF-2 were expressed at higher levels in the bone marrow cells of the irradiated (4.34× fold for IRF-1, and 3.87× fold for IRF-2) compared to control and transplanted (1.13× fold for IRF-1, and 1.12× fold IRF-2) mice and immuno-fluorescence analysis for the protein expression showed that IRF-1 and IRF-2 were expressed at higher levels in the bone marrow cells of the irradiated (2.12× fold for IRF-1 and 1.71× fold for IRF-2) compared to control and transplanted (1.73× fold for IRF-1 and 1.21× fold for IRF-2) mice. Thus, IRF-1 and IRF-2 are sensitive and responsive to radiation-induced damage in the bone marrow cells and may also be involved in the bone marrow regeneration process.
Similar content being viewed by others
References
Tamura T, Yanai H, Savitsky D, Taniguchi T (2008) The IRF family transcription factors in immunity and oncogenesis. Annu Rev Immunol 26:535–584
Taniguchi T, Ogasawara K, Takaoka A, Tanaka N (2001) IRF family of transcription factors as regulators of host defense. Annu Rev Immunol 19:623–655
Upreti M, Rath PC (2005) Expression and DNA binding activity of the recombinant interferon regulatory factor-1 (IRF-1) of mouse. Mol Biol Rep 32:103–116
Romeo G, Fiorucci G, Chiantore MV, Percario ZA, Vannucchi S, Affabris E (2002) IRF-1 as a negative regulator of cell proliferation. J Interferon Cytokine Res 22:39–47
Zhao GN, Jiang DS, Li H (2015) Interferon regulatory factors: at the crossroads of immunity, metabolism, and disease. Biochim Biophys Acta 1852:365–378
Sato T, Onai N, Yoshihara H, Arai F, Suda T, Ohteki T (2009) Interferon regulatory factor-2 protects quiescent hematopoietic stem cells from type I interferon–dependent exhaustion. Nat Med 15:696–700
Perazzio AS, Oliveira JS, Figueiredo VL, Chauffaille ML (2017) Increase of IRF-1 gene expression and impairment of T regulatory cells suppression activity on patients with myelodysplastic syndrome: a longitudinal one-year study. Leuk Res 55:6–17
Travlos GS (2006) Normal structure, function, and histology of the bone marrow. Toxicol Pathol 34:548–565
Zhao E, Xu H, Wang L, Kryczek I, Wu K, Hu Y, Wang G, Zou W (2012) Bone marrow and the control of immunity. Cell Mol Immunol 9:11–19
Yu VW, Scadden DT (2016) Heterogeneity of the bone marrow niche. Curr Opin Hematol 23:331–338
Poncin G, Beaulieu A, Humblet C, Thiry A, Oda K, Boniver J, Defresne MP (2012) Characterization of spontaneous bone marrow recovery after sublethal total body irradiation: importance of the osteoblastic/adipocytic balance. PLoS ONE 7:e30818
Wilke C, Holtan SG, Sharkey L, DeFor T, Arora M, Premakanthan P, Yohe S, Vagge S, Zhou D, Holter Chakrabarty JL, Mahe M, Corvo R, Dusenbery K, Storme G, Weisdorf DJ, Verneris MR, Hui S (2016) Marrow damage and hematopoietic recovery following allogeneic bone marrow transplantation for acute leukemias: effect of radiation dose and conditioning regimen. Radiother Oncol 118:65–71
Cui YZ, Hisha H, Yang GX, Fan TX, Jin T, Li Q, Lian Z, Ikehara S (2002) Optimal protocol for total body irradiation for allogeneic bone marrow transplantation in mice. Bone Marrow Transplant 30:843–849
Duran-Struuck R, Dysko RC (2009) Principles of bone marrow transplantation (BMT): providing optimal veterinary and husbandry care to irradiated mice in BMT studies. J Am Assoc Lab Anim Sci 48:11–22
Liebermann DA, Hoffman B (2009) Good and bad IRF-1: role in tumor suppression versus autoimmune disease. Leuk Res 33:1301–1302
Nascimento FR, Gomes EA, Russo M, Lepique AP (2015) Interferon regulatory factor (IRF)-1 is a master regulator of the cross talk between macrophages and L929 fibrosarcoma cells for nitric oxide dependent tumoricidal activity. PLoS ONE 10:e0117782
Paun A, Pitha PM (2007) The IRF family, revisited. Biochimie 89:744–753
Yanai H, Negishi H, Taniguchi T (2012) The IRF family of transcription factors: inception, impact and implications in oncogenesis. Oncoimmunology 1:1376–1386
Chaudhary JK, Rath PC (2017) A simple method for isolation, propagation, characterization, and differentiation of adult mouse bone marrow-derived multipotent mesenchymal stem cells. J Cell Sci Ther 8:1. https://doi.org/10.4172/2157-7013
Soleimani M, Nadri S (2009) A protocol for isolation and culture of mesenchymal stem cells from mouse bone marrow. Nat Protoc 4:102–106
Zhu H, Guo ZK, Jiang XX, Li H, Wang XY, Yao HY, Zhang Y, Mao N (2010) A protocol for isolation and culture of mesenchymal stem cells from mouse compact bone. Nat Protoc 5:550–560
Roy S, Javed S, Jain SK, Majumdar SS, Mukhopadhyay A (2012) Donor hematopoietic stem cells confer long-term marrow reconstitution by self-renewal divisions exceeding to that of host cells. PLoS ONE 7:e50693
Chaudhary JK, Rath PC (2017) Microgrooved-surface topography enhances cellular division and proliferation of mouse bone marrow-derived mesenchymal stem cells. PLoS ONE 12:e0182128
Riccardi C, Nicoletti I (2006) Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc 1:1458–1461
Baustian C, Hanley S, Ceredig R (2015) Isolation, selection and culture methods to enhance clonogenicity of mouse bone marrow derived mesenchymal stromal cell precursors. Stem Cell Res Ther. https://doi.org/10.1186/s13287-015-0139-5
Quan H, Kim SK, Heo SJ, Koak JY, Lee JH (2014) Optimization of growth inducing factors for colony forming and attachment of bone marrow-derived mesenchymal stem cells regarding bioengineering application. J Adv Prosthodontics 6:379–386
Avwioro G (2011) Histochemical uses of haematoxylin—a review. J Pharma Clin Sci 1:24–34
Cho A, Suzuki S, Hatakeyama J, Haruyama N, Kulkarni AB (2010) A method for rapid demineralization of teeth and bones. Open Dent J 4:223–229
Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55:611–622
Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 3:1101–1108
Cai YJ, Wang WS, Yang Y, Sun LH, Teitelbaum DH, Yang H (2013) Up-regulation of intestinal epithelial cell derived IL-7 expression by keratinocyte growth factor through STAT1/IRF-1, IRF-2 pathway. PLoS ONE 8:e58647
Nadri S, Soleimani M, Hosseni RH, Massumi M, Atashi A, Izadpanah R (2007) An efficient method for isolation of murine bone marrow mesenchymal stem cells. Int J Dev Biol 51:723–729
Ooi YY, Rahmat Z, Jose S, Ramasamy R, Vidyadaran S (2013) Immunophenotype and differentiation capacity of bone marrow-derived mesenchymal stem cells from CBA/Ca, ICR and Balb/c mice. World J Stem Cells 5:34–42
Pleyer L, Valent P, Greil R (2016) Mesenchymal stem and progenitor cells in normal and dysplastic hematopoiesis-masters of survival and clonality? Int J Mol Sci. https://doi.org/10.3390/ijms17071009
Anjos-Afonso F, Bonnet D (2008) Isolation, culture, and differentiation potential of mouse marrow stromal cells. Curr Protoc Stem Cell Biol 7:2B.3.1–2B.3.11. https://doi.org/10.1002/9780470151808.sc02b03s7
Li H, Ghazanfari R, Zacharaki D, Lim HC, Scheding S (2016) Isolation and characterization of primary bone marrow mesenchymal stromal cells. Ann N Y Acad Sci 1370:109–118
Digirolamo CM, Stokes D, Colter D, Phinney DG, Class R, Prockop DJ (1999) Propagation and senescence of human marrow stromal cells in culture: a simple colony-forming assay identifies samples with the greatest potential to propagate and differentiate. Br J Haematol 107:275–281
Pallavicini MG, Redfearn W, Necas E, Brecher G (1997) Rescue from lethal irradiation correlates with transplantation of 10–20 CFU-S-day 12. Blood Cells Mol Dis 23:157–168
Cao X, Wu X, Frassica D, Yu B, Pang L, Xian L, Wan M, Lei W, Armour M, Tryggestad E, Wong J, Wen CY, Lu WW, Frassica FJ (2011) Irradiation induces bone injury by damaging bone marrow microenvironment for stem cells. Proc Natl Acad Sci USA 108:1609–1614
Bruna F, Contador D, Conget P, Erranz B, Sossa CL, Arango-Rodríguez ML (2016) Regenerative potential of mesenchymal stromal cells: age-related changes. Stem cells Int. https://doi.org/10.1155/2016/1461648
Han SK, Song JY, Yun YS, Yi SY (2002) Gamma irradiation-reduced IFN-gamma expression, STAT1 signals, and cell-mediated immunity. J Biochem Mol Biol 35:583–589
Galanty Y, Belotserkovskaya R, Coates J, Polo S, Miller KM, Jackson SP (2009) Mammalian SUMO E3-ligases PIAS1 and PIAS4 promote responses to DNA double-strand breaks. Nature 462:935–939
Pamment J, Ramsay E, Kelleher M, Dornan D, Ball KL (2002) Regulation of the IRF-1 tumour modifier during the response to genotoxic stress involves an ATM-dependent signalling pathway. Oncogene 21:7776–7785
Park SM, Chae M, Kim BK, Seo T, Jang IS, Choi JS, Kim IC, Lee JH, Park J (2010) SUMOylated IRF-1 shows oncogenic potential by mimicking IRF-2. Biochem Biophys Res Commun 391:926–930
Gupta M, Rath PC (2014) Interferon regulatory factor-1 (IRF-1) interacts with regulated in development and DNA damage response 2 (REDD2) in the cytoplasm of mouse bone marrow cells. Int J Biol Macromol 65:41–50
Han KJ, Jiang L, Shu HB (2008) Regulation of IRF2 transcriptional activity by its sumoylation. Biochem Biophys Res Commun 372:772–778
Park J, Kim K, Lee EJ, Seo YJ, Lim SN, Park K, Rho SB, Lee SH, Lee JH (2007) Elevated level of SUMOylated IRF-1 in tumor cells interferes with IRF-1-mediated apoptosis. Proc Natl Acad Sci USA 104:17028–17033
Acknowledgements
Financial supports from the University Grants Commission (UGC)-Research Network Resource Centre (UGC-RNRC) and UGC-DRS as well as the Department of Science and Technology (DST)-FIST (DST-FIST) and DST-PURSE of the Government of India to the School of Life Sciences, J.N.U. and P.C.R., as well as the UGC-JRF & SRF fellowship to N.A. are gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interest related to this manuscript.
Ethical approval
All ethical principles and practice were strictly followed and adhered to with respect to both the experiments on the animals, and the scientific research and publication norms with respect to this manuscript.
Rights and permissions
About this article
Cite this article
Ahamad, N., Rath, P.C. Expression of interferon regulatory factors (IRF-1 and IRF-2) during radiation-induced damage and regeneration of bone marrow by transplantation in mouse. Mol Biol Rep 46, 551–567 (2019). https://doi.org/10.1007/s11033-018-4508-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-018-4508-x