Radiation and Environmental Biophysics

, Volume 49, Issue 3, pp 405–419 | Cite as

Dose-rate effects of protons on in vivo activation of nuclear factor-kappa B and cytokines in mouse bone marrow cells

  • Kanokporn Noy Rithidech
  • Paiboon Reungpatthanaphong
  • Louise Honikel
  • Adam Rusek
  • Sanford R. Simon
Original Paper


The objective of this study was to determine the kinetics of nuclear factor-kappa B (NF-κB) activation and cytokine expression in bone marrow (BM) cells of exposed mice as a function of the dose rate of protons. The cytokines included in this study are pro-inflammatory [i.e., tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), and IL-6] and anti-inflammatory cytokines (i.e., IL-4 and IL-10). We gave male BALB/cJ mice a whole-body exposure to 0 (sham-controls) or 1.0 Gy of 100 MeV protons, delivered at 5 or 10 mGy min−1, the dose and dose rates found during solar particle events in space. As a reference radiation, groups of mice were exposed to 0 (sham-controls) or 1 Gy of 137Cs γ rays (10 mGy min−1). After irradiation, BM cells were collected at 1.5, 3, 24 h, and 1 month for analyses (five mice per treatment group per harvest time). The results indicated that the in vivo time course of effects induced by a single dose of 1 Gy of 100 MeV protons or 137Cs γ rays, delivered at 10 mGy min−1, was similar. Although statistically significant levels of NF-κB activation and pro-inflammatory cytokines in BM cells of exposed mice when compared to those in the corresponding sham controls (Student’s t-test, p < 0.05 or <0.01) were induced by either dose rate, these levels varied over time for each protein. Further, only a dose rate of 5 mGy min−1 induced significant levels of anti-inflammatory cytokines. The results indicate dose-rate effects of protons.


Dose Rate Bone Marrow Cell Sham Control High Dose Rate Brookhaven National Laboratory 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank Dr. Peter Guida and his team for logistic support, MaryAnn Petry and her BLAF staff for their assistance in animal handling. We also thank Dr. Michael Sivertz for dosimetry support. This work was supported by the National Aeronautics and Space Administration (NASA) Grant #NNX07AP88G.


  1. Albanese J, Martens K, Karkanitsa LV, Dainiak N (2007) Multivariate analysis of low-dose radiation-associated changes in cytokine gene expression profiles using microarray technology. Exp Hematol 35:47–54CrossRefGoogle Scholar
  2. Baldwin AS Jr (1996) The NF-kappa B and I kappa B proteins: new discoveries and insights. Annu Rev Immunol 14:649–683CrossRefGoogle Scholar
  3. Barendsen GW (1968) Responses of cultured cells, tumours and normal tissues to radiations of different linear enerygy transfer. Curr Top Radiat Res 4:293–356Google Scholar
  4. Barendsen GW (1994) RBE-LET relationships for different types of lethal radiation damage in mammalian cells: comparison with DNA dsb and an interpretation of differences in radiosensitivity. Int J Radiat Biol 66:433–436CrossRefGoogle Scholar
  5. Barton BE (1997) IL-6: insights into novel biological activities. Clin Immunol and Immunopathol 85:16–20CrossRefGoogle Scholar
  6. Basu S, Rosenzweig KR, Youmell M, Price BD (1998) The DNA-dependent protein kinase participates in the activation of NF[kappa]B following DNA damage. Biochem Biophys Res Commun 247:79–83CrossRefGoogle Scholar
  7. Baumstark-Khan C, Hellweg CE, Arenz A, Meie MA (2005) Cellular monitoring of the nuclear factor κB pathway for assessment of space environmental radiation. Radiat Res 164:527–530CrossRefGoogle Scholar
  8. BEIR (2005) Health risks from exposure to low levels of ionizing radiation: BEIR VII Phase 2 vol BEIR VII. The National AcademiesGoogle Scholar
  9. Boreham DR, Dolling JA, Maves SR, Siwarungsun N, Mitchel RE (2000) Dose-rate effects for apoptosis and micronucleus formation in gamma-irradiated human lymphocytes. Radiat Res 153:579–586CrossRefGoogle Scholar
  10. Brach MA, Hass R, Sherman ML, Gunji H, Weichselbaum R, Kufe D (1991) Ionizing radiation induces expression and binding activity of the nuclear factor kappa B. J Clin Invest 88:691–695CrossRefGoogle Scholar
  11. Cassatella MA, Meda L, Gasperini S, Calzetti F, Bonora S (1994) Interleukin 10 (IL-10) upregulates IL-1 receptor antagonist production from lipopolysaccharide-stimulated human polymorphonuclear leukocytes by delaying mRNA degradation. J Exp Med 179:1695–1699CrossRefGoogle Scholar
  12. Catley MC, Chivers JE, Cambridge LM, Holden N, Slater DM, Staples KJ, Bergmann MW, Loser P, Barnes PJ, Newton R (2003) IL-1[beta]-dependent activation of NF-[kappa]B mediates PGE2 release via the expression of cyclooxygenase-2 and microsomal prostaglandin E synthase. FEBS Lett 547:75–79CrossRefGoogle Scholar
  13. Chang CM, Elliott TB, Dobson ME, Jackson WE, Ledney GD (2000) Ionizing radiation and bacterial challenge alter splenic cytokine gene expression. J Radiat Res 41:259–277CrossRefGoogle Scholar
  14. Chang PY, Bjornstad KA, Rosen CJ, McNamara MP, Mancini R, Goldstein LE, Chylack LT, Blakely EA (2005) Effects of iron ions, protons and X rays on human lens cell differentiation. Radiat Res 164:531–539CrossRefGoogle Scholar
  15. Cleghorn TF, Saganti PB, Zeitlin CJ, Cucinotta FA (2004) Solar particle events observed at Mars: dosimetry measurements and model calculations. Adv Space Res 33:2215–2218CrossRefADSGoogle Scholar
  16. Cornforth MN, Bailey SM, Goodwin EH (2002) Dose responses for chromosome aberrations produced in noncycling primary human fibroblasts by alpha particles, and by gamma rays delivered at sublimiting low dose rates. Radiat Res 158:43–53CrossRefGoogle Scholar
  17. Cucinotta FA (1999) Issues in risk assessment from solar particle events. Radiat Meas 30:261–268CrossRefGoogle Scholar
  18. Durante M, Cucinotta FA (2008) Heavy ion carcinogenesis and human space exploration. Nat Rev Cancer 8:465–472CrossRefGoogle Scholar
  19. Elmore E, Lao XY, Ko M, Rightnar S, Nelson G, Redpath J (2005) Neoplastic transformation in vitro induced by low doses of 232 MeV protons. Int J Radiat Biol 81:291–297CrossRefGoogle Scholar
  20. Elmore E, Lao XY, Kapadia R, Redpath JL (2006) The effect of dose rate on radiation-induced neoplastic transformation in vitro by low doses of low-LET radiation. Radiat Res 166:832–838CrossRefGoogle Scholar
  21. Fan M, Ahmed KM, Coleman MC, Spitz DR, Li JJ (2007) Nuclear factor-{kappa}B and manganese superoxide dismutase mediate adaptive radioresistance in low-dose irradiated mouse skin epithelial cells. Cancer Res 67:3220–3228CrossRefGoogle Scholar
  22. Geard CR, Chen CY (1990) Micronuclei and clonogenicity following low- and high-dose-rate gamma irradiation of normal human fibroblasts. Radiat Res 124:S56–S61CrossRefGoogle Scholar
  23. Ghosh S, May MJ, Kopp EB (1998) NF-kappa B and Rel proteins: evolutionarily conserved mediators of immune responses. Annu Rev Immunol 16:225–260CrossRefGoogle Scholar
  24. Green LM, Murray DK, Bant AM, Kazarians G, Moyers MF, Nelson GA, Tran DT (2001) Response of thyroid follicular cells to gamma irradiation compared to proton irradiation. I. Initial characterization of DNA damage, micronucleus formation, apoptosis, cell survival, and cell cycle phase redistribution. Radiat Res 155:32–42CrossRefGoogle Scholar
  25. Gueulette J, Böhm L, Slabbert JP, De Coster BM, Rutherfoord GS, Ruifrok A, Octave-Prignot M, Binns PJ, Schreuder AN, Symons JE, Scalliet P, Jones DTL (2000) Proton relative biological effectiveness (RBE) for survival in mice after thoracic irradiation with fractionated doses. Int J Radiat Oncol Biol Phys 47:1051–1058Google Scholar
  26. Hada M, Sutherland BM (2006) Spectrum of complex DNA damages depends on the incident radiation. Radiat Res 165:223–230CrossRefGoogle Scholar
  27. Hellweg CE, Arenz A, Meier MM, Baumstark-Khan C (2005) Cellular monitoring systems for the assessment of space environmental factors. Adv Space Res 36:1673–1679CrossRefADSGoogle Scholar
  28. Hosoi Y, Miyachi H, Matsumoto Y, Enomoto A, Nakagawa K, Suzuki N, Ono T (2001) Induction of interleukin-1beta and interleukin-6 mRNA by low doses of ionizing radiation in macrophages. Int J Cancer 96:270–276CrossRefGoogle Scholar
  29. Hu S, Kim M-HY, McClellan GE, Cucinotta FA (2009) Modeling the acute health effects of astronauts from exposure to large solar particle events. Health Phys 96:465–476CrossRefGoogle Scholar
  30. Kato TA, Nagasawa H, Weil MM, Genik PC, Little JB, Bedford JS (2006) gamma-H2AX foci after low-dose-rate irradiation reveal atm haploinsufficiency in mice. Radiat Res 166:47–54CrossRefGoogle Scholar
  31. Kim M-HY, Hayat MJ, Feiveson AH, Cucinotta FA (2009) Prediction of frequency and exposure level of solar particle events. Health Phys 97:68–81. doi: 10.1097/01.HP.0000346799.65001.9c CrossRefGoogle Scholar
  32. Kumar PR, Mohankumar MN, Hamza VZ, Jeevanram RK (2006) Dose-rate effect on the induction of HPRT mutants in human G0 lymphocytes exposed in vitro to gamma radiation. Radiat Res 165:43–50CrossRefGoogle Scholar
  33. Lancel S, Bachschmid MM, Kirber MT, Weinberg EO (2008) Abstract 3378: IL-33 translocates to the nucleus and has NF-kB transcriptional repressor function following treatment with IL-1beta in human endothelial cells. Circulation 118:S_415-aGoogle Scholar
  34. Linard C, Ropenga A, Vozenin-Brotons MC, Chapel A, Mathe D (2003) Abdominal irradiation increases inflammatory cytokine expression and activates NF-{kappa}B in rat ileal muscularis layer. Am J Physiol Gastrointest Liver Physiol 285:G556–G565Google Scholar
  35. Linard C, Marquette C, Mathieu J, Pennequin A, Clarencon D, Mathe D (2004) Acute induction of inflammatory cytokine expression after [gamma]-irradiation in the rat: effect of an NF-[kappa]B inhibitor. Int J Radiat Oncol Biol Phys 58:427–434Google Scholar
  36. Little MP, Muirhead CR (1997) Curvilinearity in the dose-response curve for cancer in Japanese atomic bomb survivors. Environ Health Perspect 105(Suppl 6):1505–1509CrossRefGoogle Scholar
  37. Liu Z-g, Hsu H, Goeddel DV, Karin M (1996) Dissection of TNF receptor 1 effector functions: JNK activation is not linked to apoptosis while NF-[kappa]B activation prevents cell death. Cell 87:565–576CrossRefGoogle Scholar
  38. Liu XD, Ma SM, Liu SZ (2003) Effects of 0.075 Gy x-ray irradiation on the expression of IL-10 and IL-12 in mice. Phys Med Biol 48:2041–2049CrossRefGoogle Scholar
  39. Lorimore SA, Coates PJ, Wright EG (2003) Radiation-induced genomic instability and bystander effects: inter-related nontargeted effects of exposure to ionizing radiation. Oncogene 22:7058–7069CrossRefGoogle Scholar
  40. Loucas BD, Eberle R, Bailey SM, Cornforth MN (2004) Influence of dose rate on the induction of simple and complex chromosome exchanges by gamma rays. Radiat Res 162:339–349CrossRefGoogle Scholar
  41. Mohan N, Meltz ML (1994) Induction of nuclear factor kappa B after low-dose ionizing radiation involves a reactive oxygen intermediate signaling pathway. Radiat Res 140:97–104CrossRefGoogle Scholar
  42. Moore SR, Marsden S, Macdonald D, Mitchell S, Folkard M, Michael B, Goodhead DT, Prise KM, Kadhim MA (2005) Genomic instability in human lymphocytes irradiated with individual charged particles: involvement of tumor necrosis factor alpha in irradiated cells but not bystander cells. Radiat Res 163:183–190CrossRefGoogle Scholar
  43. Natarajan M, Aravinda N, Meltz M, Herman T (2002) Post-translational modification of I-kappa B alpha activates NF-?B in human monocytes exposed to 56Fe ions. Radiat Environ Biophys 41:139–144Google Scholar
  44. Opal SM, DePalo VA (2000) Anti-inflammatory cytokines. Chest 117:1162–1172CrossRefGoogle Scholar
  45. Parsons JL, Townsend LW (2000) Interplanetary crew dose rates for the August 1972 solar particle event. Radiat Res 153:729–733CrossRefGoogle Scholar
  46. Pikarsky E, Porat RM, Stein I, Abramovitch R, Amit S, Kasem S, Gutkovich-Pyest E, Urieli-Shoval S, Galun E, Ben-Neriah Y (2004) NF-[kappa]B functions as a tumour promoter in inflammation-associated cancer. Nature 431:461–466CrossRefADSGoogle Scholar
  47. Prasad AVM, Mohan N, Meltz ML (1994) Activation of nuclear factor kappa B in human lymphoblastoid cells by low-dose ionizing radiation. Radiat Res 138:367–372CrossRefGoogle Scholar
  48. Preston DL, Mattsson A, Holmberg E, Shore R, Hildreth NG, Boice JD Jr (2002) Radiation effects on breast cancer risk: a pooled analysis of eight cohorts. Radiat Res 158:220–235CrossRefGoogle Scholar
  49. Rho H-S, Kim S-H, Lee C-E (2005) Mechanism of NF-kB activation induced by gamma-irradiation in B-lymphoma cells: role of ras. J Toxicol Environ Health 68:2019–2031CrossRefGoogle Scholar
  50. Rithidech K, Tungjai M, Arbab E, Simon SR (2005) Activation of NF-kappa B in bone marrow cells of BALB/cJ mice following exposure in vivo to low doses of 137Cs gamma rays. Radiat Env Biophys 44:139–143CrossRefGoogle Scholar
  51. Ross HJ, Canada AL, Antoniono RJ, Redpath JL (1997) High and low dose rate irradiation have opposing effects on cytokine gene expression in human glioblastoma cell lines. Eur J Cancer 33:144–152CrossRefGoogle Scholar
  52. Rube C, Uthe D, Wilfert F, Ludwig D, Yang K, Konig J, Palm J, Schuck A, Willich N, Remberger K, Rube C (2005) The bronchiolar epithelium as a prominent source of pro-inflammatory cytokines after lung irradiation. Int J Radiat Oncol Biol Phys 61:1482–1492Google Scholar
  53. Schreiber S, Nikolaus S, Hampe J (1998) Activation of nuclear factor kappaB in inflammatory bowel disease. Gut 42:477–484CrossRefGoogle Scholar
  54. Shea MA, Smart DF (1994) Recent and historical solar proton eventsGoogle Scholar
  55. Simpson JA (1983) Elemental and isotopic composition of the galactic cosmic rays. Annu Rev Nucl Part Sci 33:323CrossRefADSGoogle Scholar
  56. Smart DF, Shea M (1989) Solar proton events during the past three solar cycles. J Spacecr Rockets 26:403–415CrossRefADSGoogle Scholar
  57. Smart DF, Shea MA (2002) A review of solar proton events during the 22nd solar cycle. Adv Space Res 30:1033–1044CrossRefADSGoogle Scholar
  58. Todorov VT, Volkl S, Muller M, Bohla A, Klar J, Kunz-Schughart LA, Hehlgans T, Kurtz A (2004) Tumor Necrosis Factor-alpha activates NF kappa B to inhibit renin transcription by targeting cAMP-responsive element. J Biol Chem 279:1458–1467CrossRefGoogle Scholar
  59. Verma IM, Stevenson JK, Schwarz EM, Van Antwerp D, Miyamoto S (1995) Rel/NF-kappa B/I kappa B family: intimate tales of association and dissociation. Genes Dev 9:2723–2735CrossRefGoogle Scholar
  60. Wang Y, Meng A, Lang H, Brown SA, Konopa JL, Kindy MS, Schmiedt RA, Thompson JS, Zhou D (2004) Activation of nuclear factor kappaB in vivo selectively protects the murine small intestine against ionizing radiation-induced damage. Cancer Res 64:6240–6246CrossRefGoogle Scholar
  61. Weichselbaum RR, Hallahan D, Fuks Z, Kufe D (1994) Radiation induction of immediate early genes: effectors of the radiation-stress response. Int J Radiat Oncol Biol Phys 30:229–234Google Scholar
  62. Wilson JW, Anderson BM, Cucinotta FA, Ware J, Zeitlin CJ (2006, July) Spacesuit radiation shield design methods. Paper presented at the International Conference on Environmental Systems, NorfolkGoogle Scholar
  63. Zapp EN, Ramsey CR, Townsend LW, Badhwar GD (1999) Solar particle event dose and dose-rate distributions: parameterization of dose-time profiles, with subsequent dose-rate analysis. Radiat Meas 30:393–400CrossRefGoogle Scholar
  64. Zhou D, Brown S, Yu T, Chen G, Barve S, Kang BC (1999a) A high dose of ionizing radiation induces tissue-specific activation of nuclear factor-kappaB in vivo. Rad Res 151:703–709CrossRefGoogle Scholar
  65. Zhou D, Brown SA, Yu T, Chen G, Barve S, Kang BC, Thompson JS (1999b) A high dose of ionizing radiation induces tissue-specific activation of nuclear factor-kappaB in vivo. Radiat Res 151:703–709CrossRefGoogle Scholar
  66. Zhou D, Yu T, Chen G, Brown SA, Yu Z, Mattson MP, Thompson JS (2001) Effects of NF-kappaB1 (p50) targeted gene disruption on ionizing radiation-induced NF-kappaB activation and TNFalpha, IL-1alpha, > IL-1beta and IL-6 mRNA expression in vivo. Int J Radiat Biol 77:763–772CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Kanokporn Noy Rithidech
    • 1
  • Paiboon Reungpatthanaphong
    • 1
    • 2
  • Louise Honikel
    • 1
  • Adam Rusek
    • 3
  • Sanford R. Simon
    • 1
  1. 1.Pathology DepartmentStony Brook UniversityStony BrookUSA
  2. 2.Radio-Isotope Department, Faculty of SciencesKasetsart UniversityBangkokThailand
  3. 3.Accelerator Department, NASA Research LaboratoryBrookhaven National LaboratoryUptonUSA

Personalised recommendations