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Structure alteration and immunological properties of 60Co-gamma-rays irradiated bothropstoxin-I

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Abstract

In this work, the authors investigated the immunological behavior of bothropstoxin-I (BTHX-1), before and after irradiation process, and also the influence of scavengers substances on protein alterations induced by free radical production. Structural modifications were investigated by SDS-PAGE in reducing or non-reducing conditions. In vitro cytotoxicity assay was performed to test average toxic activities of BTHX-I. BALB/c Isogenic mice were immunized with irradiated or non-irradiated (native) forms of BTHX-I and antibody titers and isotypes were determined by ELISA method. Expression of murine cytokines was analyzed by using expression data obtained by quantitative real-time PCR (qPCR) assays. The results indicate that irradiation of proteins leads to significant structural modifications, and also changes the cytokines profile during immunization process, regarding a suitable approach to new immunogenic production.

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References

  1. Grosh DS, Hoopywood LE (1979) Biological effects of radiation, 2nd edn. Academic Press, New York

    Google Scholar 

  2. Boyer AL, Butler EB, DiPetrillo TA, Engler MJ, Fraass B, Grant W III, Ling CC, Low DA, Mackie TR, Mohan R, Purdy JA, Roach M, Rosenman JG, Verhey LJ, Wong JW, Cumberlin RL, Stone H, Palta R (2001) Intensity-modulated radiotherapy: current status and issues of interest. Int J Radiat Oncol Biol Phys 51(4):880–914

    Google Scholar 

  3. Wales A, Kusel JR (1992) Biochemistry of irradiated parasite vaccines: suggested models for their mode of action. Parasitol Today 8(11):358–363

    Article  CAS  Google Scholar 

  4. Zhou X, Ji WJ, Zhu Y, He B, Li H, Huang TG, Li YM (2007) Enhancement of endogenous defenses against ROS by supra-nutritional level of selenium is more safe and effective than antioxidant supplementation in reducing hypertensive target organ damage. Med Hypotheses 68:952–956

    Article  CAS  Google Scholar 

  5. Araujo HP, Bourguignon SC, Boller MAA, Dias AASO, Lucas EPR, Santos IC, Delgado IF (2008) Potency evaluation of antivenoms in Brazil: the national control laboratory experience between 2000 and 2006. Toxicon 51:502–514

    Article  CAS  Google Scholar 

  6. Nascimento N, Seebart CS, Francis B, Rogero JR, Kaiser II (1996) Toxicon 34(1):123–131

    Article  Google Scholar 

  7. Hawkins AL, Davies MJ (2001) Generation and propagation of radical reactions on proteins. Biochim Biophys Acta 1504:196–219

    Article  CAS  Google Scholar 

  8. Cardi BA, Nascimento N, Andrade HF Jr (1998) Irradiation of Crotalus durissus terrificus crotoxin with 60Co gamma-rays induces its uptake by macrophages through scavenger receptors. Int J Radiat Biol 73(5):557–564

    Article  CAS  Google Scholar 

  9. Delves PJ, Roitt IM (2000) The immune system—first of two parts. Adv Immunol 343:37–49

    CAS  Google Scholar 

  10. Parkin J, Cohen B (2001) An overview of the immune system. Lancet 357:1777–1789

    Article  CAS  Google Scholar 

  11. Grunig G, Banz A, Malefyt RW (2005) Molecular regulation of Th2 immunity by dendritic cells. Pharmacol Ther 106:75–96

    Article  CAS  Google Scholar 

  12. McHeyzer-William M, McHeyzer-William L, Panus J, Pogue-Caley R, Bikah G, Driver D, Eisenbraum M (2003) Helper T-cell-regulated B-cell immunity. Microbes Infect 5:205–212

    Article  CAS  Google Scholar 

  13. Spencer PJ, Aird SD, Boni-Mitake M, Nascimento N, Rogero JR (1998) A single-step purification of bothropstoxin-1. Braz J Med Biol Res 31:1125–1127

    Article  CAS  Google Scholar 

  14. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  15. Harlow E, Lane D (1988) Antibodies. A Laboratory Manual. Cold Spring Harbor Laboratory, NY

  16. 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(4):611–622

    Article  CAS  Google Scholar 

  17. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)). Methods 25(4):402–408

    Article  CAS  Google Scholar 

  18. Kume T, Matsuda T (1995) Changes in structural and antigenic properties of proteins by radiation. Radiat Phys Chem Oxf Engl 1993 46(2):225–231

    Google Scholar 

  19. Moon S, Song KB (2001) Effect of γ-irradiation on the molecular properties of ovalbumin and ovomucoid and protection by ascorbic acid. Food Chem 74:479–483

    Article  CAS  Google Scholar 

  20. Oliveira JCR, Montes OCAH, Duarte MM, Diniz CR, Fortes-Dias CL (2001) Toxicity of South American snake venoms measured by an in vitro cell culture assay. Toxicon 40:321–325

    Article  Google Scholar 

  21. Brewer JM, Richmond J, Alexander J (1994) The demonstration of an essential role for macrophages in the in vivo generation of IgG2a antibodies. Clin Exp Immunol 97:164–171

    Article  CAS  Google Scholar 

  22. Sprent J, Surh CD (2002) T Cell Memory. Annu Rev Immunol 20:551–579

    Article  CAS  Google Scholar 

  23. Pavanelli WR (2007) Immunization with bradikinin induce pain and TH1 standard of immune response. Interbio 1(2):30–37

    Google Scholar 

  24. Hein J, Schellenberg U, Bein G, Hackstein H (2001) Quantification of murine IFN-g mRNA and protein expression: impact of real-time kinetic RT-PCR using SYBR green I dye. Scand J Immunol 54:285–291

    Article  CAS  Google Scholar 

  25. Overbergh L, Giulietti A, Valckx D, Decallonne B, Bouillon R, Mathieu C (2003) The use of real-time reverse transcriptase PCR for the quantification of cytokine gene expression. J Biomol Tech 14(1):33–43

    CAS  Google Scholar 

  26. Hsieh CS, Macatonia SE, O’Garra A, Murphy KM (2006) T cell genetic background determines default T helper phenotype development in vitro. J Exp Med 181:713–721

    Article  Google Scholar 

  27. Murphy KM, Reiner SL (2002) The lineage decisions of helper T cells. Nat Rev Immunol 2:933–944

    Article  CAS  Google Scholar 

  28. Han SK, Song JY, Yun YS, Yi SY (2006) Effect of gamma radiation on cytokine expression and cytokine-receptor mediated STAT activation. Int J Radiat Biol 82(9):686–697

    Article  CAS  Google Scholar 

  29. Anraku M, Kabashima M, Namuraa H, Maruyama T, Otagiri M, Gebicki JM, Furutani N, Tomida H (2008) Antioxidant protection of human serum albumin by chitosan. Int J Biol Macromol 43:159–164

    Article  CAS  Google Scholar 

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Acknowledgments

The authors wish to thank CAPES and CNPq from Brazil; Laboratory of Virology of the IMT-SP by the use of the equipment of Real Time PCR; Center of Technology of Radiation (CTR–IPEN) by the irradiation of samples of BTHX-1.

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Authors and Affiliations

  1. Instituto de Pesquisas Energéticas e Nucleares (IPEN/CNEN-SP), Av. Professor Lineu Prestes 2242, 05508-000, São Paulo, SP, Brazil

    J. A. Baptista, D. P. Vieira, A. J. Galisteo Júnior, O. Z. Higa, M. Casare, C. M. Yonamine, P. Caproni, L. A. Campos, H. F. de Andrade Júnior, P. J. Spencer & N. Nascimento

  2. Instituto de Medicina Tropical de São Paulo, Lab. de Protozoologia, São Paulo, SP, Brazil

    D. P. Vieira, A. J. Galisteo Júnior & H. F. de Andrade Júnior

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  1. J. A. Baptista
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  2. D. P. Vieira
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  3. A. J. Galisteo Júnior
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  4. O. Z. Higa
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  5. M. Casare
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  6. C. M. Yonamine
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  7. P. Caproni
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  8. L. A. Campos
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  9. H. F. de Andrade Júnior
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  10. P. J. Spencer
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  11. N. Nascimento
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Correspondence to J. A. Baptista.

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Baptista, J.A., Vieira, D.P., Galisteo Júnior, A.J. et al. Structure alteration and immunological properties of 60Co-gamma-rays irradiated bothropstoxin-I. J Radioanal Nucl Chem 283, 691–698 (2010). https://doi.org/10.1007/s10967-009-0399-6

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  • DOI: https://doi.org/10.1007/s10967-009-0399-6

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