Title: Carbamylated-BSA is immunologically active and autoantibodies in sera of diabetic nephropathy patients show strong binding towards it

Authors: Dr Rubana Kauser, Dr Md Arif Iquabal, Dr Md Faizur Rahman, Dr Sangita Choudhary, Tanweer Md Iqbal

 DOI: https://dx.doi.org/10.18535/jmscr/v10i10.16

Abstract

Isocyanates (ICN) have drawn considerable attention in the recent past as they can react with macromolecules (polypeptides, nucleic acids etc.) and produce toxic insults. ICN react with side chain nitrogen of amino acid residues of polypeptides. Pathophysiological implications resulting from occupational and accidental exposures of ICN are yet too elusive. It may be produced in high concentrations in conditions of chronic renal failure and chronic inflammatory diseases (diabetes mellitus, rheumatoid arthritis etc.). In this study, bovine serum albumin (BSA) has been carbamylated and characterized by UV and AGE. Antibodies against carbamylated-BSA were raised in animals and characterized by direct binding and inhibition ELISA. Presence of anti-carbamylated-BSA autoantibodies in sera of diabetic nephropathy patients was elucidated by ELISA. Carbamylated-BSA exhibited hyperchromicity at 280 nm compared to native BSA. Carbamylated-BSA was found to be highly immunogenic as compared to native BSA. Experimental induction of antibodies against carbamylated-BSA and presence of autoantibodies against carbamylated-BSA in diabetic nephropathic patients’ sera points towards the role for carbamylated-BSA in diabetic nephropathy.

Keywords: Isocyanates; Carbamylated-BSA; Autoantibodies; Diabetic nephropathy patients.

References

  1. H. Karol, Target organs and systems: Methodologies to assess immune system function, Environ. Health Perspect.106 (1998) 533–540.
  2. Vojdani, M. Ghoneum, N. Brautbar, Immune alteration associated with exposure to toxic chemicals, Toxicol. Ind. Health 8 (1992) 239–254.
  3. D. Shelby, J.W. Allen, W.J. Caspary, S. Haworth, J. Ivett, A. Kligerman, C.A. Luke, J.M. Mason, B. Myhr, R.R. Tice, R. Valencia, E. Zeiger, Results of in vitro and in vivo genetic toxicity tests on methyl isocyanate, Environ. Health Perspect.72 (1987) 183–187.
  4. Tamura, K. Aoki, M.S. Lee, Characterization and genotoxicity of DNA adducts caused by 2-naphthyl isocyanate, Carcinogenesis 11 (1990) 2009–2014.
  5. W. Worthy, Methyl isocyanate: the chemistry of a hazard, Chem. Eng. News 63 (1985) 27–33.
  1. Caspary, B. Myhr, Mutagenicity of methyl isocyanate and its reaction products to cultured mammalian cells, Mutat. Res. 174 (1986) 285–293.
  2. Segal, J.J. Solomon, F.J. Life, Isolation of methylcarbamyl adducts of adenine and cytosine following in vitro reaction of methyl isocyanate with calf thymus DNA, Chem. Biol. Interact. 69 (1989) 359–372.
  3. Tamura, K. Aoki, M.S. Lee, Selective reactivities of isocyanates towards DNA bases and genotoxicity of methyl carbamylation of DNA, Mutat. Res. 283 (1992) 97–106.
  4. P. Baumann, H.A. Seow, K. Shyam, P.G. Penketh, A.C. Sartorelli, The antineoplastic efficacy of the prodrug cloretazine is produced by the synergistic interaction of carbamylating and alkylating products of its activation, Oncol. Res. 15 (2005) 313‒325.
  5. Ali,K. Alam, Evaluation of antibodies against free radical-modified DNA by ELISA, in: D. Armstrong (Ed.), Oxidative Stress Biomarkers and Antioxidant Protocols: Methods Mol. Biol., Humana Press Inc., New Jersey, 2002, pp. 171‒181.
  6. Alam, Moinuddin, S. Jabeen, Immunogenicity of mitochondrial DNA modified by hydroxyl radical, Cell. Immunol. 247 (2007) 12‒17.
  7. T. Marczynski, L. Shapiro, Cell-cycle control of a cloned chromosomal origin of replication from Caulobacter crescentus, Mol. Biol. 226 (1992) 959‒977.
  8. Beyerbach, P.B. Farmer, G. Sabbioni, Biomarkers for isocyanate exposure: Synthesis of isocyanate DNA adducts, Chem. Res. Toxicol. 19 (2006) 1611–1618.
  9. C. Yoon, P. Puigserver, G. Chen, J. Donovan, Z. Wu, Control of hepatic gluconeogenesis through the transcriptional coactivator PGC-1, Nature (London)413 (2001) 131‒138.
  10. G. Deo, S. Gangal, A.N. Bhisey, R. Somasundaram, B. Balsara, B. Gulwani, Immunological, mutagenic & genotoxic investigations in gas exposed population of Bhopal, Indian J. Med. Res. 86 (1987) 63–76.
  11. K. Saxena, K.P. Singh, S.L. Nagle, B.N. Gupta, P.K. Ray, R.K. Srivastav, Effect of exposure to toxic gas on the population of Bhopal: Part IV-Immunological and chromosomal studies, Indian J. Exp. Biol. 26 (1988) 173–176.
  12. K. Goswami, Cytogenetic effects of methyl isocyanate exposure in Bhopal, Adv. Hum. Genet. 74 (1986) 81–84.
  13. N. Chen, L. Porubleva, G. Shearer, M. Svrakic, L.G. Holden, J.L. Dover, M. Johnston, P.R. Chitnis, D.H. Kohl, Associating protein activities with their genes: Rapid identification of a gene encoding a methylglyoxal reductase in the yeast, Yeast 20 (2003) 545‒554.
  14. Hu, B.R. Kim, C. Chen, V. Hebbar, A.N. Kong, The roles of JNK and apoptotic signaling pathways in PEITC-mediated responses in human HT-29 colon adenocarcinoma cells, Carcinogenesis 24 (2003) 1361–1367.
  15. Zhang, L. Tang, V. Gonzalez, Selected isothiocyanates rapidly induce growth inhibition of cancer cells, Mol. Cancer Ther. 2 (2003) 1045–1052.
  16. Tang, Y. Zhang, Mitochondria are the primary target in isothiocyanates induced apoptosis in human bladder cancer cells, Mol. Cancer Ther. 4 (2005) 1250–1259.
  17. J. East, C.N. Abboud, R.F. Borch, Diethyldithiocarbamate induction of cytokine release in human long-term bone marrow cultures, Blood 80 (1992) 1172–1177.
  18. P. Hussain, L.J. Hofseth, C.C. Harris, Radical causes of cancer, Nature Rev. Cancer 3 (2003) 276–285.
  19. K. Hurst, W.C. Barrette, Leukocytic oxygen activation and microbicidal oxidative toxins, Crit. Rev. Biochem. Mol. Biol. 24 (1989) 271–328.
  20. L. Ramos, S. Pou, B.E. Britigan, M.S. Cohen, G.M. Rosen, Spin trapping evidence for myeloperoxidase-dependent hydroxyl radical formation by human neutrophils and monocytes, J. Biol. Chem. 267 (1992) 8307–8312.
  21. J. Steineck, A.U. Khan, M.J. Karnovsky, Extracellular production of singlet oxygen by stimulated macrophages quantified using 9,10-diphenylanthracene and perylene in a polystyrene film, J. Biol. Chem. 267 (1992) 13425–13433.
  22. Nakamura, A. Murakami, Y. Ohto, K. Torikai, T. Tanaka, H. Ohigashi, Suppression of tumor promoter-induced oxidative stress and inflammatory responses in mouse skin by a superoxide generation inhibitor 1′-acetoxychavicol acetate, Cancer Res. 58 (1998) 4832–4839.
  23. S. Ahmad, Moinuddin, K. Dixit, U. Shahab, K. Alam, A. Ali, Genotoxicity and immunogenicity of DNA-advanced glycation end products formed by methylglyoxal and lysine in presence of Cu2+, Biochem. Biophys. Res. Commun. 407 (2011) 568–574.
  24. G.I. Dovbeshko, N.Y. Gridina, E.B. Kruglova, O.P. Pashchu, FTIR spectroscopy studies of nucleic acid damage, Talanta 53 (2000) 233‒246.
  25. J.K. Pijanka, A. Kohler, Y. Yang, P. Dumas, S. Chio-Srichan, M. Manfait, G.D. Sockalingum, J. Sulé-Suso, Spectroscopic signatures of single, isolated cancer cell nuclei using synchrotron infrared microscopy, Analyst 134 (2009) 1176.
  1. Polak, J. Lekki, O. Veselov, Z. Stachura, J. Styczeń, Single proton hit facility at the IFJ PAN in Cracow, Acta Phys. Pol. 109 (2006) 417‒419.
  2. Ugenskiene, J. Lekki, W. Polak, M. Prise, M. Folkard, O. Veselov, Z. Stachura, W.M. Kwiatek, M. Zazula, J. Stachura, Double strand break formation as a response to X-ray and targeted proton-irradiation, Instrum. Methods Phys. Res. 260 (2007) 159‒163.
  3. Alam, A. Ali, R. Ali, The effect of hydroxyl radical on the antigenicity of native DNA, FEBS Lett. 319 (1993) 66–70.
  4. Waris, K. Alam, Immunogenicity of superoxide radical modified-DNA: Studies on induced antibodies and SLE anti-DNA autoantibodies, Life Sci. 75 (2004) 2633–2642.
  5. Ahmad, Moinuddin, S. Habib, U. Shahab, K. Alam, A. Ali, Autoimmune response to AGE modified human DNA: Implications in type 1 diabetes mellitus, J. Clin. Trans. Endocrinol.1 (2014) 66‒72.

Corresponding Author

Dr Sangita Choudhary, M.D.

Department of Biochemistry, Katihar Medical College, Al-Karim University, Katihar, Bihar, 854109, India