DETERMINATION OF RICIN IN ENVIRONMENTAL SAMPLES USING BIOASSAY

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Abstract

A bioassay of ricin toxicity in environmental samples using real-time cell index monitoring is proposed. The halfmaximal inhibitory concentration (IC50) of ricin was estimated at 6,7 ng/ml for human hepatoma HepaRG cells proliferation. The antibodies to A- and B-subunits in HepaRG cell media lead to cytoprotective and antiapoptotic effects against the cytotoxicity of ricin. The antibodies neutralised activation of JNK kinase (phosphorylated at Thr183/Tyr185) and prevented accumulation of the active forms of caspase 8 (hydrolysed to Asp384) and caspase 9 (hydrolysed to Asp315) induced by ricin in HepaRG cells. The tested antibodies also prevented a decrease in the intracellular levels of the active forms of Akt 1 kinase (phosphorylated at Ser473) and transcription factor p53 (phosphorylated at Ser46) caused by ricin. The bioassay with antibodies can be considered as a specific method for identifying the toxin in environmental samples.

About the authors

N. Yu. Rogovskaya

Research Institute of Hygiene, Occupational Pathology and Human Ecology, Federal Medical Biological Agency

Author for correspondence.
Email: niigpech@rihophe.ru

Rogovskaya Nadezhda Yur′evna

188663, Leningrad region

Russian Federation

A. Yu. Gorbunov

Research Institute of Hygiene, Occupational Pathology and Human Ecology, Federal Medical Biological Agency

Email: niigpech@rihophe.ru

Gorbunov Aleksandr Yur′evich

188663, Leningrad region

Russian Federation

Ya. A. Dubrovskii

Research Institute of Hygiene, Occupational Pathology and Human Ecology, Federal Medical Biological Agency

Email: niigpech@rihophe.ru

Dubrovskii Yaroslav Aleksandrovich

188663, Leningrad region

Russian Federation

N. S. Khlebnikova

Research Institute of Hygiene, Occupational Pathology and Human Ecology, Federal Medical Biological Agency

Email: niigpech@rihophe.ru

Khlebnikova Natalia Semenovna

188663, Leningrad region

Russian Federation

V. N. Babakov

Research Institute of Hygiene, Occupational Pathology and Human Ecology, Federal Medical Biological Agency

Email: babakov@rihophe.ru

Babakov Vladimir Nikolaevich

188663, Leningrad region

Russian Federation

References

  1. Kozlov J.V., Sudarkina O.J., Kurmanova A.G. Ribosome-inactivating lectins of plants. Molecular Biology. 2006; 40(4): 635-646 (in Russian).
  2. Bolognesi A, Bortolotti M, Maiello S, Battelli MG, Polito L. Ribosome-Inactivating Proteins from Plants: A Historical Overview. Molecules. 2016; 21(12). pii: E1627.
  3. Bozza W.P., Tolleson W.H., Rivera Rosado L.A., Zhang B. Ricin detection: tracking active toxin. Biotechnol Adv. 2015; 33(1):117-123.
  4. Sinegubova E.O., Dubrovina I.A., Myasnikov V.A. Complex approach to the estimation of cytotoxicity of ribosomeinactivating proteins on the cell model in vitro. Medicine of extreme situations. 2019; 21(S1): 53-62 (in Russian).
  5. Endo Y, Tsurugi K. RNA N-glycosidase activity of ricin A-chain.Mechanism of action of the toxic lectin ricin on eukaryotic ribosomes. J Biol Chem 1987; 262:8128– 30.
  6. Dubrovsky Ya.A., Podolskaya E.P. Peptide toxins identification by MALDI-MS method. Nauchnoe Priborostroenie (Scientific Instumentation) 2010; 20(4): 21-35 (in Russian).
  7. Braun A.V., Tikhomirov L.A., Grechukhin A.P., Rybal’chenko I.V., Taranchenko V.F. Detection of ricin in plant extracts and soil using liquid chromatography–highresolution mass spectrometry. J. Anal. Chem. 2018; 73(8):786-795 (in Russian).
  8. Babakov V.N., Rogovskaya N.Yu., Kurdyukov I.D., Beltyukov P.P., Dulov S.A., Radilov A.S. Effect of arylhydrocarbon receptor agonists and lipopolysaccharide on benzo(a)pyrene genotoxicity markers. Toksikologicheskii vestnik (Toxicological Review). 2019; (3):19-25 (in Russian).
  9. Dubrovskii Ya.A., Gorbunov A.Yu., Rogovskaya N.Yu., Murashko E.A., Beltyukov P.P., Babakov V.N. Proteomic approach of biotoxin detection in environmental samples. In: Proc. Med. Biol. Aspects of Chem. Safety. St. Petersburg. 2018: 25-26 (in Russian).
  10. Griffiths G.D., Leek M.D., Gee D.J. The toxic plant proteins ricin and abrin induce apoptotic changes in mammalian lymphoid tissues and intestine. J Pathol. 1987; 151(3):221-9.
  11. Saxena N., Yadav P., Kumar O. The Fas/ Fas ligand apoptotic pathway is involved in abrin-induced apoptosis. Toxicol Sci. 2013;135(1):103-18.
  12. Fodstad O., Olsnes S., Pihl A. Toxicity, distribution and elimination of the cancer ostatic lectins abrin and ricin after parenteral injection into mice. Br J Cancer. 1976;34(4):418-25.
  13. Ramsden C.S., Drayson M.T., Bell E.B. The toxicity, distribution and excretion of ricin holotoxin in rats. Toxicology. 1989;55(1-2):161-71.
  14. Authier F., Djavaheri-Mergny M., Lorin S., Frénoy J.P., Desbuquois B. Fate and action of ricin in rat liver in vivo: translocation of endocytosed ricin into cytosol and induction of intrinsic apoptosis by ricin B-chain. Cell Microbiol. 2016;18(12):1800-1814.

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Copyright (c) 2020 Rogovskaya N.Y., Gorbunov A.Y., Dubrovskii Y.A., Khlebnikova N.S., Babakov V.N.


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