Evaluation of veterinary autogenous vaccines safety by an MTT in-vitro cytotoxicity assay
DOI:
https://doi.org/10.12834/VetIt.1778.9390.2Keywords:
Veterinary autogenous vaccines, ATT-MTT assays, Alternative methodsAbstract
In Italy, veterinary autogenous vaccines manufacturing is regulated by the legislative decree of the Ministry of Health, March 17th, 1994, n. 287. The production is performed by the network of the ‘Istituti Zooprofilattici Sperimentali’ (IZSs), public health institutes scattered all over the Italian territory. The aim of this research was to evaluate the feasibility of an in vitro method to test the abnormal toxicity of autogenous bacterial vaccines as an alternative to animal models routinely employed. For this purpose, the Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise (IZSAM) in partnership with the Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna (IZSLER), evaluated the toxicity of 49 batches of autogenous bacterial vaccines, previously shown to be safe in guinea pigs and mice, on animal model, by means of the methyl tetrazolium (MTT) assay. All vaccines showed cytotoxic effects when tested 1:2 diluted and undiluted; overall, all vaccines lost toxicity at 1:128 dilution. As expected, these findings suggest a different susceptibility of this assay compared to the laboratory animal model. On the other hand, these results do not clarify which components of the vaccines are responsible for the cytotoxic effect. Overall, more experiments are warranted in order to standardize the MTT assay which could be coupled with the trials in laboratory animals.References
Badole G P, Madhukar M. M., Meshram G.K., Bahadure R. N., Tawani S., Tawani G. & Badole S.G. 2013. A comparative evaluation of cytotoxicity of root canal sealers: an in vitro study. 2013. Restorative Dentistry Endodontics, 38 (4), 204-209.
Ciuchini F., Fischetti R. & Piccininno G. 1986. Manuale per il controllo di qualità di sieri, vaccini e prodotti diagnostici di origine batterica, per uso veterinario. Roma, Istituto Superiore di Sanità (Rapporti ISTISAN 86/13).
Combrisson H. 2014. The Directive 2010/63/UE: the explicit and the implicit. Bull Acad Vet France, 167 (2), 137-142.
Di Francesco C.E., Leone A., Lombari V., Luciani M. & Paladini C. 2005. Cell-mediated response in cattle experimentally infected with bluetongue virus serotype 2. Vet Ital, 41 (1), 22-33.
Evans B.R. & Leighton F.A. 2014. A history of One Health. Rev Sci Tech, 33 (2), 413-420.Fabre I. 2009. Alternatives to animal testing. Bull Acad Natl Med, 193 (8), 1783-1791.
Faller C. & Bracher M. 2002. Reconstructed skin kits: reproducibility of cutaneous irritancy testing. Skin Pharmacol Appl Skin Physiol, 15 (1), 74-91.
Fotakis G. & Timbrell J.A. 2006. In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicology Letters,160 (2), 171-177.
Kaplan B., Kahn L.H. & Monath T.P. 2009. One Health - One Medicine: linking human, animal and environmental health The brewing storm. Vet Ital, 45 (1), 9-18.
Kumar S., Bharti V.K., Kumar P. & Pandey R.P. 2018. Proposed deletion of abnormal toxicity test and 3rs initiatives for other safety test of biologicals. Source Journal of Pharmaceutical Sciences, 1 (1), 1-16.
Lin W., Zhang G., Cao P., Zhang D., Zheng Y., Wu R., Qin L., Wang G. & Wen T. 2015. Cytotoxicity and its test methodology for a bioabsorbable nitrided iron stent. J Biomed Materials Res, 103 (4), 764-776.
Malkoc S., Corekci B., Esra Ulker H., Yalçin M. & Sengu A. 2010. Cytotoxic effects of orthodontic composites. Angle Orthodontist, 80 (4), 759-763. Monath T.P. 2013. Vaccines against diseases transmitted from animals to humans: a one health paradigm. Vaccine, 31, 5321-5338.
Monography n. 20609/2008. Abnormal toxicity. European Pharmacopoeia Ed.9.0 Vol. 1 pag. 194.
Monography n. 50209/2013. Evaluation of safety of each batch of veterinary vaccines and immunosera. European Pharmacopoeia Ed.9.0 Vol. 1 pag. 625.
Monography n. 20418/2008. Free formaldheyde. European Pharmacopoeia Ed.9.0 Vol. 1 pag. 137.
Mosmann T. 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods, 65 (1-2), 55-63.
Nordin M., Wieslander A., Martinson E. & Kjellstrand P. 1991. Effects of exposure period of acetylsalicylic acid, paracetamol and isopropanol on L929 cytotoxicity. Toxicol in Vitro, 5 (5), 449-450.
Organisation for Economic Co-operation and Development (OECD). 2015. Test No. 439: In vitro skin irritation: reconstructed human epidermis test method, OECD Guidelines for the testing of chemicals, Section 4. OECD Publishing, Paris, 1-21. https://doi.org/10.1787/9789264242845-en.
Ohtake T., Maeda Y., Hayashi T., Yamanaka H., Nakai M. & Takeyoshi M. 2018. Applicability of an integrated testing strategy consisting of in silico, in chemico and in vitro assays for evaluating the skin sensitization potencies of isocyanates. Toxicology, 15, 393-399.
Patnaik R. & Padhy R.N. 2018. Comparative study on toxicity of methylmercury chloride and methylmercury hydroxide to the human neuroblastoma cell line SH-SY5Y. Environ Sci Pollut Res Int, May 11. doi: 10.1007/s11356-018-2164.
Qi F., Yan Q., Zheng Z., Liu J., Chen Y. & Zhang G. 2018. Geraniol and geranyl acetate induce potent anticancer effects in colon cancer Colo-205 cells by inducing apoptosis, DNA damage and cell cycle arrest. J BUON, 23 (2), 346-352.
Russell W.M.S. & Burch R.L. 1959 (as reprinted 1992). The principles of humane experimental technique. Wheathampstead (UK): Universities Federation for Animal Welfare.
Schwanig M., Nagel M., Duchow K. & Krämer B. 1997. Elimination of abnormal toxicity test for sera and certain vaccines in the European Pharmacopoeia. Vaccine, 15 (10), 1047-1048.
Standard operating procedure: “Autogenous vaccines production and control” 2001. Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise. IZS TEB5.1.2 SOP008.
Thibault E. 2004. Current situation and use of autogenous vaccines in France: the views of the manufacturer and the user. Dev Biol, 117, 73-81.
Thonemann B., Schmalz G., Hiller K.A & Schweikl H. 2002. Responses of L929 mouse fibroblasts, primary and immortalized bovine dental papilla-derived cell lines to dental resin components. Dental Materials, 18 (4), 318-323.
World Organisation for Animal Health (OIE). 2018. Principles of veterinary vaccine production. Chapter 1.1.8. In Manual of diagnostic tests and vaccines for terrestrial animals. 7th Ed. OIE, Paris