Application of two in vitro methods for the toxicity test of autogenous vaccines


https://doi.org/10.4081/bse.114
Vol. 1 No. 1 (2020)
Published: 14 February 2020
Autogenous vaccines, toxicity tests, in vitro methods, MTT, IL-1β
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Authors

  • Erika Molica Colella
    erika.molicacolella@izsler.it
    Centro di Referenza Nazionale per i Metodi Alternativi, Benessere e Cura degli Animali da Laboratorio, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (IZSLER), Brescia, Italy.
  • Silvia Dotti Centro di Referenza Nazionale per i Metodi Alternativi, Benessere e Cura degli Animali da Laboratorio, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (IZSLER), Brescia, Italy.
  • Riccardo Villa Centro di Referenza Nazionale per i Metodi Alternativi, Benessere e Cura degli Animali da Laboratorio, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (IZSLER), Brescia, Italy.
  • Guerino Lombardi Centro di Referenza Nazionale per la Formazione in Sanità Pubblica Veterinaria, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (IZSLER), Brescia, Italy.
  • Massimo Amadori Laboratorio Benessere Animale, Biochimica Clinica e Immunologia Veterinaria, Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (IZSLER), Brescia, Italy.

According to the 3Rs principle (Replacement, Refinement, Reduction), this study aims to find alternative methods to evaluate the toxicity of autogenous vaccines. Currently in Italy the Istituti Zooprofilattici Sperimentali (II.ZZ.SS.) must perform the in vivo toxicity test for each lot of autogenous vaccine produced as laid down in the Decree of 17 March 1994. This paper describes two in vitro methods for assessing the toxicity of autogenous vaccines. The first is the MTT test based on the metabolic reaction of tetrazolium salt in vital cells. The second method is the test for measurement of IL-1ß production by macrophages, obtained after in vitro differentiation from pig monocytes in peripheral blood mononuclear cells. The two tests were performed on different vaccine antigens dilution: 1:20, 1:100 and 1:500. The results show a positive tendency between the two methods pointing out the potential of these methodologies combined for the replacement of the current in vivo test.


Molica Colella, E., Dotti, S., Villa, R., Lombardi, G., & Amadori, M. (2020). Application of two in vitro methods for the toxicity test of autogenous vaccines. Biomedical Science and Engineering, 1(1). https://doi.org/10.4081/bse.114

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