Zebrafish embryo sensitivity test as in vivo platform to anti-Shiga toxin compound screening
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BIOTECHNOLOGY AND INDUSTRIAL MICROBIOLOGY - SHORT COMMUNICATION
Zebrafish embryo sensitivity test as in vivo platform to anti-Shiga toxin compound screening Bruna de Sousa Melo 1 & Bianca Helena Ventura Fernandes 2 & Monica Valdyrce Anjos Lopes-Ferreira 3,4 & Camila Henrique 1 & Roxane Maria Fontes Piazza 1 & Daniela Luz 1,5 Received: 3 March 2020 / Accepted: 18 May 2020 # Sociedade Brasileira de Microbiologia 2020
Abstract Shiga toxin-producing Escherichia coli (STEC) pathotype secretes two types of AB5 cytotoxins (Stx1 and Stx2), responsible for complications such as hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS) in infected patients, which could lead to sequels and death. Currently, there is no effective treatment against the cytotoxic effect of these toxins. However, in order to approve any therapy molecule, an animal experiment is required in order to evaluate the efficacy and safety of therapeutic approaches. The use of alternative small host models is growing among human infectious disease studies, particularly the vertebrate zebrafish model, since relevant results have been described for pathogen-host interaction. In this sense, the present work aimed to analyze the toxic effect of Shiga toxins in zebrafish embryo model in order to standardize this method in the future to be used as a fast, simple, and efficient methodology for the screening of therapeutic molecules. Herein, we demonstrated that the embryos were sensitive in a dose-dependent manner to both Stx toxins, with LD50 of 22 μg/mL for Stx1 and 33 μg/mL for Stx2, and the use of anti-Stx polyclonal antibody abolished the toxic effect. Therefore, this methodology can be a rapid alternative method for selecting promising compounds against Stx toxins, such as recombinant antibodies. Keywords Zebrafish . Shiga toxin . Toxicity . STEC
Introduction The common virulence factor among Shiga toxin– producing Escherichia coli (STEC) and related to hemolytic uremic syndrome (HUS) occurrence is a secreted toxin known as Shiga toxin (Stx) [1]. Lambda phage– located genes integrated into the STEC genome and Responsible Editor: Marina Baquerizo Martinez * Daniela Luz [email protected] 1
Laboratório de Bacteriologia, Instituto Butantan, São Paulo, Brazil
2
Laboratório de Controle Genético e Sanitário Animal, Unidade Zebrafish, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil
3
Laboratório Especial de Toxinologia, Instituto Butantan, São Paulo, Brazil
4
CeTICS FAPESP, São Paulo, Brazil
5
Laboratório de Monoclonais, Instituto de Ciência e Tecnologia, Universidade Federal de São Paulo, Rua Talim, 330, São José dos Campos, SP 12231-280, Brazil
transferred horizontally are responsible for encoding both toxin types (Stx1 and Stx2) and their variants [1, 2]. These toxins belong to a toxin AB5 class, which the B pentamer allows the toxin bind to Gb3 receptor at host cells, translocated the active subunit A into the cytoplasm, where it removes an adenine residue from an α-sarcin loop of rRNA 28S, which breaks the protein s
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