The Deleterious Effects of Shiga Toxin Type 2 Are Neutralized In Vitro by FabF8:Stx2 Recombinant Monoclonal Antibody

Hemolytic Uremic Syndrome (HUS) associated with Shiga-toxigenic <i>Escherichia coli</i> (STEC) infections is the principal cause of acute renal injury in pediatric age groups. Shiga toxin type 2 (Stx2) has in vitro cytotoxic effects on kidney cells, including human glomerular endothelial...

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Autores principales:	Daniela Luz, Fernando D. Gómez, Raíssa L. Ferreira, Bruna S. Melo, Beatriz E. C. Guth, Wagner Quintilio, Ana Maria Moro, Agostina Presta, Flavia Sacerdoti, Cristina Ibarra, Gang Chen, Sachdev S. Sidhu, María Marta Amaral, Roxane M. F. Piazza
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	STEC Stx2 antibody fragment monoclonal antibody Medicine R
Acceso en línea:	https://doaj.org/article/ee4e3434a60a4688a020358caa726ff9
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spelling	oai:doaj.org-article:ee4e3434a60a4688a020358caa726ff92021-11-25T19:09:09ZThe Deleterious Effects of Shiga Toxin Type 2 Are Neutralized In Vitro by FabF8:Stx2 Recombinant Monoclonal Antibody10.3390/toxins131108252072-6651https://doaj.org/article/ee4e3434a60a4688a020358caa726ff92021-11-01T00:00:00Zhttps://www.mdpi.com/2072-6651/13/11/825https://doaj.org/toc/2072-6651Hemolytic Uremic Syndrome (HUS) associated with Shiga-toxigenic <i>Escherichia coli</i> (STEC) infections is the principal cause of acute renal injury in pediatric age groups. Shiga toxin type 2 (Stx2) has in vitro cytotoxic effects on kidney cells, including human glomerular endothelial (HGEC) and Vero cells. Neither a licensed vaccine nor effective therapy for HUS is available for humans. Recombinant antibodies against Stx2, produced in bacteria, appeared as the utmost tool to prevent HUS. Therefore, in this work, a recombinant FabF8:Stx2 was selected from a human Fab antibody library by phage display, characterized, and analyzed for its ability to neutralize the Stx activity from different STEC-Stx2 and Stx1/Stx2 producing strains in a gold standard Vero cell assay, and the Stx2 cytotoxic effects on primary cultures of HGEC. This recombinant Fab showed a dissociation constant of 13.8 nM and a half maximum effective concentration (EC<sub>50</sub>) of 160 ng/mL to Stx2. Additionally, FabF8:Stx2 neutralized, in different percentages, the cytotoxic effects of Stx2 and Stx1/2 from different STEC strains on Vero cells. Moreover, it significantly prevented the deleterious effects of Stx2 in a dose-dependent manner (up to 83%) in HGEC and protected this cell up to 90% from apoptosis and necrosis. Therefore, this novel and simple anti-Stx2 biomolecule will allow further investigation as a new therapeutic option that could improve STEC and HUS patient outcomes.Daniela LuzFernando D. GómezRaíssa L. FerreiraBruna S. MeloBeatriz E. C. GuthWagner QuintilioAna Maria MoroAgostina PrestaFlavia SacerdotiCristina IbarraGang ChenSachdev S. SidhuMaría Marta AmaralRoxane M. F. PiazzaMDPI AGarticleSTECStx2antibody fragmentmonoclonal antibodyMedicineRENToxins, Vol 13, Iss 825, p 825 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	STEC Stx2 antibody fragment monoclonal antibody Medicine R
spellingShingle	STEC Stx2 antibody fragment monoclonal antibody Medicine R Daniela Luz Fernando D. Gómez Raíssa L. Ferreira Bruna S. Melo Beatriz E. C. Guth Wagner Quintilio Ana Maria Moro Agostina Presta Flavia Sacerdoti Cristina Ibarra Gang Chen Sachdev S. Sidhu María Marta Amaral Roxane M. F. Piazza The Deleterious Effects of Shiga Toxin Type 2 Are Neutralized In Vitro by FabF8:Stx2 Recombinant Monoclonal Antibody
description	Hemolytic Uremic Syndrome (HUS) associated with Shiga-toxigenic <i>Escherichia coli</i> (STEC) infections is the principal cause of acute renal injury in pediatric age groups. Shiga toxin type 2 (Stx2) has in vitro cytotoxic effects on kidney cells, including human glomerular endothelial (HGEC) and Vero cells. Neither a licensed vaccine nor effective therapy for HUS is available for humans. Recombinant antibodies against Stx2, produced in bacteria, appeared as the utmost tool to prevent HUS. Therefore, in this work, a recombinant FabF8:Stx2 was selected from a human Fab antibody library by phage display, characterized, and analyzed for its ability to neutralize the Stx activity from different STEC-Stx2 and Stx1/Stx2 producing strains in a gold standard Vero cell assay, and the Stx2 cytotoxic effects on primary cultures of HGEC. This recombinant Fab showed a dissociation constant of 13.8 nM and a half maximum effective concentration (EC<sub>50</sub>) of 160 ng/mL to Stx2. Additionally, FabF8:Stx2 neutralized, in different percentages, the cytotoxic effects of Stx2 and Stx1/2 from different STEC strains on Vero cells. Moreover, it significantly prevented the deleterious effects of Stx2 in a dose-dependent manner (up to 83%) in HGEC and protected this cell up to 90% from apoptosis and necrosis. Therefore, this novel and simple anti-Stx2 biomolecule will allow further investigation as a new therapeutic option that could improve STEC and HUS patient outcomes.
format	article
author	Daniela Luz Fernando D. Gómez Raíssa L. Ferreira Bruna S. Melo Beatriz E. C. Guth Wagner Quintilio Ana Maria Moro Agostina Presta Flavia Sacerdoti Cristina Ibarra Gang Chen Sachdev S. Sidhu María Marta Amaral Roxane M. F. Piazza
author_facet	Daniela Luz Fernando D. Gómez Raíssa L. Ferreira Bruna S. Melo Beatriz E. C. Guth Wagner Quintilio Ana Maria Moro Agostina Presta Flavia Sacerdoti Cristina Ibarra Gang Chen Sachdev S. Sidhu María Marta Amaral Roxane M. F. Piazza
author_sort	Daniela Luz
title	The Deleterious Effects of Shiga Toxin Type 2 Are Neutralized In Vitro by FabF8:Stx2 Recombinant Monoclonal Antibody
title_short	The Deleterious Effects of Shiga Toxin Type 2 Are Neutralized In Vitro by FabF8:Stx2 Recombinant Monoclonal Antibody
title_full	The Deleterious Effects of Shiga Toxin Type 2 Are Neutralized In Vitro by FabF8:Stx2 Recombinant Monoclonal Antibody
title_fullStr	The Deleterious Effects of Shiga Toxin Type 2 Are Neutralized In Vitro by FabF8:Stx2 Recombinant Monoclonal Antibody
title_full_unstemmed	The Deleterious Effects of Shiga Toxin Type 2 Are Neutralized In Vitro by FabF8:Stx2 Recombinant Monoclonal Antibody
title_sort	deleterious effects of shiga toxin type 2 are neutralized in vitro by fabf8:stx2 recombinant monoclonal antibody
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/ee4e3434a60a4688a020358caa726ff9
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The Deleterious Effects of Shiga Toxin Type 2 Are Neutralized In Vitro by FabF8:Stx2 Recombinant Monoclonal Antibody

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