Human Serum Albumin Is an Essential Component of the Host Defense Mechanism Against Clostridium difficile Intoxication

Alessandra di Masi, Loris Leboffe, Fabio Polticelli, Federica Tonon, Cristina Zennaro, Marianna Caterino, Pasquale Stano, Stephan Fischer, Marlen Hägele, Martin Müller, Alexander Kleger, Panagiotis Papatheodorou, Giuseppina Nocca, Alessandro Arcovito, Andrea Gori, Margherita Ruoppolo, Holger Barth, Nicola Petrosillo, Paolo Ascenzi, Stefano Di Bella

Research output: Contribution to journalArticle

Abstract

Background: The pathogenic effects of Clostridium difficile are primarily attributable to the production of the large protein toxins (C difficile toxins [Tcd]) A (TcdA) and B (TcdB). These toxins monoglucosylate Rho GTPases in the cytosol of host cells, causing destruction of the actin cytoskeleton with cytotoxic effects. Low human serum albumin (HSA) levels indicate a higher risk of acquiring and developing a severe C difficile infection (CDI) and are associated with recurrent and fatal disease.

Methods: We used a combined approach based on docking simulation and biochemical analyses that were performed in vitro on purified proteins and in human epithelial colorectal adenocarcinoma cells (Caco-2), and in vivo on stem cell-derived human intestinal organoids and zebrafish embryos.

Results: Our results show that HSA specifically binds via its domain II to TcdA and TcdB and thereby induces their autoproteolytic cleavage at physiological concentrations. This process impairs toxin internalization into the host cells and reduces the toxin-dependent glucosylation of Rho proteins.

Conclusions: Our data provide evidence for a specific HSA-dependent self-defense mechanism against C difficile toxins and provide an explanation for the clinical correlation between CDI severity and hypoalbuminemia.

Original languageEnglish
Pages (from-to)1424-1435
Number of pages12
JournalThe Journal of infectious diseases
Volume218
Issue number9
DOIs
Publication statusPublished - Sep 22 2018

Fingerprint

Clostridium difficile
Serum Albumin
Organoids
Hypoalbuminemia
rho GTP-Binding Proteins
Caco-2 Cells
Zebrafish
Protein C
Infection
Actin Cytoskeleton
Cytosol
Proteins
Adenocarcinoma
Stem Cells
Embryonic Structures

Cite this

di Masi, A., Leboffe, L., Polticelli, F., Tonon, F., Zennaro, C., Caterino, M., ... Di Bella, S. (2018). Human Serum Albumin Is an Essential Component of the Host Defense Mechanism Against Clostridium difficile Intoxication. The Journal of infectious diseases, 218(9), 1424-1435. https://doi.org/10.1093/infdis/jiy338

Human Serum Albumin Is an Essential Component of the Host Defense Mechanism Against Clostridium difficile Intoxication. / di Masi, Alessandra; Leboffe, Loris; Polticelli, Fabio; Tonon, Federica; Zennaro, Cristina; Caterino, Marianna; Stano, Pasquale; Fischer, Stephan; Hägele, Marlen; Müller, Martin; Kleger, Alexander; Papatheodorou, Panagiotis; Nocca, Giuseppina; Arcovito, Alessandro; Gori, Andrea; Ruoppolo, Margherita; Barth, Holger; Petrosillo, Nicola; Ascenzi, Paolo; Di Bella, Stefano.

In: The Journal of infectious diseases, Vol. 218, No. 9, 22.09.2018, p. 1424-1435.

Research output: Contribution to journalArticle

di Masi, A, Leboffe, L, Polticelli, F, Tonon, F, Zennaro, C, Caterino, M, Stano, P, Fischer, S, Hägele, M, Müller, M, Kleger, A, Papatheodorou, P, Nocca, G, Arcovito, A, Gori, A, Ruoppolo, M, Barth, H, Petrosillo, N, Ascenzi, P & Di Bella, S 2018, 'Human Serum Albumin Is an Essential Component of the Host Defense Mechanism Against Clostridium difficile Intoxication', The Journal of infectious diseases, vol. 218, no. 9, pp. 1424-1435. https://doi.org/10.1093/infdis/jiy338
di Masi, Alessandra ; Leboffe, Loris ; Polticelli, Fabio ; Tonon, Federica ; Zennaro, Cristina ; Caterino, Marianna ; Stano, Pasquale ; Fischer, Stephan ; Hägele, Marlen ; Müller, Martin ; Kleger, Alexander ; Papatheodorou, Panagiotis ; Nocca, Giuseppina ; Arcovito, Alessandro ; Gori, Andrea ; Ruoppolo, Margherita ; Barth, Holger ; Petrosillo, Nicola ; Ascenzi, Paolo ; Di Bella, Stefano. / Human Serum Albumin Is an Essential Component of the Host Defense Mechanism Against Clostridium difficile Intoxication. In: The Journal of infectious diseases. 2018 ; Vol. 218, No. 9. pp. 1424-1435.
@article{cc50ce84e0584ec0b0a06ee408405efb,
title = "Human Serum Albumin Is an Essential Component of the Host Defense Mechanism Against Clostridium difficile Intoxication",
abstract = "Background: The pathogenic effects of Clostridium difficile are primarily attributable to the production of the large protein toxins (C difficile toxins [Tcd]) A (TcdA) and B (TcdB). These toxins monoglucosylate Rho GTPases in the cytosol of host cells, causing destruction of the actin cytoskeleton with cytotoxic effects. Low human serum albumin (HSA) levels indicate a higher risk of acquiring and developing a severe C difficile infection (CDI) and are associated with recurrent and fatal disease.Methods: We used a combined approach based on docking simulation and biochemical analyses that were performed in vitro on purified proteins and in human epithelial colorectal adenocarcinoma cells (Caco-2), and in vivo on stem cell-derived human intestinal organoids and zebrafish embryos.Results: Our results show that HSA specifically binds via its domain II to TcdA and TcdB and thereby induces their autoproteolytic cleavage at physiological concentrations. This process impairs toxin internalization into the host cells and reduces the toxin-dependent glucosylation of Rho proteins.Conclusions: Our data provide evidence for a specific HSA-dependent self-defense mechanism against C difficile toxins and provide an explanation for the clinical correlation between CDI severity and hypoalbuminemia.",
author = "{di Masi}, Alessandra and Loris Leboffe and Fabio Polticelli and Federica Tonon and Cristina Zennaro and Marianna Caterino and Pasquale Stano and Stephan Fischer and Marlen H{\"a}gele and Martin M{\"u}ller and Alexander Kleger and Panagiotis Papatheodorou and Giuseppina Nocca and Alessandro Arcovito and Andrea Gori and Margherita Ruoppolo and Holger Barth and Nicola Petrosillo and Paolo Ascenzi and {Di Bella}, Stefano",
year = "2018",
month = "9",
day = "22",
doi = "10.1093/infdis/jiy338",
language = "English",
volume = "218",
pages = "1424--1435",
journal = "Journal of Infectious Diseases",
issn = "0022-1899",
publisher = "Oxford University Press",
number = "9",

}

TY - JOUR

T1 - Human Serum Albumin Is an Essential Component of the Host Defense Mechanism Against Clostridium difficile Intoxication

AU - di Masi, Alessandra

AU - Leboffe, Loris

AU - Polticelli, Fabio

AU - Tonon, Federica

AU - Zennaro, Cristina

AU - Caterino, Marianna

AU - Stano, Pasquale

AU - Fischer, Stephan

AU - Hägele, Marlen

AU - Müller, Martin

AU - Kleger, Alexander

AU - Papatheodorou, Panagiotis

AU - Nocca, Giuseppina

AU - Arcovito, Alessandro

AU - Gori, Andrea

AU - Ruoppolo, Margherita

AU - Barth, Holger

AU - Petrosillo, Nicola

AU - Ascenzi, Paolo

AU - Di Bella, Stefano

PY - 2018/9/22

Y1 - 2018/9/22

N2 - Background: The pathogenic effects of Clostridium difficile are primarily attributable to the production of the large protein toxins (C difficile toxins [Tcd]) A (TcdA) and B (TcdB). These toxins monoglucosylate Rho GTPases in the cytosol of host cells, causing destruction of the actin cytoskeleton with cytotoxic effects. Low human serum albumin (HSA) levels indicate a higher risk of acquiring and developing a severe C difficile infection (CDI) and are associated with recurrent and fatal disease.Methods: We used a combined approach based on docking simulation and biochemical analyses that were performed in vitro on purified proteins and in human epithelial colorectal adenocarcinoma cells (Caco-2), and in vivo on stem cell-derived human intestinal organoids and zebrafish embryos.Results: Our results show that HSA specifically binds via its domain II to TcdA and TcdB and thereby induces their autoproteolytic cleavage at physiological concentrations. This process impairs toxin internalization into the host cells and reduces the toxin-dependent glucosylation of Rho proteins.Conclusions: Our data provide evidence for a specific HSA-dependent self-defense mechanism against C difficile toxins and provide an explanation for the clinical correlation between CDI severity and hypoalbuminemia.

AB - Background: The pathogenic effects of Clostridium difficile are primarily attributable to the production of the large protein toxins (C difficile toxins [Tcd]) A (TcdA) and B (TcdB). These toxins monoglucosylate Rho GTPases in the cytosol of host cells, causing destruction of the actin cytoskeleton with cytotoxic effects. Low human serum albumin (HSA) levels indicate a higher risk of acquiring and developing a severe C difficile infection (CDI) and are associated with recurrent and fatal disease.Methods: We used a combined approach based on docking simulation and biochemical analyses that were performed in vitro on purified proteins and in human epithelial colorectal adenocarcinoma cells (Caco-2), and in vivo on stem cell-derived human intestinal organoids and zebrafish embryos.Results: Our results show that HSA specifically binds via its domain II to TcdA and TcdB and thereby induces their autoproteolytic cleavage at physiological concentrations. This process impairs toxin internalization into the host cells and reduces the toxin-dependent glucosylation of Rho proteins.Conclusions: Our data provide evidence for a specific HSA-dependent self-defense mechanism against C difficile toxins and provide an explanation for the clinical correlation between CDI severity and hypoalbuminemia.

U2 - 10.1093/infdis/jiy338

DO - 10.1093/infdis/jiy338

M3 - Article

C2 - 29868851

VL - 218

SP - 1424

EP - 1435

JO - Journal of Infectious Diseases

JF - Journal of Infectious Diseases

SN - 0022-1899

IS - 9

ER -