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Complement system activation contributes to the ependymal damage induced by microbial neuraminidase

BACKGROUND: In the rat brain, a single intracerebroventricular injection of neuraminidase from Clostridium perfringens induces ependymal detachment and death. This injury occurs before the infiltration of inflammatory blood cells; some reports implicate the complement system as a cause of these inju...

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Autores principales: Granados-Durán, Pablo, López-Ávalos, María Dolores, Hughes, Timothy R., Johnson, Krista, Morgan, B. Paul, Tamburini, Paul P., Fernández-Llebrez, Pedro, Grondona, Jesús M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4875702/
https://www.ncbi.nlm.nih.gov/pubmed/27209022
http://dx.doi.org/10.1186/s12974-016-0576-9
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author Granados-Durán, Pablo
López-Ávalos, María Dolores
Hughes, Timothy R.
Johnson, Krista
Morgan, B. Paul
Tamburini, Paul P.
Fernández-Llebrez, Pedro
Grondona, Jesús M.
author_facet Granados-Durán, Pablo
López-Ávalos, María Dolores
Hughes, Timothy R.
Johnson, Krista
Morgan, B. Paul
Tamburini, Paul P.
Fernández-Llebrez, Pedro
Grondona, Jesús M.
author_sort Granados-Durán, Pablo
collection PubMed
description BACKGROUND: In the rat brain, a single intracerebroventricular injection of neuraminidase from Clostridium perfringens induces ependymal detachment and death. This injury occurs before the infiltration of inflammatory blood cells; some reports implicate the complement system as a cause of these injuries. Here, we set out to test the role of complement. METHODS: The assembly of the complement membrane attack complex on the ependymal epithelium of rats injected with neuraminidase was analyzed by immunohistochemistry. Complement activation, triggered by neuraminidase, and the participation of different activation pathways were analyzed by Western blot. In vitro studies used primary cultures of ependymal cells and explants of the septal ventricular wall. In these models, ependymal cells were exposed to neuraminidase in the presence or absence of complement, and their viability was assessed by observing beating of cilia or by trypan blue staining. The role of complement in ependymal damage induced by neuraminidase was analyzed in vivo in two rat models of complement blockade: systemic inhibition of C5 by using a function blocking antibody and testing in C6-deficient rats. RESULTS: The complement membrane attack complex immunolocalized on the ependymal surface in rats injected intracerebroventricularly with neuraminidase. C3 activation fragments were found in serum and cerebrospinal fluid of rats treated with neuraminidase, suggesting that neuraminidase itself activates complement. In ventricular wall explants and isolated ependymal cells, treatment with neuraminidase alone induced ependymal cell death; however, the addition of complement caused increased cell death and disorganization of the ependymal epithelium. In rats treated with anti-C5 and in C6-deficient rats, intracerebroventricular injection of neuraminidase provoked reduced ependymal alterations compared to non-treated or control rats. Immunohistochemistry confirmed the absence of membrane attack complex on the ependymal surfaces of neuraminidase-exposed rats treated with anti-C5 or deficient in C6. CONCLUSIONS: These results demonstrate that the complement system contributes to ependymal damage and death caused by neuraminidase. However, neuraminidase alone can induce moderate ependymal damage without the aid of complement.
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spelling pubmed-48757022016-05-22 Complement system activation contributes to the ependymal damage induced by microbial neuraminidase Granados-Durán, Pablo López-Ávalos, María Dolores Hughes, Timothy R. Johnson, Krista Morgan, B. Paul Tamburini, Paul P. Fernández-Llebrez, Pedro Grondona, Jesús M. J Neuroinflammation Research BACKGROUND: In the rat brain, a single intracerebroventricular injection of neuraminidase from Clostridium perfringens induces ependymal detachment and death. This injury occurs before the infiltration of inflammatory blood cells; some reports implicate the complement system as a cause of these injuries. Here, we set out to test the role of complement. METHODS: The assembly of the complement membrane attack complex on the ependymal epithelium of rats injected with neuraminidase was analyzed by immunohistochemistry. Complement activation, triggered by neuraminidase, and the participation of different activation pathways were analyzed by Western blot. In vitro studies used primary cultures of ependymal cells and explants of the septal ventricular wall. In these models, ependymal cells were exposed to neuraminidase in the presence or absence of complement, and their viability was assessed by observing beating of cilia or by trypan blue staining. The role of complement in ependymal damage induced by neuraminidase was analyzed in vivo in two rat models of complement blockade: systemic inhibition of C5 by using a function blocking antibody and testing in C6-deficient rats. RESULTS: The complement membrane attack complex immunolocalized on the ependymal surface in rats injected intracerebroventricularly with neuraminidase. C3 activation fragments were found in serum and cerebrospinal fluid of rats treated with neuraminidase, suggesting that neuraminidase itself activates complement. In ventricular wall explants and isolated ependymal cells, treatment with neuraminidase alone induced ependymal cell death; however, the addition of complement caused increased cell death and disorganization of the ependymal epithelium. In rats treated with anti-C5 and in C6-deficient rats, intracerebroventricular injection of neuraminidase provoked reduced ependymal alterations compared to non-treated or control rats. Immunohistochemistry confirmed the absence of membrane attack complex on the ependymal surfaces of neuraminidase-exposed rats treated with anti-C5 or deficient in C6. CONCLUSIONS: These results demonstrate that the complement system contributes to ependymal damage and death caused by neuraminidase. However, neuraminidase alone can induce moderate ependymal damage without the aid of complement. BioMed Central 2016-05-21 /pmc/articles/PMC4875702/ /pubmed/27209022 http://dx.doi.org/10.1186/s12974-016-0576-9 Text en © Granados-Durán et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Granados-Durán, Pablo
López-Ávalos, María Dolores
Hughes, Timothy R.
Johnson, Krista
Morgan, B. Paul
Tamburini, Paul P.
Fernández-Llebrez, Pedro
Grondona, Jesús M.
Complement system activation contributes to the ependymal damage induced by microbial neuraminidase
title Complement system activation contributes to the ependymal damage induced by microbial neuraminidase
title_full Complement system activation contributes to the ependymal damage induced by microbial neuraminidase
title_fullStr Complement system activation contributes to the ependymal damage induced by microbial neuraminidase
title_full_unstemmed Complement system activation contributes to the ependymal damage induced by microbial neuraminidase
title_short Complement system activation contributes to the ependymal damage induced by microbial neuraminidase
title_sort complement system activation contributes to the ependymal damage induced by microbial neuraminidase
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4875702/
https://www.ncbi.nlm.nih.gov/pubmed/27209022
http://dx.doi.org/10.1186/s12974-016-0576-9
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