Cargando…

RBC Adherence of Immune Complexes Containing Botulinum Toxin Improves Neutralization and Macrophage Uptake

In the paralytic disease botulism, the botulinum neurotoxin (BoNT) passes through the bloodstream to reach and inactivate neuromuscular junctions. Monoclonal antibodies (mAbs) may be useful BoNT countermeasures, as mAb combinations can rapidly clear BoNT from the blood circulation. We have previousl...

Descripción completa

Detalles Bibliográficos
Autores principales: Al-Saleem, Fetweh H., Sharma, Rashmi, Puligedda, Rama Devudu, Elias, Md., Kattala, Chandana Devi, Simon, Paul M., Simpson, Lance L., Dessain, Scott K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5450721/
https://www.ncbi.nlm.nih.gov/pubmed/28534855
http://dx.doi.org/10.3390/toxins9050173
_version_ 1783240045090570240
author Al-Saleem, Fetweh H.
Sharma, Rashmi
Puligedda, Rama Devudu
Elias, Md.
Kattala, Chandana Devi
Simon, Paul M.
Simpson, Lance L.
Dessain, Scott K.
author_facet Al-Saleem, Fetweh H.
Sharma, Rashmi
Puligedda, Rama Devudu
Elias, Md.
Kattala, Chandana Devi
Simon, Paul M.
Simpson, Lance L.
Dessain, Scott K.
author_sort Al-Saleem, Fetweh H.
collection PubMed
description In the paralytic disease botulism, the botulinum neurotoxin (BoNT) passes through the bloodstream to reach and inactivate neuromuscular junctions. Monoclonal antibodies (mAbs) may be useful BoNT countermeasures, as mAb combinations can rapidly clear BoNT from the blood circulation. We have previously shown that the BoNT-neutralizing potency of mAbs can be improved through red blood cell (RBC) immunoadherence. For example, a fusion protein (FP) that adheres biotinylated mAbs to the RBC surface enabled a pair of mAbs to neutralize 5000 LD50 BoNT/A in the mouse protection assay. Here, we added two mAbs to that combination, creating a 4-mAb:FP complex that neutralized 40,000 LD50 BoNT/A in vivo, and analyzed functional correlates of neutralization. The FP enhanced potency of BoNT/A immune complexes, providing the greatest magnitude of benefit to the 4-mAb combination. RBC binding of a BoNT/A complexed with 4-mAb:FP exhibited a bi-phasic clearance process in vivo. Most of the complexes were cleared within five minutes; the rest were cleared gradually over many hours. Peritoneal macrophages showed better uptake of the 4-mAb complex than the 3-mAb complex, and this was not affected by the presence of the FP. However, the addition of RBCs to the 4-mAb:FP BoNT/A doubled macrophage uptake of the complexes. Lastly, the 4-mAb:FP BoNT/A complex synergistically induced M2 macrophage polarization, as indicated by IL-10 expression, whether or not RBCs were present. RBC-targeted immunoadherence through the FP is a potent enhancer of mAb-mediated BoNT/A neutralization in vivo, and can have positive effects on BoNT/A sequestration, immune complex uptake, and macrophage activation.
format Online
Article
Text
id pubmed-5450721
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-54507212017-06-05 RBC Adherence of Immune Complexes Containing Botulinum Toxin Improves Neutralization and Macrophage Uptake Al-Saleem, Fetweh H. Sharma, Rashmi Puligedda, Rama Devudu Elias, Md. Kattala, Chandana Devi Simon, Paul M. Simpson, Lance L. Dessain, Scott K. Toxins (Basel) Article In the paralytic disease botulism, the botulinum neurotoxin (BoNT) passes through the bloodstream to reach and inactivate neuromuscular junctions. Monoclonal antibodies (mAbs) may be useful BoNT countermeasures, as mAb combinations can rapidly clear BoNT from the blood circulation. We have previously shown that the BoNT-neutralizing potency of mAbs can be improved through red blood cell (RBC) immunoadherence. For example, a fusion protein (FP) that adheres biotinylated mAbs to the RBC surface enabled a pair of mAbs to neutralize 5000 LD50 BoNT/A in the mouse protection assay. Here, we added two mAbs to that combination, creating a 4-mAb:FP complex that neutralized 40,000 LD50 BoNT/A in vivo, and analyzed functional correlates of neutralization. The FP enhanced potency of BoNT/A immune complexes, providing the greatest magnitude of benefit to the 4-mAb combination. RBC binding of a BoNT/A complexed with 4-mAb:FP exhibited a bi-phasic clearance process in vivo. Most of the complexes were cleared within five minutes; the rest were cleared gradually over many hours. Peritoneal macrophages showed better uptake of the 4-mAb complex than the 3-mAb complex, and this was not affected by the presence of the FP. However, the addition of RBCs to the 4-mAb:FP BoNT/A doubled macrophage uptake of the complexes. Lastly, the 4-mAb:FP BoNT/A complex synergistically induced M2 macrophage polarization, as indicated by IL-10 expression, whether or not RBCs were present. RBC-targeted immunoadherence through the FP is a potent enhancer of mAb-mediated BoNT/A neutralization in vivo, and can have positive effects on BoNT/A sequestration, immune complex uptake, and macrophage activation. MDPI 2017-05-19 /pmc/articles/PMC5450721/ /pubmed/28534855 http://dx.doi.org/10.3390/toxins9050173 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Al-Saleem, Fetweh H.
Sharma, Rashmi
Puligedda, Rama Devudu
Elias, Md.
Kattala, Chandana Devi
Simon, Paul M.
Simpson, Lance L.
Dessain, Scott K.
RBC Adherence of Immune Complexes Containing Botulinum Toxin Improves Neutralization and Macrophage Uptake
title RBC Adherence of Immune Complexes Containing Botulinum Toxin Improves Neutralization and Macrophage Uptake
title_full RBC Adherence of Immune Complexes Containing Botulinum Toxin Improves Neutralization and Macrophage Uptake
title_fullStr RBC Adherence of Immune Complexes Containing Botulinum Toxin Improves Neutralization and Macrophage Uptake
title_full_unstemmed RBC Adherence of Immune Complexes Containing Botulinum Toxin Improves Neutralization and Macrophage Uptake
title_short RBC Adherence of Immune Complexes Containing Botulinum Toxin Improves Neutralization and Macrophage Uptake
title_sort rbc adherence of immune complexes containing botulinum toxin improves neutralization and macrophage uptake
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5450721/
https://www.ncbi.nlm.nih.gov/pubmed/28534855
http://dx.doi.org/10.3390/toxins9050173
work_keys_str_mv AT alsaleemfetwehh rbcadherenceofimmunecomplexescontainingbotulinumtoxinimprovesneutralizationandmacrophageuptake
AT sharmarashmi rbcadherenceofimmunecomplexescontainingbotulinumtoxinimprovesneutralizationandmacrophageuptake
AT puligeddaramadevudu rbcadherenceofimmunecomplexescontainingbotulinumtoxinimprovesneutralizationandmacrophageuptake
AT eliasmd rbcadherenceofimmunecomplexescontainingbotulinumtoxinimprovesneutralizationandmacrophageuptake
AT kattalachandanadevi rbcadherenceofimmunecomplexescontainingbotulinumtoxinimprovesneutralizationandmacrophageuptake
AT simonpaulm rbcadherenceofimmunecomplexescontainingbotulinumtoxinimprovesneutralizationandmacrophageuptake
AT simpsonlancel rbcadherenceofimmunecomplexescontainingbotulinumtoxinimprovesneutralizationandmacrophageuptake
AT dessainscottk rbcadherenceofimmunecomplexescontainingbotulinumtoxinimprovesneutralizationandmacrophageuptake