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Negative feedback circuit for toll like receptor-8 activation in human embryonic Kidney 293 using outer membrane vesicle delivered bi-specific siRNA

BACKGROUND: TLR8 assists in antiviral approach by producing Type 1 INF via MyD88 dependent IRF7 pathway. However, over expression of INFα/β molecule poses threat by developing tolerance in chronic infection cases and enhancing inflammatory response. Here we report a bi-specific siRNA based complex w...

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Autores principales: Adhikari, Anurag, Gupta, Birendra Prasad, Manandhar, Krishna Das, Mishra, Shravan Kumar, Saiju, Hari Krishna, Shrestha, Rajendra Maan, Mishra, Nawneet, Sharma, Shishir
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4510891/
https://www.ncbi.nlm.nih.gov/pubmed/26202481
http://dx.doi.org/10.1186/s12865-015-0109-9
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author Adhikari, Anurag
Gupta, Birendra Prasad
Manandhar, Krishna Das
Mishra, Shravan Kumar
Saiju, Hari Krishna
Shrestha, Rajendra Maan
Mishra, Nawneet
Sharma, Shishir
author_facet Adhikari, Anurag
Gupta, Birendra Prasad
Manandhar, Krishna Das
Mishra, Shravan Kumar
Saiju, Hari Krishna
Shrestha, Rajendra Maan
Mishra, Nawneet
Sharma, Shishir
author_sort Adhikari, Anurag
collection PubMed
description BACKGROUND: TLR8 assists in antiviral approach by producing Type 1 INF via MyD88 dependent IRF7 pathway. However, over expression of INFα/β molecule poses threat by developing tolerance in chronic infection cases and enhancing inflammatory response. Here we report a bi-specific siRNA based complex which differentially activates and silences the TLR8 and MYD88 respectively in a negatively regulated fashion. RESULTS: Outer membrane vesicle from Escherichia coli used for siRNA delivery was observed more efficient when attached with invasive protein Ail along with OmpA (P < 0.001) in HEK293-TLR8 cell line. siRNA complexed with p19 protein was efficient in activating TLR8, confirmed by the increment of INFβ molecules (P < 0.001) in HEK293-TLR8 compared to its counterpart. Fusion of lipid bilayer of endosomal compartment was significant at pH 4.5 when fusogenic peptides (diINF-7) were incubated in membrane vesicle, thus facilitating the escape of siRNA complex to the host cytoplasm in order to silence MyD88 transcript (P < 0.001). CONCLUSIONS: We investigated the activation of TLR8 by bi-specific si-RNA for the production of INFβ. In the same setting we showed that bi-specific si-RNA was able to silence MyD88 transcript in a delayed manner. For the cases of auto immune disease and inflammation where over activation of endosomal TLRs poses serious threat, bi specific siRNA could be used as negative feedback controlled system. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12865-015-0109-9) contains supplementary material, which is available to authorized users.
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spelling pubmed-45108912015-07-23 Negative feedback circuit for toll like receptor-8 activation in human embryonic Kidney 293 using outer membrane vesicle delivered bi-specific siRNA Adhikari, Anurag Gupta, Birendra Prasad Manandhar, Krishna Das Mishra, Shravan Kumar Saiju, Hari Krishna Shrestha, Rajendra Maan Mishra, Nawneet Sharma, Shishir BMC Immunol Research Article BACKGROUND: TLR8 assists in antiviral approach by producing Type 1 INF via MyD88 dependent IRF7 pathway. However, over expression of INFα/β molecule poses threat by developing tolerance in chronic infection cases and enhancing inflammatory response. Here we report a bi-specific siRNA based complex which differentially activates and silences the TLR8 and MYD88 respectively in a negatively regulated fashion. RESULTS: Outer membrane vesicle from Escherichia coli used for siRNA delivery was observed more efficient when attached with invasive protein Ail along with OmpA (P < 0.001) in HEK293-TLR8 cell line. siRNA complexed with p19 protein was efficient in activating TLR8, confirmed by the increment of INFβ molecules (P < 0.001) in HEK293-TLR8 compared to its counterpart. Fusion of lipid bilayer of endosomal compartment was significant at pH 4.5 when fusogenic peptides (diINF-7) were incubated in membrane vesicle, thus facilitating the escape of siRNA complex to the host cytoplasm in order to silence MyD88 transcript (P < 0.001). CONCLUSIONS: We investigated the activation of TLR8 by bi-specific si-RNA for the production of INFβ. In the same setting we showed that bi-specific si-RNA was able to silence MyD88 transcript in a delayed manner. For the cases of auto immune disease and inflammation where over activation of endosomal TLRs poses serious threat, bi specific siRNA could be used as negative feedback controlled system. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12865-015-0109-9) contains supplementary material, which is available to authorized users. BioMed Central 2015-07-23 /pmc/articles/PMC4510891/ /pubmed/26202481 http://dx.doi.org/10.1186/s12865-015-0109-9 Text en © Adhikari et al. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. 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 Article
Adhikari, Anurag
Gupta, Birendra Prasad
Manandhar, Krishna Das
Mishra, Shravan Kumar
Saiju, Hari Krishna
Shrestha, Rajendra Maan
Mishra, Nawneet
Sharma, Shishir
Negative feedback circuit for toll like receptor-8 activation in human embryonic Kidney 293 using outer membrane vesicle delivered bi-specific siRNA
title Negative feedback circuit for toll like receptor-8 activation in human embryonic Kidney 293 using outer membrane vesicle delivered bi-specific siRNA
title_full Negative feedback circuit for toll like receptor-8 activation in human embryonic Kidney 293 using outer membrane vesicle delivered bi-specific siRNA
title_fullStr Negative feedback circuit for toll like receptor-8 activation in human embryonic Kidney 293 using outer membrane vesicle delivered bi-specific siRNA
title_full_unstemmed Negative feedback circuit for toll like receptor-8 activation in human embryonic Kidney 293 using outer membrane vesicle delivered bi-specific siRNA
title_short Negative feedback circuit for toll like receptor-8 activation in human embryonic Kidney 293 using outer membrane vesicle delivered bi-specific siRNA
title_sort negative feedback circuit for toll like receptor-8 activation in human embryonic kidney 293 using outer membrane vesicle delivered bi-specific sirna
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4510891/
https://www.ncbi.nlm.nih.gov/pubmed/26202481
http://dx.doi.org/10.1186/s12865-015-0109-9
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