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The superior role of coagulation factor FX over FVII in adenoviral-mediated innate immune induction of the hepatocyte: an in vitro experiment
AIM OF THE STUDY: To better understanding the contribution of coagulation factors to the extent of adenovirus-mediated innate toxicity on the hepatocyte. MATERIAL AND METHODS: Adenovirus-36 (AD) and adenovector type 5-GFP (Ad5-GFP) were propagated and titered; then, they were loaded with coagulation...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Termedia Publishing House
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7592097/ https://www.ncbi.nlm.nih.gov/pubmed/33145426 http://dx.doi.org/10.5114/ceh.2020.99512 |
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author | Ghahestani, Saeed Firoozi Shiri, Alireza Moattari, Afagh Sarvari, Jamal Tamaddon, Ali Moahammad Hossein, Seyed Younes |
author_facet | Ghahestani, Saeed Firoozi Shiri, Alireza Moattari, Afagh Sarvari, Jamal Tamaddon, Ali Moahammad Hossein, Seyed Younes |
author_sort | Ghahestani, Saeed Firoozi |
collection | PubMed |
description | AIM OF THE STUDY: To better understanding the contribution of coagulation factors to the extent of adenovirus-mediated innate toxicity on the hepatocyte. MATERIAL AND METHODS: Adenovirus-36 (AD) and adenovector type 5-GFP (Ad5-GFP) were propagated and titered; then, they were loaded with coagulation factors VII or X. The complex of adenovirus with coagulation factor VII and X were for size and charge parameters. After adding AD-VII and AD-X complexes, the expression levels of innate inflammatory genes including protein kinase R (PKR), interleukin (IL)-1β, IL-8 and IL-18 were measured by Real-time PCR on a hepatocellular carcinoma cell line, HepG2. RESULTS: The loading of coagulation factors VII and X on Ad5-GFP enhanced the transduction rate up to 50% and 60% (p < 0.05), respectively, compared to the adenovector alone (30%) (p < 0.05). The formation of the coagulation factor-virus complex leads to multimodal size distribution with an increase in average hydrodynamic size and absolute zeta potential. The qPCR results showed that PKR expression increased significantly after treatment with all adenoviruses. These findings also showed that AD had a significant (p = 0.0152) inflammatory impact on Hep-G2. However, AD which was loaded with FX (AD-X) exhibited the most inflammatory effect (p = 0.0164). Significantly, the expression of IL-1β (p = 0.0041), IL-8 (p = 0.0107) and IL-18 (p = 0.0193) were also enhanced following FX loading. On the other hand, the AD-VII complex showed the least effect of innate immune induction when compared to the negative control (p < 0.05). CONCLUSIONS: The loading of coagulation factors, particularly FX, could enhance the transduction efficiency of Ad5-GFP. Furthermore, adenovirus loaded with FX exhibited more innate toxicity on the hepatocytes, while it was not the case for FVII. |
format | Online Article Text |
id | pubmed-7592097 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Termedia Publishing House |
record_format | MEDLINE/PubMed |
spelling | pubmed-75920972020-11-02 The superior role of coagulation factor FX over FVII in adenoviral-mediated innate immune induction of the hepatocyte: an in vitro experiment Ghahestani, Saeed Firoozi Shiri, Alireza Moattari, Afagh Sarvari, Jamal Tamaddon, Ali Moahammad Hossein, Seyed Younes Clin Exp Hepatol Original Paper AIM OF THE STUDY: To better understanding the contribution of coagulation factors to the extent of adenovirus-mediated innate toxicity on the hepatocyte. MATERIAL AND METHODS: Adenovirus-36 (AD) and adenovector type 5-GFP (Ad5-GFP) were propagated and titered; then, they were loaded with coagulation factors VII or X. The complex of adenovirus with coagulation factor VII and X were for size and charge parameters. After adding AD-VII and AD-X complexes, the expression levels of innate inflammatory genes including protein kinase R (PKR), interleukin (IL)-1β, IL-8 and IL-18 were measured by Real-time PCR on a hepatocellular carcinoma cell line, HepG2. RESULTS: The loading of coagulation factors VII and X on Ad5-GFP enhanced the transduction rate up to 50% and 60% (p < 0.05), respectively, compared to the adenovector alone (30%) (p < 0.05). The formation of the coagulation factor-virus complex leads to multimodal size distribution with an increase in average hydrodynamic size and absolute zeta potential. The qPCR results showed that PKR expression increased significantly after treatment with all adenoviruses. These findings also showed that AD had a significant (p = 0.0152) inflammatory impact on Hep-G2. However, AD which was loaded with FX (AD-X) exhibited the most inflammatory effect (p = 0.0164). Significantly, the expression of IL-1β (p = 0.0041), IL-8 (p = 0.0107) and IL-18 (p = 0.0193) were also enhanced following FX loading. On the other hand, the AD-VII complex showed the least effect of innate immune induction when compared to the negative control (p < 0.05). CONCLUSIONS: The loading of coagulation factors, particularly FX, could enhance the transduction efficiency of Ad5-GFP. Furthermore, adenovirus loaded with FX exhibited more innate toxicity on the hepatocytes, while it was not the case for FVII. Termedia Publishing House 2020-09-30 2020-09 /pmc/articles/PMC7592097/ /pubmed/33145426 http://dx.doi.org/10.5114/ceh.2020.99512 Text en Copyright © 2020 Clinical and Experimental Hepatology http://creativecommons.org/licenses/by-nc-sa/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0). License (http://creativecommons.org/licenses/by-nc-sa/4.0/) |
spellingShingle | Original Paper Ghahestani, Saeed Firoozi Shiri, Alireza Moattari, Afagh Sarvari, Jamal Tamaddon, Ali Moahammad Hossein, Seyed Younes The superior role of coagulation factor FX over FVII in adenoviral-mediated innate immune induction of the hepatocyte: an in vitro experiment |
title | The superior role of coagulation factor FX over FVII in adenoviral-mediated innate immune induction of the hepatocyte: an in vitro experiment |
title_full | The superior role of coagulation factor FX over FVII in adenoviral-mediated innate immune induction of the hepatocyte: an in vitro experiment |
title_fullStr | The superior role of coagulation factor FX over FVII in adenoviral-mediated innate immune induction of the hepatocyte: an in vitro experiment |
title_full_unstemmed | The superior role of coagulation factor FX over FVII in adenoviral-mediated innate immune induction of the hepatocyte: an in vitro experiment |
title_short | The superior role of coagulation factor FX over FVII in adenoviral-mediated innate immune induction of the hepatocyte: an in vitro experiment |
title_sort | superior role of coagulation factor fx over fvii in adenoviral-mediated innate immune induction of the hepatocyte: an in vitro experiment |
topic | Original Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7592097/ https://www.ncbi.nlm.nih.gov/pubmed/33145426 http://dx.doi.org/10.5114/ceh.2020.99512 |
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