Document Detail


Modulation of porcine endogenous retroviruses (PERVs) expression in vitro by shRNA in the presence of cyclosporine A and dexamethasone.
MedLine Citation:
PMID:  23274329     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Despite the fact that the risk of Porcine Endogenous Retroviruses (PERV) infection and propagation in human recipients is extremely low, such an event cannot be completely ruled out, especially in immunosuppressed patients. Therefore, the aim of this study was to analyze the expression of PERVs in vitro in the presence of immunosuppression agents: cyclosporine A (CsA), and dexamethasone (DEX). We investigated the possible interactions between immunosuppression drugs, CsA and DEX, and the efficiency of anti-PERV RNAi.
MATERIAL/METHODS: Plasmid-based vectors expressing shRNAs against all PERV genes were constructed and analyzed. PERVs expression in cultures transfected with anti-PERV RNAi constructions and treated with CsA or DEX was analyzed by Real-Time RT-PCR, Western blot, and by the measurement of RT activity.
RESULTS: Both CsA and DEX inhibited PERVs expression in cell cultures in vitro. RNAi constructions efficiently knocked down PERV expression in Circe, and de novo PERV-infected HeLa and HEK-293 cell cultures. Pretreatment of Circe cultures with CsA or DEX increased PERVs knockdown by RNAi, but no specific interaction between the drugs and transfection efficiency was observed.
CONCLUSIONS: Our results demonstrate that cyclosporine A and dexamethasone decrease expression of PERVs in vitro. We also proved that these drugs did not synergize or antagonize RNAi-mediated knockdown of PERVs. These observations may be beneficial in immunosuppressed xenograft recipients; however, due to the controversial literature data concerning influence of immune suppression on graft recipients, our results should be further analyzed.
Authors:
Daniel Sypniewski; Ilona Bednarek; Daria Matczyńska; Sabina Gałka; Tomasz Loch; Dagna Sołtysik; Grzegorz Machnik; Ewa Nowak
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Annals of transplantation : quarterly of the Polish Transplantation Society     Volume:  17     ISSN:  2329-0358     ISO Abbreviation:  Ann. Transplant.     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-31     Completed Date:  2013-05-28     Revised Date:  2013-06-17    
Medline Journal Info:
Nlm Unique ID:  9802544     Medline TA:  Ann Transplant     Country:  Poland    
Other Details:
Languages:  eng     Pagination:  92-107     Citation Subset:  IM    
Affiliation:
Department of Biotechnology and Genetic Engineering, Medical University of Silesia, Sosnowiec, Poland. dsypniewski@sum.edu.pl
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Western
Cyclosporine / pharmacology*
DNA, Viral / analysis
Dexamethasone / pharmacology*
Endogenous Retroviruses / drug effects*,  genetics
Gene Expression Regulation, Viral / drug effects*
HEK293 Cells
HeLa Cells
Humans
Immunosuppressive Agents / pharmacology*
RNA Interference / drug effects*
RNA, Small Interfering / drug effects,  pharmacology*
RNA, Viral / analysis
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Swine
Transplantation, Heterologous
Chemical
Reg. No./Substance:
0/DNA, Viral; 0/Immunosuppressive Agents; 0/RNA, Small Interfering; 0/RNA, Viral; 50-02-2/Dexamethasone; 59865-13-3/Cyclosporine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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