Document Detail


Entry kinetics and cell-cell transmission of surface-bound retroviral vector particles.
MedLine Citation:
PMID:  20440757     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Transduction with recombinant HIV-1 derived lentivirus vectors is a multi-step process initiated by surface attachment and subsequent receptor-directed uptake into the target cell. We previously reported the retention of vesicular stomatitis virus G protein pseudotyped particles on murine progenitor cells and their delayed cell-cell transfer.
METHODS: To examine the underlying mechanism in more detail, we used a combination of approaches focused on investigating the role of receptor-independent factors in modulating attachment.
RESULTS: The investigation of synchronized transduction reveals cell-type specific rates of vector particle clearance with substantial delays during particle entry into murine hematopoietic progenitor cells. The observed uptake kinetics from the surface of the 1 degrees cell correlate inversely with the magnitude of transfer to 2 degrees targets, corresponding with our initial observation of preferential cell-cell transfer in the context of brief vector exposures. We further demonstrate that vector particle entry into cells is associated with the cell-type specific abundance of extracellular matrix fibronectin. Residual particle-extracellular fibronectin matrix binding and 2 degrees transfer can be competitively disrupted by heparin exposure without affecting murine progenitor homing and repopulation.
CONCLUSIONS: Although cellular attachment factors, including fibronectin, aid gene transfer by colocalizing particles to cells and disfavoring early dissociation from targets, they also appear to stabilize particles on the cell surface. The present study highlights the inadvertent consequences for cell entry and cell-cell transfer.
Authors:
Lee S O'Neill; Amy M Skinner; Josha A Woodward; Peter Kurre
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The journal of gene medicine     Volume:  12     ISSN:  1521-2254     ISO Abbreviation:  J Gene Med     Publication Date:  2010 May 
Date Detail:
Created Date:  2010-05-04     Completed Date:  2011-03-10     Revised Date:  2011-07-28    
Medline Journal Info:
Nlm Unique ID:  9815764     Medline TA:  J Gene Med     Country:  England    
Other Details:
Languages:  eng     Pagination:  463-76     Citation Subset:  IM    
Copyright Information:
Copyright (c) 2010 John Wiley & Sons, Ltd.
Affiliation:
Department of Pediatrics, Papé Family Pediatric Research Institute, Oregon Health & Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Bone Marrow Cells / cytology,  drug effects,  virology
Cell Communication* / drug effects
Cell Line
Cell Lineage / drug effects
Cell Membrane / drug effects,  metabolism,  virology*
Extracellular Matrix / drug effects,  metabolism
Fibronectins / metabolism
Genetic Vectors / metabolism*
Green Fluorescent Proteins / metabolism
Heparin / pharmacology
Humans
Kinetics
Mice
Peptide Hydrolases / metabolism
Proviruses / drug effects,  metabolism
Retroviridae / drug effects,  metabolism*
Time Factors
Transduction, Genetic
Virion / drug effects,  metabolism*
Virus Integration / drug effects
Virus Internalization* / drug effects
Whole-Body Irradiation
Grant Support
ID/Acronym/Agency:
HL77231/HL/NHLBI NIH HHS; HL90765/HL/NHLBI NIH HHS; K08 HL077231-05/HL/NHLBI NIH HHS; R01 HL090765-02/HL/NHLBI NIH HHS; R01 HL090765-04/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Fibronectins; 147336-22-9/Green Fluorescent Proteins; 9005-49-6/Heparin; EC 3.4.-/Peptide Hydrolases
Comments/Corrections

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


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