Quantitative comparison of HTLV-1 and HIV-1 cell-to-cell infection with new replication dependent vectors.

We have developed an efficient method to quantify cell-to-cell infection with single-cycle, replication dependent reporter vectors. This system was used to examine the mechanisms of infection with HTLV-1 and HIV-1 vectors in lymphocyte cell lines. Effector cells transfected with reporter vector, pac...

Full description

Saved in:
Bibliographic Details
Main Authors: Dmitriy Mazurov, Anna Ilinskaya, Gisela Heidecker, Patricia Lloyd, David Derse
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2010-02-01
Series:PLoS Pathogens
Online Access:https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1000788&type=printable
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850236156712583168
author Dmitriy Mazurov
Anna Ilinskaya
Gisela Heidecker
Patricia Lloyd
David Derse
author_facet Dmitriy Mazurov
Anna Ilinskaya
Gisela Heidecker
Patricia Lloyd
David Derse
author_sort Dmitriy Mazurov
collection DOAJ
description We have developed an efficient method to quantify cell-to-cell infection with single-cycle, replication dependent reporter vectors. This system was used to examine the mechanisms of infection with HTLV-1 and HIV-1 vectors in lymphocyte cell lines. Effector cells transfected with reporter vector, packaging vector, and Env expression plasmid produced virus-like particles that transduced reporter gene activity into cocultured target cells with zero background. Reporter gene expression was detected exclusively in target cells and required an Env-expression plasmid and a viral packaging vector, which provided essential structural and enzymatic proteins for virus replication. Cell-cell fusion did not contribute to infection, as reporter protein was rarely detected in syncytia. Coculture of transfected Jurkat T cells and target Raji/CD4 B cells enhanced HIV-1 infection two fold and HTLV-1 infection ten thousand fold in comparison with cell-free infection of Raji/CD4 cells. Agents that interfere with actin and tubulin polymerization strongly inhibited HTLV-1 and modestly decreased HIV-1 cell-to-cell infection, an indication that cytoskeletal remodeling was more important for HTLV-1 transmission. Time course studies showed that HTLV-1 transmission occurred very rapidly after cell mixing, whereas slower kinetics of HIV-1 coculture infection implies a different mechanism of infectious transmission. HTLV-1 Tax was demonstrated to play an important role in altering cell-cell interactions that enhance virus infection and replication. Interestingly, superantigen-induced synapses between Jurkat cells and Raji/CD4 cells did not enhance infection for either HTLV-1 or HIV-1. In general, the dependence on cell-to-cell infection was determined by the virus, the effector and target cell types, and by the nature of the cell-cell interaction.
format Article
id doaj-art-5dc88b1b04164c82a8b0a57be151a9ea
institution OA Journals
issn 1553-7366
1553-7374
language English
publishDate 2010-02-01
publisher Public Library of Science (PLoS)
record_format Article
series PLoS Pathogens
spelling doaj-art-5dc88b1b04164c82a8b0a57be151a9ea2025-08-20T02:02:01ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742010-02-0162e100078810.1371/journal.ppat.1000788Quantitative comparison of HTLV-1 and HIV-1 cell-to-cell infection with new replication dependent vectors.Dmitriy MazurovAnna IlinskayaGisela HeideckerPatricia LloydDavid DerseWe have developed an efficient method to quantify cell-to-cell infection with single-cycle, replication dependent reporter vectors. This system was used to examine the mechanisms of infection with HTLV-1 and HIV-1 vectors in lymphocyte cell lines. Effector cells transfected with reporter vector, packaging vector, and Env expression plasmid produced virus-like particles that transduced reporter gene activity into cocultured target cells with zero background. Reporter gene expression was detected exclusively in target cells and required an Env-expression plasmid and a viral packaging vector, which provided essential structural and enzymatic proteins for virus replication. Cell-cell fusion did not contribute to infection, as reporter protein was rarely detected in syncytia. Coculture of transfected Jurkat T cells and target Raji/CD4 B cells enhanced HIV-1 infection two fold and HTLV-1 infection ten thousand fold in comparison with cell-free infection of Raji/CD4 cells. Agents that interfere with actin and tubulin polymerization strongly inhibited HTLV-1 and modestly decreased HIV-1 cell-to-cell infection, an indication that cytoskeletal remodeling was more important for HTLV-1 transmission. Time course studies showed that HTLV-1 transmission occurred very rapidly after cell mixing, whereas slower kinetics of HIV-1 coculture infection implies a different mechanism of infectious transmission. HTLV-1 Tax was demonstrated to play an important role in altering cell-cell interactions that enhance virus infection and replication. Interestingly, superantigen-induced synapses between Jurkat cells and Raji/CD4 cells did not enhance infection for either HTLV-1 or HIV-1. In general, the dependence on cell-to-cell infection was determined by the virus, the effector and target cell types, and by the nature of the cell-cell interaction.https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1000788&type=printable
spellingShingle Dmitriy Mazurov
Anna Ilinskaya
Gisela Heidecker
Patricia Lloyd
David Derse
Quantitative comparison of HTLV-1 and HIV-1 cell-to-cell infection with new replication dependent vectors.
PLoS Pathogens
title Quantitative comparison of HTLV-1 and HIV-1 cell-to-cell infection with new replication dependent vectors.
title_full Quantitative comparison of HTLV-1 and HIV-1 cell-to-cell infection with new replication dependent vectors.
title_fullStr Quantitative comparison of HTLV-1 and HIV-1 cell-to-cell infection with new replication dependent vectors.
title_full_unstemmed Quantitative comparison of HTLV-1 and HIV-1 cell-to-cell infection with new replication dependent vectors.
title_short Quantitative comparison of HTLV-1 and HIV-1 cell-to-cell infection with new replication dependent vectors.
title_sort quantitative comparison of htlv 1 and hiv 1 cell to cell infection with new replication dependent vectors
url https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1000788&type=printable
work_keys_str_mv AT dmitriymazurov quantitativecomparisonofhtlv1andhiv1celltocellinfectionwithnewreplicationdependentvectors
AT annailinskaya quantitativecomparisonofhtlv1andhiv1celltocellinfectionwithnewreplicationdependentvectors
AT giselaheidecker quantitativecomparisonofhtlv1andhiv1celltocellinfectionwithnewreplicationdependentvectors
AT patricialloyd quantitativecomparisonofhtlv1andhiv1celltocellinfectionwithnewreplicationdependentvectors
AT davidderse quantitativecomparisonofhtlv1andhiv1celltocellinfectionwithnewreplicationdependentvectors