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...
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doaj-5dc88b1b04164c82a8b0a57be151a9ea2020-11-25T02:17:20ZengPublic 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.http://europepmc.org/articles/PMC2829072?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Dmitriy Mazurov Anna Ilinskaya Gisela Heidecker Patricia Lloyd David Derse |
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 |
author_facet |
Dmitriy Mazurov Anna Ilinskaya Gisela Heidecker Patricia Lloyd David Derse |
author_sort |
Dmitriy Mazurov |
title |
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_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_sort |
quantitative comparison of htlv-1 and hiv-1 cell-to-cell infection with new replication dependent vectors. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS Pathogens |
issn |
1553-7366 1553-7374 |
publishDate |
2010-02-01 |
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. |
url |
http://europepmc.org/articles/PMC2829072?pdf=render |
work_keys_str_mv |
AT dmitriymazurov quantitativecomparisonofhtlv1andhiv1celltocellinfectionwithnewreplicationdependentvectors AT annailinskaya quantitativecomparisonofhtlv1andhiv1celltocellinfectionwithnewreplicationdependentvectors AT giselaheidecker quantitativecomparisonofhtlv1andhiv1celltocellinfectionwithnewreplicationdependentvectors AT patricialloyd quantitativecomparisonofhtlv1andhiv1celltocellinfectionwithnewreplicationdependentvectors AT davidderse quantitativecomparisonofhtlv1andhiv1celltocellinfectionwithnewreplicationdependentvectors |
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