Dynamics of HIV-1 recombination in its natural target cells

David Levy, Grace M. Aldrovandi, Olaf Kutsch, George M. Shaw

Research output: Contribution to journalArticle

Abstract

Genetic recombination is believed to assist HIV-1 diversification and escape from host immunity and antiviral therapies, yet this process remains largely unexamined within the natural target-cell populations. We developed a method for measuring HIV-1 recombination directly that employs reporter viruses bearing functional enhanced yellow fluorescent protein (YFP) and enhanced cyan fluorescent protein (CFP) genes in which recombination produces a modified GFP gene and GFP fluorescence in the infected cells. These reporter viruses allow simultaneous quantification of the dynamics of HIV-1 infection, coinfection, and recombination in cell culture and in animal models by flow-cytometric analysis. Multiround infection assays revealed that productive cellular coinfection was subject to little functional inhibition. As a result, generation of recombinants proceeded according to the square of the infection rate during HIV-1 replication in T lymphocytes and within human thymic grafts in severe combined immunodeficient (SCID)-hu (Thy/Liv) mice. These results suggest that increases in viral load may confer a compounding risk of virus escape by means of recombinational diversification. A single round of replication in T lymphocytes in culture generated an average of nine recombination events per virus, and infection of macrophages led to ≈30 cross-over events, making HIV-1 up to an order of magnitude more recombinogenic than recognized previously and demonstrating that the infected cell exerts a profound influence on the frequency of recombination.

Original languageEnglish (US)
Pages (from-to)4204-4209
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume101
Issue number12
DOIs
StatePublished - Mar 23 2004

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Genetic Recombination
HIV-1
Viruses
Coinfection
T-Lymphocytes
Health Services Needs and Demand
Virus Diseases
Infection
Viral Load
Genes
HIV Infections
Antiviral Agents
Immunity
Animal Models
Cell Culture Techniques
Fluorescence
Macrophages
Transplants
Proteins

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Dynamics of HIV-1 recombination in its natural target cells. / Levy, David; Aldrovandi, Grace M.; Kutsch, Olaf; Shaw, George M.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 101, No. 12, 23.03.2004, p. 4204-4209.

Research output: Contribution to journalArticle

Levy, David ; Aldrovandi, Grace M. ; Kutsch, Olaf ; Shaw, George M. / Dynamics of HIV-1 recombination in its natural target cells. In: Proceedings of the National Academy of Sciences of the United States of America. 2004 ; Vol. 101, No. 12. pp. 4204-4209.
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