Activation conditions determine susceptibility of murine primary T-lymphocytes to retroviral infection Journal Article

Authors: Hagani, A. B.; Rivière, I.; Tan, C.; Krause, A.; Sadelain, M.
Article Title: Activation conditions determine susceptibility of murine primary T-lymphocytes to retroviral infection
Abstract: Background: Genetically modified T-lymphocytes are potential therapeutic agents for the treatment of various disorders. Successful retroviral infection of primary murine T-lymphocytes is a prerequisite to the study of adoptive cell therapies in a small animal model. The definition of factors controlling retroviral infection of T-lymphocytes would also be useful to better understand retroviral diseases. However, retroviral-mediated gene transfer into murine primary T-cells has remained elusive. Methods: In order to define the requirements for stable and efficient gene transfer in primary murine T-lymphocytes, we investigated factors capable of affecting retroviral infection. These include activation conditions (using mitogens or monoclonal antibodies), culture conditions (including media composition and cytokine addition), timing of viral exposure, retroviral receptor selection (ecotropic, amphotropic, or vesicular stomatitis virus G (VSV-G) glycoprotein receptor), and viral titer. Results: We show that efficient gene transfer can be achieved in murine T-lymphocytes, provided that a number of favorable conditions are met, in particular optimized T-cell activation conditions, optimal timing of infection, adequate interleukin-2 concentration and T-cell density, and a high viral titer. On a particulate basis, we find that ecotropic particles are more effective than amphotropic or VSV-G-pseudotyped particles, and recommend the use of a specifically selected retroviral packaging cell line. CD4+ T-cells are equally or more infectible than CD8+ lymphocytes, depending on the activation conditions. The Th1 and Th2 subsets are comparably susceptible to retroviral infection, in contrast to what has been reported in some instances of human T-cell infection by human immunodeficiency virus (HIV). Conclusions: We establish conditions that enable efficient retroviral-mediated gene transfer in murine primary T-lymphocytes. T-lymphocytes are not uniformly susceptible to retroviral infection depending on the mode of T-cell activation. These findings have implications for devising approaches to the study of T-cell biology, adoptive cell therapies, and the pathophysiology of retroviral diseases in mouse models. Copyright © 1999 John Wiley & Sons, Ltd.
Keywords: genetics; t lymphocyte; t-lymphocytes; mouse; animal; cytology; animals; mice; cell survival; cells, cultured; mice, inbred c57bl; gene transfer; c57bl mouse; gene vector; virology; genetic vectors; immunology; lymphocyte activation; cell culture; cd4+ t lymphocyte; cd4-positive t-lymphocytes; gene therapy; adoptive immunotherapy; retrovirus; retroviridae; hiv; gene transfer techniques; lymphocyte subsets; lymphocyte subpopulation; t-lymphocyte; retroviral gene transfer; humans; human; male; article; th1/th2 t-cell subsets
Journal Title: Journal of Gene Medicine
Volume: 1
Issue: 5
ISSN: 1099-498X
Publisher: John Wiley & Sons  
Date Published: 1999-09-01
Start Page: 341
End Page: 351
Language: English
PUBMED: 10738551
PROVIDER: scopus
DOI: 10.1002/(SICI)1521-2254(199909/10)1:5<341::AID-JGM58>3.0.CO;2-J
Notes: Article -- Export Date: 16 August 2016 -- Source: Scopus
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MSK Authors
  1. Michel W J Sadelain
    456 Sadelain
  2. Cuiwen   Tan
    11 Tan
  3. Anja   Krause
    6 Krause