A human immunodeficiency virus type 1 pol gene-derived sequence (cPPT/CTS) increases the efficiency of transduction of human nondividing monocytes and T lymphocytes by lentiviral vectors

Hum Gene Ther. 2002 Oct 10;13(15):1793-807. doi: 10.1089/104303402760372909.

Abstract

We have investigated the capacity of two human immunodeficiency virus type 1-derived lentivectors, differing in the presence of a 118-bp pol fragment containing the cPPT/CTS element, to transduce human normal primary cells of different hematopoietic lineages. Infection of resting monocytes with a high multiplicity of infection (MOI > 10) revealed that the lentivirus carrying the pol fragment (cPPT) is effective, transducing 75% of cells compared with 36% for the no-cPPT vector. Even at low MOIs (< or =1) the cPPT vector still shows a better transduction efficiency than the no-cPPT vector. Moreover, transduction does not require dendritic cell differentiation. In contrast, infection of nonactivated T lymphocytes showed that both vectors, tested at high MOIs, can transduce a small, although measurable, percentage of cells (up to 10%), which may correspond to G(1a) "activated" cells as detected by simultaneous staining of DNA and RNA, in our cultures in the presence of medium alone. Furthermore, we show that the sole addition of interleukin 2 or interleukin 15 represents a full proliferative signal under our conditions and permits high transduction efficiency (up to 30% with the cPPT vector and 15% with the no-cPPT vector). Still higher transduction of T lymphocytes can be achieved after stimulation with phytohemagglutinin and interleukin 2 (up to 78% with the cPPT vector vs. 42% with the no-cPPT vector). Finally, both viruses do not transduce either resting or proliferating tonsillar B lymphocytes.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • B-Lymphocytes / drug effects
  • B-Lymphocytes / virology
  • Cell Division / drug effects
  • Fibroblasts
  • Genes, Reporter
  • Genes, pol*
  • Genetic Vectors / genetics*
  • Green Fluorescent Proteins
  • HIV-1 / genetics*
  • Humans
  • Interleukin-15 / pharmacology
  • Interleukin-2 / pharmacology
  • Kidney
  • Luminescent Proteins / analysis
  • Luminescent Proteins / genetics
  • Lymphoma, Follicular / pathology
  • Moloney murine leukemia virus / genetics
  • Monocytes / drug effects
  • Monocytes / virology*
  • Phytohemagglutinins / pharmacology
  • T-Lymphocytes / drug effects
  • T-Lymphocytes / virology*
  • Terminal Repeat Sequences
  • Transduction, Genetic*
  • Tumor Cells, Cultured

Substances

  • Interleukin-15
  • Interleukin-2
  • Luminescent Proteins
  • Phytohemagglutinins
  • Green Fluorescent Proteins