Optimization of lentiviral vector transduction into peripheral blood mononuclear cells in combination with the fibronectin fragment CH-296 stimulation

Large scale T-cell expansion and efficient gene transduction are required for adoptive T-cell gene therapy. Based on our previous observations, human peripheral blood mononuclear cells (PBMCs) can be expanded efficiently while conserving a naïve phenotype by stimulating with both recombinant human f...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 2011-03, Vol.149 (3), p.285-292
Main Authors: Chono, Hideto, Goto, Yumi, Yamakawa, Satoko, Tanaka, Shinya, Tosaka, Yasuhiro, Nukaya, Ikuei, Mineno, Junichi
Format: Article
Language:English
Subjects:
Antibodies, Monoclonal - immunology
Antibodies, Monoclonal - pharmacology
CD28 Antigens - immunology
CD3 Complex - immunology
Fibronectins - chemistry
Fibronectins - pharmacology
Flow Cytometry
Genetic Vectors - genetics
Humans
Immunotherapy, Adoptive
Lentivirus - genetics
Leukocytes, Mononuclear - cytology
Leukocytes, Mononuclear - drug effects
Leukocytes, Mononuclear - metabolism
Lymphocyte Activation - drug effects
T-Lymphocytes - cytology
T-Lymphocytes - drug effects
T-Lymphocytes - metabolism
Transduction, Genetic - methods
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Summary:Large scale T-cell expansion and efficient gene transduction are required for adoptive T-cell gene therapy. Based on our previous observations, human peripheral blood mononuclear cells (PBMCs) can be expanded efficiently while conserving a naïve phenotype by stimulating with both recombinant human fibronectin fragment (CH-296) and anti-CD3 monoclonal antibodies. In this article, we explored the possibility of using this co-stimulation method to generate engineered T cells using lentiviral vector. Human PBMCs were stimulated with anti-CD3 together with immobilized CH-296 or anti-CD28 antibody as well as anti-CD3/anti-CD28 conjugated beads and transduced with lentiviral vector simultaneously. Co-stimulation with CH-296 gave superior transduction efficiency than with anti-CD28. Next, PBMCs were stimulated and transduced with anti-CD3/CH-296 or with anti-CD3/CD28 beads. T-cell expansion, gene transfer efficiencies and immunophenotypes were analysed. Stimulation with anti-CD3/CH-296 resulted in more than 10-times higher cell expansion and higher gene transfer efficiency with conservation of the naïve phenotype compared with anti-CD3/CD28 stimulation method. Thus, lentiviral transduction with anti-CD3/CH-296 co-stimulation is an efficient way to generate large numbers of genetically modified T cells and may be suitable for many gene therapy protocols that use adoptive T-cell transfer therapy.
ISSN:0021-924X
1756-2651
DOI:10.1093/jb/mvq135