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Recombinant Retroviruses Pseudotyped With the Vesicular Stomatitis Virus G Glycoprotein Mediate Both Stable Gene Transfer and Pseudotransduction in Human Peripheral Blood Lymphocytes
H.F. Gallardo,
C. Tan,
D. Ory, and
M. Sadelain
From the Department of Human Genetics, Memorial Sloan-Kettering Cancer Center, New York, NY; the Department of Medicine, Cardiovascular Division, Washington University School of Medicine, St Louis, MO; the Immunology Program, Sloan-Kettering Institute, New York, NY; and the Graduate School of Medical Sciences, Cornell University Medical College, New York, NY.
It is essential for the study of T-cell function and the improvement of adoptive cell therapies to efficiently generate large populations of human primary T cells that reliably express foreign genes. This goal is achieved by using recombinant retroviruses pseudotyped with either the gibbon ape leukemia virus (GaLV) envelope or the vesicular stomatitis virus G (VSV-G) glycoprotein. We show here that both retroviral particles mediate stable gene transfer in CD4+ and in CD8+ peripheral blood lymphocytes cultured under optimized conditions. However, VSV-G-pseudotyped virions may cause transduction artifacts that must be carefully excluded. The VSV-G virions require 10- to 100-fold higher concentrations of infectious particles to achieve levels of gene transfer comparable to GaLV-virions. Nonetheless, the physical stability of VSV-G-coated particles enables the concentration of viral stocks to 109 infectious particles per milliliter or more.
Blood, Vol. 90 No. 3 (August 1), 1997:
pp. 952-957
© 1997 by The American Society of Hematology.
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