Recombinant retroviruses pseudotyped with the vesicular stomatitis virus G glycoprotein mediate both stable gene transfer and pseudotransduction in human peripheral blood lymphocytes.
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 10(9) infectious particles per milliliter or more.
Gallardo
H F
HF
Department of Human Genetics, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
Tan
C
C
Ory
D
D
Sadelain
M
M
D016475:3T3 Cells; D000818:Animals; D016477:Artifacts; D015496:CD4-Positive T-Lymphocytes / Q000821:virology*; D018414:CD8-Positive T-Lymphocytes / Q000821:virology*; D018076:DNA, Complementary / Q000235:genetics; D005189:False Positive Reactions; D015686:Gene Products, env / Q000502:physiology*; D005822:Genetic Vectors / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D017989:Leukemia Virus, Gibbon Ape / Q000737:chemistry / Q000502:physiology*; D008175:Lung Neoplasms / Q000473:pathology; D008562:Membrane Glycoproteins; D051379:Mice; D020800:Receptor, Nerve Growth Factor; D017475:Receptors, Nerve Growth Factor / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014161:Transduction, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured; D014721:Vesicular stomatitis Indiana virus / Q000737:chemistry / Q000502:physiology*; D014759:Viral Envelope Proteins / Q000502:physiology*
D018076:DNA, Complementary; C028391:G protein, vesicular stomatitis virus; D015686:Gene Products, env; D008562:Membrane Glycoproteins; D020800:Receptor, Nerve Growth Factor; D017475:Receptors, Nerve Growth Factor; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins