Efficient in vitro transduction of naive murine B cells with lentiviral vectors.
The aim of this study was to determine the impact of lentiviral transduction on primary murine B cells. Studying B cell activities in vivo or using them for tolerance induction requires that the cells remain unaltered in their biological behavior except for expression of the transgene. As we show here, murine B cells can efficiently be transduced by lentiviral, VSV-G-pseudotyped vectors without the necessity of prior activation. Culture with LPS gave enhanced transduction efficiencies but led to the upregulation of CD86 and proliferation of the cells. Transduction of naive B cells by lentiviral vectors was dependent on multiplicity of infection and did not lead to a concomitant activation. Furthermore, the transduced cells could be used for studies in the NOD mouse system without altering the onset of diabetes. We conclude that lentiviral gene transfer into naive B cells is a powerful tool for manipulation of B cells for therapeutic applications.
Warncke
Max
M
Genethor GmbH, Goltzstr. 52, 10781 Berlin, Germany.
Vogt
Birgit
B
Ulrich
Jacqueline
J
von Laer
Meike Dorothee
MD
Beyer
Winfried
W
Klump
Hannes
H
Micheel
Burkhard
B
Sheriff
Ahmed
A
D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000821:virology; D002455:Cell Division; D002460:Cell Line; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins; D006801:Humans; D007108:Immune Tolerance / Q000235:genetics / Q000276:immunology; D016086:Lentivirus / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D008070:Lipopolysaccharides / Q000276:immunology / Q000494:pharmacology; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D016688:Mice, Inbred NOD; D013154:Spleen / Q000166:cytology; D014161:Transduction, Genetic / Q000379:methods
D015703:Antigens, CD; D008070:Lipopolysaccharides; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins