TY - JOUR
T1 - Lentiviral vectors with CMV or MHCII promoters administered in vivo
T2 - Immune reactivity versus persistence of expression
AU - Kimura, Takahiro
AU - Koya, Richard C.
AU - Anselmi, Laura
AU - Sternini, Catia
AU - Wang, He Jing
AU - Comin-Anduix, Begonya
AU - Prins, Robert M.
AU - Faure-Kumar, Emmanuelle
AU - Rozengurt, Nora
AU - Cui, Yan
AU - Kasahara, Noriyuki
AU - Stripecke, Renata
N1 - Funding Information:
We thank our colleagues from the University of California Los Angeles (UCLA), David Stout, Harvey Herschman, and Lily Wu, for technical advice for the imaging analyses. This work was supported by grants from the Margareth E. Early Research Trust and Stop Cancer and by National Institutes of Health (NIH) developmental grants (UCLA Center for In Vivo Imaging in Cancer Biology/2P50-CA086306-06 and UCLA-CURE/2P30-DK041301) (to R.S.). We appreciate the services of the CURE/UCLA Vector Core (NIH, 2P30DK041301), the CURE/UCLA Morphology and Imaging Core (NIH, 2P30DK041301), the UCLA Mouse Pathology Core, the JCCC/UCLA Flow Cytometry Core, and the UCLA Center for In Vivo Imaging in Cancer Biology (NIH 2P50CA086306-06). R.C.K. was supported by a UCLA/JCCC postdoctoral fellowship.
PY - 2007/7
Y1 - 2007/7
N2 - Lentiviral vectors (LVs) are potential tools for genetic vaccination. To improve the safety of LV vaccines, we evaluated the selectivity, bio-distribution, persistence of expression, and immune potency of vesicular stomatitis virus G (VSV-G)-pseudotyped vectors transcriptionally targeted to antigen presenting cells (APCs) through a major histocompatibility complex class II (MHCII) promoter. Control vectors contained the ubiquitous cytomegalovirus (CMV) promoter. Bio-distribution studies after intravenous injections of LVs expressing green fluorescent protein (GFP) or luciferase were conducted by a combination of flow cytometry, immunofluorescence, real-time quantitative polymerase chain reaction (RT-Q-PCR) and whole-body bioluminescence analyses. GFP-expressing vectors showed selective expression in MHCII+ cells of spleen and LV-CMV-GFP administration produced noticeable spleen inflammation, whereas LV-MHCII-GFP did not. Long-term optical imaging analyses of C57BL/6 mice injected with LV-CMV-LUC showed diminishing luciferase expression in the liver and spleen over time. Vaccination/boost with LV-CMV expressing the melanoma antigen tyrosinase-related protein 2 (TRP2) yielded dose-dependent antigen-specific CD8+ T-cell reactivity and high protection against B16 melanoma challenge. Unexpectedly, administration of LVs containing the MHCII promoter resulted in persistence of luciferase expression and viral integration in MHCII+ splenocytes and virtually no CD8+ T-cell responses against TRP2. These studies reveal that APC transduction by LVs could lead to immune reactivity (LV-CMV) or persistence of transgene expression (LV-MHCII), providing a relevant paradigm for vaccination and gene replacement approaches.
AB - Lentiviral vectors (LVs) are potential tools for genetic vaccination. To improve the safety of LV vaccines, we evaluated the selectivity, bio-distribution, persistence of expression, and immune potency of vesicular stomatitis virus G (VSV-G)-pseudotyped vectors transcriptionally targeted to antigen presenting cells (APCs) through a major histocompatibility complex class II (MHCII) promoter. Control vectors contained the ubiquitous cytomegalovirus (CMV) promoter. Bio-distribution studies after intravenous injections of LVs expressing green fluorescent protein (GFP) or luciferase were conducted by a combination of flow cytometry, immunofluorescence, real-time quantitative polymerase chain reaction (RT-Q-PCR) and whole-body bioluminescence analyses. GFP-expressing vectors showed selective expression in MHCII+ cells of spleen and LV-CMV-GFP administration produced noticeable spleen inflammation, whereas LV-MHCII-GFP did not. Long-term optical imaging analyses of C57BL/6 mice injected with LV-CMV-LUC showed diminishing luciferase expression in the liver and spleen over time. Vaccination/boost with LV-CMV expressing the melanoma antigen tyrosinase-related protein 2 (TRP2) yielded dose-dependent antigen-specific CD8+ T-cell reactivity and high protection against B16 melanoma challenge. Unexpectedly, administration of LVs containing the MHCII promoter resulted in persistence of luciferase expression and viral integration in MHCII+ splenocytes and virtually no CD8+ T-cell responses against TRP2. These studies reveal that APC transduction by LVs could lead to immune reactivity (LV-CMV) or persistence of transgene expression (LV-MHCII), providing a relevant paradigm for vaccination and gene replacement approaches.
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U2 - 10.1038/sj.mt.6300180
DO - 10.1038/sj.mt.6300180
M3 - Article
C2 - 17505480
AN - SCOPUS:34250702619
SN - 1525-0016
VL - 15
SP - 1390
EP - 1399
JO - Molecular Therapy
JF - Molecular Therapy
IS - 7
ER -
