title: Lentiviral vectors with CMV or MHCII promoters administered in vivo: immune reactivity versus persistence of expression.
abstract: 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.
authors.affiliation: Department of Medicine, Division of Digestive Diseases, David Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, USA. rstripecke@mednet.ucla.edu
authors.forename: Takahiro
authors.initials: T
authors.lastname: Kimura
authors.forename: Richard C
authors.initials: RC
authors.lastname: Koya
authors.forename: Laura
authors.initials: L
authors.lastname: Anselmi
authors.forename: Catia
authors.initials: C
authors.lastname: Sternini
authors.forename: He-Jing
authors.initials: HJ
authors.lastname: Wang
authors.forename: Begonya
authors.initials: B
authors.lastname: Comin-Anduix
authors.forename: Robert M
authors.initials: RM
authors.lastname: Prins
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Faure-Kumar
authors.forename: Nora
authors.initials: N
authors.lastname: Rozengurt
authors.forename: Yan
authors.initials: Y
authors.lastname: Cui
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Kasahara
authors.forename: Renata
authors.initials: R
authors.lastname: Stripecke
mesh_terms: D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D045744:Cell Line, Tumor; D003587:Cytomegalovirus / Q000235:genetics*; D005260:Female; D015870:Gene Expression / Q000235:genetics*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics*; D000949:Histocompatibility Antigens Class II / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics*; D008099:Liver / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012680:Sensitivity and Specificity; D013154:Spleen / Q000276:immunology / Q000378:metabolism; D015996:Survival Rate; D019076:Transgenes / Q000235:genetics; D014611:Vaccination; D053586:Virus Internalization
chemical_list: D000949:Histocompatibility Antigens Class II