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: Takahiro Kimura, Richard C Koya, Laura Anselmi, Catia Sternini, He-Jing Wang, Begonya Comin-Anduix, Robert M Prins, Emmanuelle Faure-Kumar, Nora Rozengurt, Yan Cui, Noriyuki Kasahara, Renata Stripecke
• Journal: Molecular therapy : the journal of the American Society of Gene Therapy (Mol Ther) Vol. 15 Issue 7 Pg. 1390-9 (Jul 2007) ISSN: 1525-0024 [Electronic] United States
• PMID: 17505480 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Histocompatibility Antigens Class II

Topics

• Animals
• CD8-Positive T-Lymphocytes (immunology, metabolism)
• Cell Line, Tumor
• Cytomegalovirus (genetics)
• Female
• Gene Expression (genetics)
• Genetic Vectors (administration & dosage, genetics)
• Histocompatibility Antigens Class II (genetics, immunology, metabolism)
• Lentivirus (genetics)
• Liver (metabolism)
• Mice
• Mice, Inbred C57BL
• Mice, Nude
• Neoplasm Transplantation
• Promoter Regions, Genetic (genetics)
• Sensitivity and Specificity
• Spleen (immunology, metabolism)
• Survival Rate
• Transgenes (genetics)
• Vaccination
• Virus Internalization