Dendritic cell–targeted lentiviral vector immunization uses pseudotransduction and DNA-mediated STING and cGAS activation
Abstract
Dendritic cell (DC) activation and antigen presentation are critical for efficient priming of T cell responses. Here, we study how lentiviral vectors (LVs) deliver antigen and activate DCs to generate T cell immunization in vivo. We report that antigenic proteins delivered in vector particles via pseudotransduction were sufficient to stimulate an antigen-specific immune response. The delivery of the viral genome encoding the antigen increased the magnitude of this response in vivo but was irrelevant in vitro. Activation of DCs by LVs was independent of MyD88, TRIF, and MAVS, ruling out an involvement of Toll-like receptor or RIG-I–like receptor signaling. Cellular DNA packaged in LV preparations induced DC activation by the host STING (stimulator of interferon genes) and cGAS (cyclic guanosine monophosphate–adenosine monophosphate synthase) pathway. Envelope-mediated viral fusion also activated DCs in a phosphoinositide 3-kinase–dependent but STING-independent process. Pseudotransduction, transduction, viral fusion, and delivery of cellular DNA collaborate to make the DC-targeted LV preparation an effective immunogen.
Additional Information
© 2017 American Association for the Advancement of Science. Submitted 30 September 2016; Resubmitted 14 March 2017; Accepted 12 June 2017; Published 21 July 2017. We thank G. Cheng (UCLA) for the MAVS−/− mice, Z. Chen (University of Texas Southwestern Medical Center) for the cGAS−/− mice, the UCLA/CFAR Virology Core Laboratory (5P30 AI028697), the UCLA Specialty Training and Advanced Research Program, the AIDS Reagent Program, the National Institute of Allergy and Infectious Diseases Tetramer Core Facility, the UCLA/CFAR Virology Core Laboratory, and the PrimerBank. This work was supported by the Millard and Muriel Jacobs Genetics and Genomics Laboratory at Caltech. We thank M. Bethune and G. Li for the review of the manuscript. Funding: This work was supported by the NIH (OPPGH5157) and the American Foundation for AIDS Research (108292-51-RGRL). J.T.K. was supported by the NIH (KL2TR001882) and the UCLA Specialty Training and Advanced Research (STAR) Program. Author contributions: J.T.K. designed the research, performed the experiments, analyzed the results, and drafted the manuscript. Y.L., B.D., G.L., R.P.K., K.K.L., and Y.O. performed the experiments. L.Y. and P.W. discussed the results and drafted the manuscript. D.B. designed the research, discussed the results, drafted the manuscript, and provided financial support. Competing interests: P.W., L.Y., and D.B. are inventors on issued patents for DC-targeted lentiviral technology discussed in this study, which has been licensed to Immune Design Corporation (IDC), and are stock holders in IDC. D.B. is also a member of Board of Directors of IDC. Data and materials availability: DNA sequence data have been deposited in Sequence Read Archive under accession number SAMN07195639.Attached Files
Accepted Version - nihms933217.pdf
Supplemental Material - aal1329_SM.pdf
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Additional details
- PMCID
- PMC5845465
- Eprint ID
- 79147
- DOI
- 10.1126/sciimmunol.aal1329
- Resolver ID
- CaltechAUTHORS:20170718-091900989
- NIH
- OPPGH5157
- American Foundation for AIDS Research
- 108292-51-RGRL
- NIH
- KL2TR001882
- UCLA Specialty Training and Advanced Research (STAR) Program
- Millard and Muriel Jacobs Genetics and Genomics Laboratory
- Created
-
2017-07-21Created from EPrint's datestamp field
- Updated
-
2022-03-23Created from EPrint's last_modified field
- Caltech groups
- Millard and Muriel Jacobs Genetics and Genomics Laboratory