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Single-round infectious particles enhance immunogenicity of a DNA vaccine against West Nile virus

Nature Biotechnology volume 26, pages 571–577 (2008)Cite this article

Abstract

DNA vaccines encoding replication-defective viruses are safer than inactivated or live attenuated viruses but may fail to stimulate an immune response sufficient for effective vaccination. We augment the protective capacity of a capsid-deleted flavivirus DNA vaccine by co-expressing the capsid protein from a separate promoter. In transfected cells, the capsid-deleted RNA transcript is replicated and translated to produce secreted virus-like particles lacking the nucleocapsid. This RNA is also packaged with the help of co-expressed capsid protein to form secreted single-round infectious particles (SRIPs) that deliver the RNA into neighboring cells. In SRIP-infected cells, the RNA is replicated again and produces additional virus-like particles, but in the absence of capsid RNA no SRIPs are formed and no further spread occurs. Compared with an otherwise identical construct that does not encode capsid, our vaccine offers better protection to mice after lethal West Nile virus infection. It also elicits virus-neutralizing antibodies in horses. This approach may enable vaccination against pathogenic flaviviruses other than West Nile virus.

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Figure 1: Structure and mode of operation of a split genome–based DNA vaccine against a flavivirus.
Figure 2: Production of SRIPs from pKUNdC/C DNA in vitro and ex vivo.
Figure 3: Immunogenicity of pKUNdC/C DNA in mice and horses.

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Acknowledgements

We thank Jane Pollitt, Joanne Meers and Paul Mills (all from School of Veterinary Sciences, University of Queensland) for valuable assistance in designing and performing horse vaccination experiments; Wai Yuen Cheah, Kim Pham and Natalie Prow (University of Queensland) for assistance with mouse experiments and serological analysis of mouse and horse sera; Greg Smith, Alyssa Pyke and Bruce Harrower (all from Queensland Heath Forensic and Scientific Services) for generously providing PC3 animal facilities and logistical assistance for mouse challenge studies; and James Doecke (QIMR) for assistance with statistics.

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Authors and Affiliations

  1. School of Molecular and Microbial Sciences, University of Queensland, St. Lucia, Brisbane, 4072, Queensland, Australia

    David C Chang, Wen J Liu, David C Clark, Roy A Hall & Alexander A Khromykh

  2. Queensland Institute of Medical Research, PO Royal Brisbane Hospital, Brisbane, 4029, Queensland, Australia

    Itaru Anraku & Andreas Suhrbier

  3. School of Veterinary Sciences, University of Queensland, St. Lucia, Brisbane, 4072, Queensland, Australia

    Christopher C Pollitt

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Contributions

D.C. Chang designed experiments, performed the majority of the experiments, analyzed data and wrote the manuscript; W.J.L. designed experiments, immunized mice and analyzed data; I.A. designed, performed and analyzed ELISPOT assays; D.C. Clark assisted with mice experiments; C.C.P. provided veterinary expertise and supervised horse experiments; A.S. designed the ELISPOT assays and wrote the manuscript; R.A.H. designed animal experiments, analyzed data and wrote the manuscript; and A.A.K. generated the idea, designed and coordinated the research, analyzed data and wrote the manuscript.

Corresponding author

Correspondence to Alexander A Khromykh.

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Chang, D., Liu, W., Anraku, I. et al. Single-round infectious particles enhance immunogenicity of a DNA vaccine against West Nile virus. Nat Biotechnol 26, 571–577 (2008). https://doi.org/10.1038/nbt1400

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