Letter | Published:

Hexon-chimaeric adenovirus serotype 5 vectors circumvent pre-existing anti-vector immunity

Abstract

A common viral immune evasion strategy involves mutating viral surface proteins in order to evade host neutralizing antibodies. Such immune evasion tactics have not previously been intentionally applied to the development of novel viral gene delivery vectors that overcome the critical problem of anti-vector immunity. Recombinant, replication-incompetent adenovirus serotype 5 (rAd5) vector-based vaccines for human immunodeficiency virus type 1 and other pathogens have proved highly immunogenic in preclinical studies1,2 but will probably be limited by the high prevalence of pre-existing anti-Ad5 immunity in human populations, particularly in the developing world3,4,5,6. Here we show that rAd5 vectors can be engineered to circumvent anti-Ad5 immunity. We constructed novel chimaeric rAd5 vectors in which the seven short hypervariable regions (HVRs) on the surface of the Ad5 hexon protein were replaced with the corresponding HVRs from the rare adenovirus serotype Ad48. These HVR-chimaeric rAd5 vectors were produced at high titres and were stable through serial passages in vitro. HVR-chimaeric rAd5 vectors expressing simian immunodeficiency virus Gag proved comparably immunogenic to parental rAd5 vectors in naive mice and rhesus monkeys. In the presence of high levels of pre-existing anti-Ad5 immunity, the immunogenicity of HVR-chimaeric rAd5 vectors was not detectably suppressed, whereas the immunogenicity of parental rAd5 vectors was abrogated. These data demonstrate that functionally relevant Ad5-specific neutralizing antibodies are focused on epitopes located within the hexon HVRs. Moreover, these studies show that recombinant viral vectors can be engineered to circumvent pre-existing anti-vector immunity by removing key neutralizing epitopes on the surface of viral capsid proteins. Such chimaeric viral vectors may have important practical implications for vaccination and gene therapy.

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References

  1. 1

    Shiver, J. W. et al. Replication-incompetent adenoviral vaccine vector elicits effective anti-immunodeficiency-virus immunity. Nature 415, 331–335 (2002)

  2. 2

    Sullivan, N. J. et al. Accelerated vaccination for Ebola virus haemorrhagic fever in non-human primates. Nature 424, 681–684 (2003)

  3. 3

    Shiver, J. W. & Emini, E. A. Recent advances in the development of HIV-1 vaccines using replication-incompetent adenovirus vectors. Annu. Rev. Med. 55, 355–372 (2004)

  4. 4

    Vogels, R. et al. Replication-deficient human adenovirus type 35 vectors for gene transfer and vaccination: efficient human cell interaction and bypass of preexisting adenovirus immunity. J. Virol. 77, 8263–8271 (2003)

  5. 5

    Kostense, S. et al. Adenovirus types 5 and 35 seroprevalence in AIDS risk groups supports type 35 as a vaccine vector. AIDS 18, 1213–1216 (2004)

  6. 6

    Sumida, S. M. et al. Neutralizing antibodies to adenovirus serotype 5 vaccine vectors are directed primarily against the adenovirus hexon protein. J. Immunol. 174, 7179–7185 (2005)

  7. 7

    Barouch, D. H. et al. Immunogenicity of recombinant adenovirus serotype 35 vaccine in the presence of pre-existing anti-Ad5 immunity. J. Immunol. 172, 6290–6297 (2004)

  8. 8

    Casimiro, D. R. et al. Comparative immunogenicity in rhesus monkeys of DNA plasmid, recombinant vaccinia virus, and replication-defective adenovirus vectors expressing a human immunodeficiency virus type 1 gag gene. J. Virol. 77, 6305–6313 (2003)

  9. 9

    Youil, R. et al. Hexon gene switch strategy for the generation of chimeric recombinant adenovirus. Hum. Gene Ther. 13, 311–320 (2002)

  10. 10

    Gall, J. G., Crystal, R. G. & Falck-Pedersen, E. Construction and characterization of hexon-chimeric adenoviruses: specification of adenovirus serotype. J. Virol. 72, 10260–10264 (1998)

  11. 11

    Crawford-Miksza, L. & Schnurr, D. P. Analysis of 15 adenovirus hexon proteins reveals the location and structure of seven hypervariable regions containing serotype-specific residues. J. Virol. 70, 1836–1844 (1996)

  12. 12

    Athappilly, F. K., Murali, R., Rux, J. J., Cai, Z. & Burnett, R. M. The refined crystal structure of hexon, the major coat protein of adenovirus type 2, at 2.9 Å resolution. J. Mol. Biol. 242, 430–455 (1994)

  13. 13

    Rux, J. J., Kuser, P. R. & Burnett, R. M. Structural and phylogenetic analysis of adenovirus hexons by use of high-resolution x-ray crystallographic, molecular modeling, and sequence-based methods. J. Virol. 77, 9553–9566 (2003)

  14. 14

    Jones, T. A., Zou, J. Y., Cowan, S. W. & Kjeldgaard, M. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr. A 47, 110–119 (1991)

  15. 15

    Kraulis, P. J. MOLSCRIPT: A program to produce both detailed and schematic plots of protein structures. J. Appl. Crystallogr. 24, 946–950 (1991)

  16. 16

    Sprangers, M. C. et al. Quantifying adenovirus-neutralizing antibodies by luciferase transgene detection: addressing preexisting immunity to vaccine and gene therapy vectors. J. Clin. Microbiol. 41, 5046–5052 (2003)

  17. 17

    Pitcher, C. J. et al. Development and homeostasis of T cell memory in rhesus macaque. J. Immunol. 168, 29–43 (2002)

  18. 18

    Gahery-Segard, H. et al. Immune response to recombinant capsid proteins of adenovirus in humans: antifiber and anti-penton base antibodies have a synergistic effect on neutralizing activity. J. Virol. 72, 2388–2397 (1998)

  19. 19

    Hong, S. S., Habib, N. A., Franqueville, L., Jensen, S. & Boulanger, P. A. Identification of adenovirus (Ad) penton base neutralizing epitopes by use of sera from patients who had received conditionally replicative Ad (Addl1520) for treatment of liver tumors. J. Virol. 77, 10366–10375 (2003)

  20. 20

    Wohlfart, C. Neutralization of adenoviruses: kinetics, stoichiometry, and mechanisms. J. Virol. 62, 2321–2328 (1988)

  21. 21

    Havenga, M. J. et al. Improved adenovirus vectors for infection of cardiovascular tissues. J. Virol. 75, 3335–3342 (2001)

  22. 22

    Nanda, A. et al. Immunogenicity of recombinant fiber-chimeric adenovirus serotype 35 vector-based vaccines in mice and rhesus monkeys. J. Virol. 79, 14161–14168 (2005)

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Acknowledgements

We thank R. Vogels, J. Custers, N. Letvin, R. Dolin, A. Kelcz, Y. Sun, L. Shen, M. Kishko, D. Truitt, S. Harrison, B. Walker and P. Kiepiela for advice, assistance and reagents. The peptide pools were obtained from the NIH AIDS Research and Reference Reagent Program. We acknowledge support from NIH grants to D.H.B.

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Correspondence to Dan H. Barouch.

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Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

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Further reading

Figure 1: Construction of hexon HVR-chimaeric rAd5 vectors.
Figure 2: Cellular immune responses elicited by hexon HVR-chimaeric rAd5 vectors.
Figure 3: Immunogenicity of heterologous recombinant adenovirus prime–boost regimens.
Figure 4: Humoral immune responses to hexon HVR-chimaeric rAd5 vectors.
Figure 5: Immunogenicity of hexon HVR-chimaeric rAd5 vectors in rhesus monkeys.

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