Technical Report | Published:

Development of replication-defective lymphocytic choriomeningitis virus vectors for the induction of potent CD8+ T cell immunity

Nature Medicine volume 16, pages 339345 (2010) | Download Citation

Abstract

Lymphocytic choriomeningitis virus (LCMV) exhibits natural tropism for dendritic cells and represents the prototypic infection that elicits protective CD8+ T cell (cytotoxic T lymphocyte (CTL)) immunity. Here we have harnessed the immunobiology of this arenavirus for vaccine delivery. By using producer cells constitutively synthesizing the viral glycoprotein (GP), it was possible to replace the gene encoding LCMV GP with vaccine antigens to create replication-defective vaccine vectors. These rLCMV vaccines elicited CTL responses that were equivalent to or greater than those elicited by recombinant adenovirus 5 or recombinant vaccinia virus in their magnitude and cytokine profiles, and they exhibited more effective protection in several models. In contrast to recombinant adenovirus 5, rLCMV failed to elicit vector-specific antibody immunity, which facilitated re-administration of the same vector for booster vaccination. In addition, rLCMV elicited T helper type 1 CD4+ T cell responses and protective neutralizing antibodies to vaccine antigens. These features, together with low seroprevalence in humans, suggest that rLCMV may show utility as a vaccine platform against infectious diseases and cancer.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from $8.99

All prices are NET prices.

References

  1. 1.

    et al. An HIV-1 clade C DNA prime, NYVAC boost vaccine regimen induces reliable, polyfunctional, and long-lasting T cell responses. J. Exp. Med. 205, 63–77 (2008).

  2. 2.

    et al. Studies of a prophylactic HIV-1 vaccine candidate based on modified vaccinia virus Ankara (MVA) with and without DNA priming: effects of dosage and route on safety and immunogenicity. Vaccine 25, 2120–2127 (2007).

  3. 3.

    et al. Multifunctional TH1 cells define a correlate of vaccine-mediated protection against Leishmania major. Nat. Med. 13, 843–850 (2007).

  4. 4.

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

  5. 5.

    et al. Hexon-chimaeric adenovirus serotype 5 vectors circumvent pre-existing anti-vector immunity. Nature 441, 239–243 (2006).

  6. 6.

    , & Arenaviridae: The viruses and their replication. in Fields Virology (eds. Knipe, D.M. and Howley, P.M.) 1635–1668 (Lippincott Williams & Wilkins, Philadelphia, Pennsylvania, 2001).

  7. 7.

    , & Differential regulation of antiviral T-cell immunity results in stable CD8+ but declining CD4+ T-cell memory. Nat. Med. 7, 913–919 (2001).

  8. 8.

    et al. Kinetics of protective antibodies are determined by the viral surface antigen. J. Clin. Invest. 114, 988–993 (2004).

  9. 9.

    , , & Recombinant lymphocytic choriomeningitis virus expressing vesicular stomatitis virus glycoprotein. Proc. Natl. Acad. Sci. USA 100, 7895–7900 (2003).

  10. 10.

    , , & Recovery of an arenavirus entirely from RNA polymerase I/II-driven cDNA. Proc. Natl. Acad. Sci. USA 103, 4663–4668 (2006).

  11. 11.

    et al. Persistence of lymphocytic choriomeningitis virus at very low levels in immune mice. Proc. Natl. Acad. Sci. USA 96, 11964–11969 (1999).

  12. 12.

    , , & Rapid functional exhaustion and deletion of CTL following immunization with recombinant adenovirus. J. Immunol. 174, 4559–4566 (2005).

  13. 13.

    et al. Long-lived virus-reactive memory T cells generated from purified cytokine-secreting T helper type 1 and type 2 effectors. J. Exp. Med. 205, 53–61 (2008).

  14. 14.

    et al. Antiviral protection by vesicular stomatitis virus-specific antibodies in alpha/beta interferon receptor-deficient mice. J. Virol. 69, 2153–2158 (1995).

  15. 15.

    et al. Organ-specific regulation of the CD8 T cell response to Listeria monocytogenes infection. J. Immunol. 166, 3402–3409 (2001).

  16. 16.

    , , , & Recombinant Listeria monocytogenes as a live vaccine vehicle and a probe for studying cell-mediated immunity. Immunol. Rev. 158, 147–157 (1997).

  17. 17.

    et al. CD8 T cell ignorance or tolerance to islet antigens depends on antigen dose. Proc. Natl. Acad. Sci. USA 96, 12703–12707 (1999).

  18. 18.

    et al. T cell receptor antagonist peptides induce positive selection. Cell 76, 17–27 (1994).

  19. 19.

    et al. Tumor-infiltrating lymphocytes exhibiting high ex vivo cytolytic activity fail to prevent murine melanoma tumor growth in vivo. J. Immunol. 161, 2187–2194 (1998).

  20. 20.

    Dendritic cells: versatile controllers of the immune system. Nat. Med. 13, 1155–1159 (2007).

  21. 21.

    , , & Inducible transgenic mice reveal resting dendritic cells as potent inducers of CD8+ T cell tolerance. Immunity 18, 713–720 (2003).

  22. 22.

    et al. Immunosuppression and resultant viral persistence by specific viral targeting of dendritic cells. J. Exp. Med. 192, 1249–1260 (2000).

  23. 23.

    , , , & Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon Cre-mediated excision. Genesis 28, 147–155 (2000).

  24. 24.

    et al. Induction of protective cytotoxic T cell responses in the presence of high titers of virus-neutralizing antibodies: implications for passive and active immunization. J. Exp. Med. 187, 649–654 (1998).

  25. 25.

    , & Prevalence of antibodies to lymphocytic choriomeningitis virus in blood donors in southeastern France. Transfusion 47, 172–173 (2007).

  26. 26.

    , , , & Lymphocytic choriomeningitis virus infection in a province of Spain: analysis of sera from the general population and wild rodents. J. Med. Virol. 70, 273–275 (2003).

  27. 27.

    et al. Low prevalence of antibodies against the zoonotic agents Brucella abortus, Leptospira spp., Streptococcus suis serotype II, hantavirus, and lymphocytic choriomeningitis virus among veterinarians and pig farmers in the southern part of The Netherlands. Vet. Q. 21, 50–54 (1999).

  28. 28.

    et al. Prevalence of serum antibodies against lymphocytic choriomeningitis virus in selected populations from two U.S. cities. J. Med. Virol. 38, 27–31 (1992).

  29. 29.

    et al. Transmission of lymphocytic choriomeningitis virus by organ transplantation. N. Engl. J. Med. 354, 2235–2249 (2006).

  30. 30.

    et al. Multiclade human immunodeficiency virus type 1 envelope immunogens elicit broad cellular and humoral immunity in rhesus monkeys. J. Virol. 79, 2956–2963 (2005).

  31. 31.

    et al. Decreased tumor surveillance after adoptive T-cell therapy. Cancer Res. 67, 7467–7476 (2007).

  32. 32.

    et al. Induction of potent antitumor CTL responses by recombinant vaccinia encoding a melan-A peptide analogue. J. Immunol. 164, 1125–1131 (2000).

  33. 33.

    et al. DNA vaccines encoding retrovirus-based virus-like particles induce efficient immune responses without adjuvant. Vaccine 24, 2643–2655 (2006).

  34. 34.

    et al. Fully detargeted polyethylene glycol-coated adenovirus vectors are potent genetic vaccines and escape from pre-existing anti-adenovirus antibodies. Mol. Ther. 16, 154–162 (2008).

  35. 35.

    et al. Dissociation of proteasomal degradation of biosynthesized viral proteins from generation of MHC class I-associated antigenic peptides. J. Immunol. 160, 4859–4868 (1998).

  36. 36.

    et al. Anti-viral protection and prevention of lymphocytic choriomeningitis or of the local footpad swelling reaction in mice by immunization with vaccinia-recombinant virus expressing LCMV-WE nucleoprotein or glycoprotein. Eur. J. Immunol. 19, 417–424 (1989).

  37. 37.

    et al. Immunodominance of an antiviral cytotoxic T cell response is shaped by the kinetics of viral protein expression. J. Immunol. 171, 5415–5422 (2003).

  38. 38.

    , , , & Tolerance induction in double specific T-cell receptor transgenic mice varies with antigen. Nature 342, 559–561 (1989).

  39. 39.

    et al. HLA-A2.1-restricted education and cytolytic activity of CD8+ T lymphocytes from β2 microglobulin (β2m) HLA-A2.1 monochain transgenic H-2Db β2m double knockout mice. J. Exp. Med. 185, 2043–2051 (1997).

  40. 40.

    et al. Toll-like receptor ligands modulate dendritic cells to augment cytomegalovirus- and HIV-1-specific T cell responses. J. Immunol. 171, 4320–4328 (2003).

Download references

Acknowledgements

We thank H. Hengartner and R. Zinkernagel for critical comments, suggestions, discussions and long-term support; E. Horvath for technical assistance; S.A. Rosenberg and J.R. Wunderlich for samples from patients with melanoma; M. Roederer and K. Foulds for reagents and flow cytometry support; A. Oxenius, R. Spörri and N. Joller for providing access to their flow cytometry facility; A. Pegu, R. Roychoudhuri and C. Cheng for discussions and advice on human DC cultures; D. von Laer (Georg-Speyer-Haus) for plasmid M369 and GP-expressing 293T cells; H. Shen (University of Pennsylvania School of Medicine) for rLM-OVA; M. Groettrup (University of Constance) for VACC-OVA, originally generated by J. Yewdell (National Institute of Allergy and Infectious Diseases); and R. Schirmbeck (University of Ulm) for StT-OVA-G cDNA. L.F. was supported by a fellowship of the Schweizerische Stiftung für medizinisch-biologische Stipendien. A.N.H. is a fellow of GRAKO1121 of the German Research Foundation. M.L. is a Lichtenberg fellow funded by the Volkswagen Foundation. A.B. was supported by a PhD scholarship of the Boehringer Ingelheim Fonds and by a post-doctoral fellowship of the Roche Research Foundation. D.D.P. holds a stipendiary professorship of the Swiss National Science Foundation (PP00A-114913) and was supported by grant 3100A0-104067/1 of the Swiss National Science Foundation.

Author information

Author notes

    • Frédéric Lévy

    Present address: Debiopharm, Lausanne, Switzerland.

    • Ahmed N Hegazy
    • , Andreas Bergthaler
    •  & Admar Verschoor

    These authors contributed equally to this work.

Affiliations

  1. Department of Pathology and Immunology, University of Geneva, Geneva, Switzerland.

    • Lukas Flatz
    • , Andreas Bergthaler
    • , Marylise Fernandez
    • , Susan Johnson
    • , Claire-Anne Siegrist
    •  & Daniel D Pinschewer
  2. Institute of Experimental Immunology, University Hospital of Zurich, Zurich, Switzerland.

    • Lukas Flatz
    • , Ahmed N Hegazy
    • , Andreas Bergthaler
    • , Admar Verschoor
    • , Maries van den Broek
    • , Max Löhning
    •  & Daniel D Pinschewer
  3. Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.

    • Lukas Flatz
    •  & Gary J Nabel
  4. Experimental Immunology, Department of Rheumatology and Clinical Immunology, Charité–University Medicine, Berlin, Germany.

    • Ahmed N Hegazy
    • , Andreas Radbruch
    •  & Max Löhning
  5. Deutsches Rheuma-Forschungszentrum, Berlin, Germany.

    • Ahmed N Hegazy
    • , Andreas Radbruch
    •  & Max Löhning
  6. Institute for Systems Biology, Seattle, Washington, USA.

    • Andreas Bergthaler
  7. Institute for Medical Microbiology, Immunology and Hygiene, Technical University Munich, Munich, Germany.

    • Admar Verschoor
  8. Tumor Immunology, Department of Clinical Research, University of Berne, Berne, Switzerland.

    • Christina Claus
    •  & Adrian F Ochsenbein
  9. World Health Organization Collaborating Center for Neonatal Vaccinology, University of Geneva, Geneva, Switzerland.

    • Marylise Fernandez
    • , Susan Johnson
    • , Paul-Henri Lambert
    • , Claire-Anne Siegrist
    •  & Daniel D Pinschewer
  10. Center for Cancer Research, National Cancer Institute, US National Institutes of Health, Bethesda, Maryland, USA.

    • Luca Gattinoni
    •  & Nicholas P Restifo
  11. Division of Gene Therapy, University of Ulm, Ulm, Germany.

    • Florian Kreppel
    •  & Stefan Kochanek
  12. Oncology, University Hospital of Zurich, Zurich, Switzerland.

    • Maries van den Broek
  13. Ludwig Institute for Cancer Research, Epalinges, Switzerland.

    • Frédéric Lévy
  14. Department of Pediatrics, University of Geneva, Geneva, Switzerland.

    • Claire-Anne Siegrist

Authors

  1. Search for Lukas Flatz in:

  2. Search for Ahmed N Hegazy in:

  3. Search for Andreas Bergthaler in:

  4. Search for Admar Verschoor in:

  5. Search for Christina Claus in:

  6. Search for Marylise Fernandez in:

  7. Search for Luca Gattinoni in:

  8. Search for Susan Johnson in:

  9. Search for Florian Kreppel in:

  10. Search for Stefan Kochanek in:

  11. Search for Maries van den Broek in:

  12. Search for Andreas Radbruch in:

  13. Search for Frédéric Lévy in:

  14. Search for Paul-Henri Lambert in:

  15. Search for Claire-Anne Siegrist in:

  16. Search for Nicholas P Restifo in:

  17. Search for Max Löhning in:

  18. Search for Adrian F Ochsenbein in:

  19. Search for Gary J Nabel in:

  20. Search for Daniel D Pinschewer in:

Contributions

L.F., A.N.H., A.B., A.V., C.C., M.F., L.G., S.J., F.K. and D.D.P. performed experiments; L.F., A.N.H., A.B., A.V., C.C., L.G., P.-H.L., C.-A.S., N.P.R., M.L., A.F.O., G.J.N. and D.D.P. designed experiments; S.K., M.v.d.B., A.R. and F.L. contributed reagents; and L.F., G.J.N. and D.D.P. wrote the manuscript.

Competing interests

L.F., A.B. and D.D.P. are listed as co-inventors on a patent held by the University of Zurich on arenavirus vectors and thus they will be recipients of potential future revenues from this intellectual property. C.A.S. has received honoraria for participation in scientific advisory boards and research grants from several vaccine manufacturers, none related to this work.

Corresponding authors

Correspondence to Lukas Flatz or Daniel D Pinschewer.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–8

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nm.2104

COMPETING INTERESTS STATEMENT

The authors declare competing financial interests: details accompany the full-text HTML version of the paper at http://www.nature.com/naturemedicine/.