Vaccine protection against Zika virus from Brazil

Journal name:
Nature
Volume:
536,
Pages:
474–478
Date published:
DOI:
doi:10.1038/nature18952
Received
Accepted
Published online

Zika virus (ZIKV) is a flavivirus that is responsible for the current epidemic in Brazil and the Americas1, 2. ZIKV has been causally associated with fetal microcephaly, intrauterine growth restriction, and other birth defects in both humans3, 4, 5, 6, 7, 8 and mice9, 10, 11. The rapid development of a safe and effective ZIKV vaccine is a global health priority1, 2, but very little is currently known about ZIKV immunology and mechanisms of immune protection. Here we show that a single immunization with a plasmid DNA vaccine or a purified inactivated virus vaccine provides complete protection in susceptible mice against challenge with a strain of ZIKV involved in the outbreak in northeast Brazil. This ZIKV strain has recently been shown to cross the placenta and to induce fetal microcephaly and other congenital malformations in mice11. We produced DNA vaccines expressing ZIKV pre-membrane and envelope (prM-Env), as well as a series of deletion mutants. The prM-Env DNA vaccine, but not the deletion mutants, afforded complete protection against ZIKV, as measured by absence of detectable viraemia following challenge, and protective efficacy correlated with Env-specific antibody titers. Adoptive transfer of purified IgG from vaccinated mice conferred passive protection, and depletion of CD4 and CD8 T lymphocytes in vaccinated mice did not abrogate this protection. These data demonstrate that protection against ZIKV challenge can be achieved by single-shot subunit and inactivated virus vaccines in mice and that Env-specific antibody titers represent key immunologic correlates of protection. Our findings suggest that the development of a ZIKV vaccine for humans is likely to be achievable.

At a glance

Figures

  1. Construction and immunogenicity of DNA vaccines.
    Figure 1: Construction and immunogenicity of DNA vaccines.

    a, Schema of ZIKV prM-Env immunogens and deletion mutants. b, Western blot of transgene expression from (1) prM-Env, (2) prM-Env(ΔTM), (3) prM-Env(Δstem), (4) Env, (5) Env(ΔTM), (6) Env(Δstem), and (7) sham DNA vaccines transfected in 293T cells. Balb/c mice (n = 5 per group) received a single immunization with 50 μg of these DNA vaccines by the i.m. route. c, Humoral immune responses were assessed at week 3 following vaccination by Env-specific ELISA. Red bars reflect medians. d, e, Cellular immune responses were assessed by IFNγ ELISPOT assays (d) and multi-parameter intracellular cytokine staining assays (e). Error bars reflect s.e.m.

  2. Protective efficacy of DNA vaccines.
    Figure 2: Protective efficacy of DNA vaccines.

    a, Balb/c mice (n = 5 or 10 per group) received a single immunization by the i.m. route with 50 μg prM-Env DNA vaccine or a sham vaccine and were challenged at week 4 by the i.v. route with 105 viral particles (102 PFU) ZIKV-BR or ZIKV-PR. Serum viral loads are shown. b, Mice (n = 5 per group) received a single immunization with 50 μg of various DNA vaccines and were challenged with ZIKV-BR. c, d, Correlates of protective efficacy (c) and day 3 viral loads (d) are shown. Red bars reflect medians. P values and R2 values reflect t-tests and Spearman rank-correlation tests.

  3. Mechanistic studies.
    Figure 3: Mechanistic studies.

    a, Env-specific serum antibody titers in recipient Balb/c mice (n = 5 per group) following adoptive transfer of varying amounts (high, mid, low) of IgG purified from serum from mice vaccinated with prM-Env DNA or naive mice (sham). b, Correlates of protective efficacy. c, Serum viral loads in mice that received adoptive transfer of purified IgG from vaccinated mice and were challenged with ZIKV-BR. d, Serum viral loads in prM-Env-DNA-vaccinated mice that were depleted of CD4+ and/or CD8+ T lymphocytes before challenge with ZIKV-BR. Red bars reflect medians. P values reflect t-tests.

  4. Immunogenicity and protective efficacy of the PIV vaccine.
    Figure 4: Immunogenicity and protective efficacy of the PIV vaccine.

    Balb/c mice (n = 5 per group) received a single immunization by the i.m. or s.c. route with 1 μg PIV vaccine with alum, or alum alone, and were challenged at week 4 by the i.v. route with 105 viral particles (102 PFU) ZIKV-BR. a, Humoral immune responses were assessed at week 3 following vaccination by Env-specific ELISA. b, Correlates of protective efficacy. c, Serum viral loads are shown following ZIKV-BR challenge. Red bars reflect medians. P values reflect t-tests.

  5. ZIKV maximum likelihood phylogenetic tree.
    Extended Data Fig. 1: ZIKV maximum likelihood phylogenetic tree.

    The ZIKV-BR and ZIKV-PR challenge isolates are depicted with red arrows.

  6. ZIKV amino acid sequence comparisons.
    Extended Data Fig. 2: ZIKV amino acid sequence comparisons.

    Number of and percentage of amino acid differences in the polyprotein are shown for the following ZIKV isolates: Brazil/ZKV2015 (Brazil strain; ZIKV-BR challenge stock), PRVABC59 (Puerto Rico strain; ZIKV-PR challenge stock), BeH815744 (Brazil strain; immunogen design), H/PF/2013 (French Polynesian strain), and MR766 (African strain).

  7. prM-specific antibody responses in DNA-vaccinated mice.
    Extended Data Fig. 3: prM-specific antibody responses in DNA-vaccinated mice.

    In the experiment described in Fig. 2, humoral immune responses were assessed at week 3 following vaccination by prM-specific ELISA. Red bars reflect medians.

  8. Immunogenicity and protective efficacy of prM-Env DNA vaccine in SJL mice.
    Extended Data Fig. 4: Immunogenicity and protective efficacy of prM-Env DNA vaccine in SJL mice.

    SJL mice (n = 5 per group) received a single immunization by the i.m. route with 50 μg prM-Env DNA vaccine or a sham vaccine and were challenged at week 4 by the i.v. route with 105 viral particles (102 PFU) ZIKV-BR. Humoral immune responses were assessed at week 3 after vaccination by Env-specific ELISA (top). Red bars reflect medians. Serum viral loads are shown following ZIKV-BR challenge (bottom).

  9. Protective efficacy of prM-Env DNA vaccine in C57BL/6 mice.
    Extended Data Fig. 5: Protective efficacy of prM-Env DNA vaccine in C57BL/6 mice.

    C57BL/6 mice (n = 5 per group) received a single immunization by the i.m. route with 50 μg prM-Env DNA vaccine or a sham vaccine and were challenged at week 4 by the i.v. route with 105 viral particles (102 PFU) ZIKV-BR or ZIKV-PR. Serum viral loads are shown following challenge.

  10. Protective efficacy of various DNA vaccines in C57BL/6 mice.
    Extended Data Fig. 6: Protective efficacy of various DNA vaccines in C57BL/6 mice.

    C57BL/6 mice (n = 5 per group) received a single immunization by the i.m. route with 50 μg of various DNA vaccines and were challenged at week 4 by the i.v. route with 105 viral particles (102 PFU) ZIKV-BR. Serum viral loads are shown following challenge.

  11. Adoptive transfer of low titers of Env-specific IgG.
    Extended Data Fig. 7: Adoptive transfer of low titers of Env-specific IgG.

    Serum viral loads in mice that received adoptive transfer of low titers of Env-specific IgG (as defined in Fig. 3a) and were then challenged with ZIKV-BR.

  12. CD4+ and CD8+ T-lymphocyte depletion.
    Extended Data Fig. 8: CD4+ and CD8+ T-lymphocyte depletion.

    CD4+ and/or CD8+ T-lymphocyte depletion following monoclonal antibody treatment of Balb/c mice vaccinated with prM-Env DNA.

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Author information

  1. These authors contributed equally to this work.

    • Rafael A. Larocca &
    • Peter Abbink

Affiliations

  1. Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA

    • Rafael A. Larocca,
    • Peter Abbink,
    • M. Justin Iampietro,
    • Alexander Badamchi-Zadeh,
    • Michael Boyd,
    • David Ng’ang’a,
    • Marinela Kirilova,
    • Ramya Nityanandam,
    • Noe B. Mercado,
    • Zhenfeng Li,
    • Edward T. Moseley,
    • Christine A. Bricault,
    • Erica N. Borducchi,
    • Patricia B. Giglio,
    • David Jetton,
    • George Neubauer,
    • Joseph P. Nkolola,
    • Lori F. Maxfield &
    • Dan H. Barouch
  2. University of São Paulo, São Paulo 05508-000, Brazil

    • Jean Pierre S. Peron &
    • Paolo M. de A. Zanotto
  3. Walter Reed Army Institute of Research, Silver Spring, Maryland 20910, USA

    • Rafael A. De La Barrera,
    • Richard G. Jarman,
    • Kenneth H. Eckels,
    • Nelson L. Michael &
    • Stephen J. Thomas
  4. Ragon Institute of MGH, MIT, and Harvard, Cambridge, Massachusetts 02139, USA

    • Dan H. Barouch

Contributions

R.A.L., P.A. and D.H.B. designed the studies. J.P.S.P. and P.M.A.Z. developed the challenge virus. P.A., M.B., D.N., M.K., R.N., N.B.M. and Z.L. produced the DNA vaccines and conducted the virologic assays. R.A.B., R.G.J., K.H.E., N.L.M. and S.J.T. produced the PIV vaccines. R.A.L., M.J.I. and A.B.-Z. conducted the mouse studies. R.A.L., C.A.B., E.T.M., E.N.B., P.B.G., D.J., G.N., J.P.N., L.F.M., R.A.B. and R.G.J. conducted the immunologic assays. D.H.B. wrote the paper with all co-authors.

Competing financial interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to:

Reviewer Information

Nature thanks A. Barrett and G. Screaton for their contribution to the peer review of this work.

Author details

Extended data figures and tables

Extended Data Figures

  1. Extended Data Figure 1: ZIKV maximum likelihood phylogenetic tree. (839 KB)

    The ZIKV-BR and ZIKV-PR challenge isolates are depicted with red arrows.

  2. Extended Data Figure 2: ZIKV amino acid sequence comparisons. (189 KB)

    Number of and percentage of amino acid differences in the polyprotein are shown for the following ZIKV isolates: Brazil/ZKV2015 (Brazil strain; ZIKV-BR challenge stock), PRVABC59 (Puerto Rico strain; ZIKV-PR challenge stock), BeH815744 (Brazil strain; immunogen design), H/PF/2013 (French Polynesian strain), and MR766 (African strain).

  3. Extended Data Figure 3: prM-specific antibody responses in DNA-vaccinated mice. (168 KB)

    In the experiment described in Fig. 2, humoral immune responses were assessed at week 3 following vaccination by prM-specific ELISA. Red bars reflect medians.

  4. Extended Data Figure 4: Immunogenicity and protective efficacy of prM-Env DNA vaccine in SJL mice. (237 KB)

    SJL mice (n = 5 per group) received a single immunization by the i.m. route with 50 μg prM-Env DNA vaccine or a sham vaccine and were challenged at week 4 by the i.v. route with 105 viral particles (102 PFU) ZIKV-BR. Humoral immune responses were assessed at week 3 after vaccination by Env-specific ELISA (top). Red bars reflect medians. Serum viral loads are shown following ZIKV-BR challenge (bottom).

  5. Extended Data Figure 5: Protective efficacy of prM-Env DNA vaccine in C57BL/6 mice. (231 KB)

    C57BL/6 mice (n = 5 per group) received a single immunization by the i.m. route with 50 μg prM-Env DNA vaccine or a sham vaccine and were challenged at week 4 by the i.v. route with 105 viral particles (102 PFU) ZIKV-BR or ZIKV-PR. Serum viral loads are shown following challenge.

  6. Extended Data Figure 6: Protective efficacy of various DNA vaccines in C57BL/6 mice. (210 KB)

    C57BL/6 mice (n = 5 per group) received a single immunization by the i.m. route with 50 μg of various DNA vaccines and were challenged at week 4 by the i.v. route with 105 viral particles (102 PFU) ZIKV-BR. Serum viral loads are shown following challenge.

  7. Extended Data Figure 7: Adoptive transfer of low titers of Env-specific IgG. (166 KB)

    Serum viral loads in mice that received adoptive transfer of low titers of Env-specific IgG (as defined in Fig. 3a) and were then challenged with ZIKV-BR.

  8. Extended Data Figure 8: CD4+ and CD8+ T-lymphocyte depletion. (175 KB)

    CD4+ and/or CD8+ T-lymphocyte depletion following monoclonal antibody treatment of Balb/c mice vaccinated with prM-Env DNA.

Additional data