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A DNA-based vaccine protects against Crimean-Congo haemorrhagic fever virus disease in a Cynomolgus macaque model

Nature Microbiology volume 6, pages 187–195 (2021)Cite this article

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Abstract

There is currently no specific prophylaxis or vaccine against Crimean-Congo haemorrhagic fever virus (CCHFV). Crimean-Congo haemorrhagic fever (CCHF) is a severe febrile illness transmitted by Hyalomma ticks in endemic areas, handling of infected livestock or care of infected patients. We report here the successful protection against CCHFV-mediated disease in a non-human primate disease model. Cynomolgus macaques were vaccinated with a DNA-based vaccine using in vivo electroporation-assisted delivery. The vaccine contained two plasmids encoding the glycoprotein precursor (GPC) and the nucleoprotein (NP) of CCHFV. Animals received three vaccinations and we recorded potent antibody and T cell responses after vaccination. While all sham-vaccinated animals developed viraemia, high tissue viral loads and CCHF-induced disease, the NP + GPC vaccinated animals were significantly protected. In conclusion, this is evidence of a vaccine that can protect against CCHFV-induced disease in a non-human primate model. This supports clinical development of the vaccine to protect groups at risk for contracting the infection.

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Fig. 1: NP + GPC vaccination induces robust antibody responses to CCHFV.
Fig. 2: NP + GPC vaccination induces CCHFV-specific IFN-γ T cell responses.
Fig. 3: NP + GPC vaccination improves clinical scores and blood chemistry following CCHFV challenge.
Fig. 4: NP + GPC vaccination prevents viraemia and viral shedding.
Fig. 5: NP + GPC vaccination reduces viral burden in multiple tissues.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request and source data for Figs. 1 and 3–5 are provided. Source data are provided with this paper.

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Acknowledgements

We thank the staff of the Rocky Mountain Veterinary Branch for their help with animal care and veterinary clinical and pathology work. Our work was funded and supported by the CCHVaccine consortium that has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement no. 732732 (A.M., M.S. and F.W.), by the Swedish Research Council (Vetenskapsrådet, V.R. and M.S.) section Research Environment, Infection Biology (contract number 2018-05766) (A.M. and F.W.), the Swedish Cancer Foundation (M.S.), the Stockholm County Council ALF and CIMED grants (M.S.), Vinnova CAMP grant (M.S.) and by the Intramural Research Program of the NIAID, NIH. The funders had no role in study design, data interpretation or decision to publish.

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Author notes

  1. These authors contributed equally: Gustaf Ahlén, K. Sofia Appelberg, Heinz Feldmann, Matti Sällberg, Ali Mirazimi.

Authors and Affiliations

  1. Division of Intramural Research, NIAID/NIH, Hamilton, MT, USA

    David W. Hawman, Kimberly Meade-White, Patrick W. Hanley, Dana Scott, Atsushi Okumura & Heinz Feldmann

  2. Division of Clinical Microbiology, Department of Laboratory Medicine, Karolinska Institutet at Karolinska University Hospital Huddinge, Stockholm, Sweden

    Gustaf Ahlén, Vanessa Monteil, Matti Sällberg & Ali Mirazimi

  3. Public Health Agency of Sweden, Solna, Sweden

    K. Sofia Appelberg & Ali Mirazimi

  4. Institute for Virology, FB10-Veterinary Medicine, Justus Liebig University, Giessen, Germany

    Stephanie Devignot & Friedemann Weber

  5. National Veterinary Institute, Uppsala, Sweden

    Ali Mirazimi

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Contributions

D.W.H., H.F., G.A., M.S., F.W. and A.M. conceived and designed experiments. D.W.H., G.A., K.S.A., K.M.-W., P.W.H., D.S., V.M. and S.D. performed experiments. K.S.A. and G.A. contributed equally. D.W.H., P.W.H., D.S., A.O., M.S., H.F. and A.M. analysed data. D.W.H., H.F., M.S. and A.M. prepared the manuscript. All authors discussed the results and manuscript, and agreed to final publication. H.F., M.S. and A.M. contributed equally as senior authors.

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Correspondence to Ali Mirazimi.

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Competing interests

M.S. is a founder and chairman of the board of Svenska Vaccinfabriken Produktion AB.

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Extended data

Extended Data Fig. 1 Animal information and vaccination schedule.

(a) Animal number, date of birth (DOB), sex and vaccination. Animals were randomly assigned to either group prior to start of study. (b) Vaccination, blood draw and CCHFV challenge schedule. PV = post prime-vaccination, PI = post-CCHFV infection.

Extended Data Fig. 2 Vaccination Site Histology.

At Day +6 CCHFV challenge, 27 days after last vaccination, tissue from the vaccination site was fixed in 10% formalin and stained with hematoxylin and eosin. Slides were scored by a pathologist (a). 0 = absent, 1 = mild, 2 = moderate, 3 = severe. (b–d) Representative images of the one severe (b), one moderate (c) and one of six mild animals (d) are shown at 100x magnification.

Extended Data Fig. 3 CCHFV ELISA on serum from sham-vaccinated animals.

At indicated time points, serum was collected from sham-vaccinated animals and CCHFV-specific IgG measured by ELISA on day 0 or 56 postprime vaccination. N = 6 animals per timepoint. Data shown as mean plus standard deviation.

Extended Data Fig. 4 Sandwich ELISA.

A sandwich ELISA was performed on NP + GPC vaccinated animals to measure NP or Gc specific responses at day 0 post-prime vaccination (PV) and day 56 PV. N = 6 animals per timepoint. Statistical comparison performed using a two-way ANOVA with Sidak’s multiple comparison test.

Extended Data Fig. 5 Individual data for clinical scores and blood chemistry.

The data from Fig. 3 is shown again but with individual data points shown.

Extended Data Fig. 6 Anamnestic response to CCHFV challenge.

A whole-virion ELISA was used to measure antibody responses in animals after CCHFV challenge on day 6 post-infection (PI). CCHFV-specific IgG was quantified in a 1:6400 dilution of serum. Statistical tests performed using a two-way ANOVA with Sidak’s multiple comparison test.

Extended Data Fig. 7

General plasmid schematics of plasmids used in this study.

Supplementary information

Source data

Source Data Fig. 1

Statistical source data for Fig. 1 ELISA and neut data

Source Data Fig. 3

Statistical source data for Fig. 3

Source Data Fig. 4

Statistical source data for qPCR data shown in Fig. 4

Source Data Fig. 5

Statistical source data for qPCR data shown in Fig. 5

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Hawman, D.W., Ahlén, G., Appelberg, K.S. et al. A DNA-based vaccine protects against Crimean-Congo haemorrhagic fever virus disease in a Cynomolgus macaque model. Nat Microbiol 6, 187–195 (2021). https://doi.org/10.1038/s41564-020-00815-6

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