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mRNA vaccine against malaria tailored for liver-resident memory T cells

Nature Immunology volume 24, pages 1487–1498 (2023)Cite this article

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Abstract

Malaria is caused by Plasmodium species transmitted by Anopheles mosquitoes. Following a mosquito bite, Plasmodium sporozoites migrate from skin to liver, where extensive replication occurs, emerging later as merozoites that can infect red blood cells and cause symptoms of disease. As liver tissue-resident memory T cells (Trm cells) have recently been shown to control liver-stage infections, we embarked on a messenger RNA (mRNA)-based vaccine strategy to induce liver Trm cells to prevent malaria. Although a standard mRNA vaccine was unable to generate liver Trm or protect against challenge with Plasmodium berghei sporozoites in mice, addition of an agonist that recruits T cell help from type I natural killer T cells under mRNA-vaccination conditions resulted in significant generation of liver Trm cells and effective protection. Moreover, whereas previous exposure of mice to blood-stage infection impaired traditional vaccines based on attenuated sporozoites, mRNA vaccination was unaffected, underlining the potential for such a rational mRNA-based strategy in malaria-endemic regions.

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Fig. 1: Adjuvanted mRNA vaccines can induce liver Trm cells and protect against sporozoite challenge.
Fig. 2: Defining the ideal adjuvant for liver Trm cell induction.
Fig. 3: Dissection of the mechanistic basis for vaccine activity.
Fig. 4: Endogenous T cell response to adjuvanted mRNA vaccination.
Fig. 5: Vaccination with adjuvanted mRNA can induce protection against an authentic parasite antigen.
Fig. 6: Protective responses can be boosted and require CD8 cells.
Fig. 7: Protection is largely dependent on liver Trm cells.
Fig. 8: Vaccination with adjuvanted mRNA is not impaired by preexposure to blood-stage malaria.

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Data availability

Data to support the findings of study are available from the corresponding author upon request without restrictions. Source data are provided with this paper.

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Acknowledgements

We thank the BRF at the Peter Doherty Institute, and the BRU and the Hugh Green Cytometry Centre at the Malaghan Institute of Medical Research for technical support. We thank D. Bowen for support of K.E., D. Godfrey for the CD1d–PBS-44 tetramers and the NIH tetramer core facility for the CD1d–PBS-57 tetramers. This work was supported by funding from the New Zealand Ministry of Business Innovation and Employment (contract RTVU1603 to Victoria University of Wellington) and the New Zealand Health Research Council (contract HRC-20/569 to Victoria University of Wellington and HRC14/1003 Independent Research Organisation Fund to the Malaghan Institute). K.C.Y.P. is supported by the Monash Graduate Scholarship and the Monash University MNHS Faculty International Tuition Scholarship. D.F.R. was supported by the National Health and Medical Research Council of Australia (NHMRC) 1139486, S.G. by NHMRC 1159272, P.B. and K.E. by NHMRC 1146677, W.R.H. by NHMRC 1154457, and W.R.H. and L.E.H. by NHMRC 2012701.

Author information

Author notes

  1. These authors contributed equally: Mitch Ganley, Lauren E. Holz.

  2. These authors jointly supervised this work: Gavin F. Painter, Ian F. Hermans, William R. Heath.

Authors and Affiliations

  1. Ferrier Research Institute, Victoria University of Wellington, Lower Hutt, New Zealand

    Mitch Ganley, Sarah L. Draper, Susanna T. S. Chan, Regan J. Anderson, Benjamin J. Compton, Andrew J. Marshall & Gavin F. Painter

  2. Maurice Wilkins Centre for Molecular Biodiscovery, Auckland, New Zealand

    Mitch Ganley, David F. Ackerley, Gavin F. Painter & Ian F. Hermans

  3. Department of Microbiology and Immunology, The Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria, Australia

    Lauren E. Holz, Maria N. de Menezes, Yu Cheng Chua, Zhengyu Ge, Calvin Xu, Daniel Fernandez-Ruiz & William R. Heath

  4. Malaghan Institute of Medical Research, Wellington, New Zealand

    Jordan J. Minnell, Olivia K. Burn, Kathryn J. Farrand, John C. Mamum & Ian F. Hermans

  5. Infection and Immunity Program and Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia

    Kean Chan Yew Poa, Stephanie Gras, Jérôme Le Nours & Jamie Rossjohn

  6. Centenary Institute and University of Sydney, AW Morrow Gastroenterology and Liver Centre, Royal Prince Alfred Hospital, Sydney, New South Wales, Australia

    Kieran English & Patrick Bertolino

  7. School of BioSciences, University of Melbourne, Parkville, Victoria, Australia

    Anton Cozijnsen & Geoffrey I. McFadden

  8. Department of Immunology and Infectious Disease, John Curtin School of Medical Research, Australian National University, Canberra, Australian Capital Territory, Australia

    Ian A. Cockburn

  9. Shionogi Global Infectious Diseases Division, Institute of Tropical Medicine, Nagasaki University, Sakamoto, Nagasaki, Japan

    Katsuyuki Yui

  10. Department of Biochemistry and Chemistry, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Victoria, Australia

    Stephanie Gras

  11. Institute of Infection and Immunity, Cardiff University School of Medicine, Cardiff, UK

    Jamie Rossjohn

  12. School of Biological Sciences, Victoria University of Wellington, Wellington, New Zealand

    David F. Ackerley

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Contributions

W.R.H., I.F.H, G.F.P, D.F.-R., L.E.H., D.F.A. and M.G. conceived the idea and designed the outline of the research. A.C., I.A.C., K.Y. and G.I.M. provided sporozoites for infection studies. K.C.Y.P., J.L.N. and J.R. carried out affinity measurements of 2C12 TCR–CD1d–αGCB by surface plasmon resonance. W.R.H., I.F.H., G.F.P., L.E.H. and M.G. wrote the manuscript. J.L.N., S.L.D., S.T.S.C, R.J.A., B.J.C. A.J.M. O.K.B. and J.J.M. wrote Methods sections. All authors contributed to reviewing and editing the manuscript and to relevant discussions. M.G., K.J.F., O.K.B., J.C.M., M.M., L.E.H., J.J.M., Y.C.C. and Z.G. vaccinated mice, analyzed T cell responses and examined protection. S.L.D., M.G., J.J.M., S.T.S.C, R.J.A., B.J.C. and A.M. prepared vaccines and vaccine components. S.G. produced Kb tetramers. C.X. produced CD1d–PBS-44 tetramers. K.E. and P.B. contributed to analysis of liver DC.

Corresponding authors

Correspondence to Gavin F. Painter, Ian F. Hermans or William R. Heath.

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Competing interests

M.G., L.E.H., R.J.A., B.J.C., A.J.M. I.F.H., W.R.H. and G.F.P. are inventors on a patent application (WO2023121483A1) submitted by Victoria University of Wellington subsidiary Victoria Link Limited that covers the production of tissue-resident memory T cells with mRNA vaccines.

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Peer review information

Nature Immunology thanks Rafick Sekaly and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Primary Handling Editor: Jamie D. K. Wilson, in collaboration with the Nature Immunology team.

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Extended data

Extended Data Fig. 1 Vaccine-induced activation of splenic NKT cells.

Male B6 mice were transferred OT-I.CD45.1 cells one day before vaccination with mOVA vaccine alone, or mOVA with 80 pmol of indicated adjuvants. Data are combined from two independent experiments, giving n = 10 mice per group with the exception of the mOVA + αGCB group which contains 9 mice. a, Example flow cytometry plots showing expression of the TCR on splenic NKT cells at day 28. Gating shown in Supplementary data Fig. 4. b, Examples of CD69 (left) and PD-1 (right) expression on splenic NKT cells. c, Percentage TCRβ+ CD1d-tetramer+ spleen cells as gated in (a). Data are displayed as mean ± s.e.m. and individual mice (circles) and compared to the mOVA + αGC group by one-way ANOVA analysis with Tukey’s multiple comparison post-test. d, Geometric mean fluorescence intensity of PD-1 on NKT cells. Data are displayed as mean ± s.e.m. from two independent experiments and compared to the mOVA + αGC group by one-way ANOVA analysis with Tukey’s multiple comparison post-test. ****P < 0.0001.

Source data

Extended Data Fig. 2 Type I NKT 2C12 TCR binding affinity for mouse CD1d-αGCB.

Affinity measurement of soluble mouse type I NKT 2C12 TCR to mCD1d-αGCB using surface plasmon resonance (SPR). a, Equilibrium curves representative of one experiment performed in duplicate. b, Kd values are derived from duplicate runs from n = 3 independent experiments. Error bars on lower graph show mean ± SD.

Source data

Extended Data Fig. 3 Effect of vaccination on expression of costimulatory molecules on liver DC.

Female B6 mice were vaccinated with mOVA vaccine alone or mOVA with adjuvants αGC or αGCB. Livers were harvested 24 h later and DC (CD11c+ MHC II+ cells) assessed for relative expression of CD80 (a) and CD86 (b). Data are expressed as percent of maximum (% of max) determined by dividing the GMFI of each sample by the maximum mean GMFI of the highest mean of all groups from each experiment. Each circle depicts the values of an individual mouse (n = 10 biologically distinct samples combined from two independent experiments). Lines show mean ± s.e.m. Data (a, b) were compared by a one-way ANOVA with Tukey’s multiple comparison post-test (a, b). ****P < 0.0001.

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Extended Data Fig. 4 The role of cytokines in responses to mRNA vaccination.

a, Male B6 mice were transferred OT-I.CD45.1 cells one day before vaccination with mOVA + αGCB. Groups of these mice were injected with mAb specific for blocking either IL-12p35, IL-15, GM-CSF or IFN-γR1, or with an isotype control mAb on days 0, 1 and 3 and then left to day 30 before OT-I memory T cell subsets were quantified in the liver. Gating parameters for OT-I cells are described in Supplementary data Fig. 1. Data are expressed as mean cell count ± s.e.m. for each subset. Individual values for each mouse are shown in Supplementary Fig. 8a. b, Female B6 mice or IL-15−/− mice were transferred OT-I.CD45.1 cells one day before vaccination with mOVA + αGCB., and 36 days later OT-I memory T cell subsets were quantified in the liver. Mean cell count ± s.e.m. for each subset shown. Individual values for each mouse are shown in Supplementary Fig. 8b. c, Male B6 mice were OT-I.CD45.1 cells one day before vaccination mOVA + αGCB. Groups of these mice were injected with specific mAb for blocking IFNαR1 or an isotype control on days 0, 1 and 3 and then left to day 30 before OT-I memory T cell subsets were quantified in the liver. Mean cell count ± s.e.m. for each subset shown. Individual values for each mouse are shown in Supplementary data 8c. Data in a were log-transformed and compared to the isotype group by one-way ANOVA with Tukey’s multiple comparison post-test. Data in b and c were log-transformed and compared by two-sided unpaired Student’s t tests. Two independent experiments were performed for each panel (a,b, n = 10 mice per group; c, n = 7 mice per group). ****P < 0.0001. Bars indicating significance are coloured to correspond to each memory T cell subset.

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Extended Data Fig. 5 Assessment of total anti-OVA IgG in sera 3 weeks following a prime-boost schedule.

Male B6 mice were vaccinated with mOVA vaccine alone (containing 5 µg mRNA), or mOVA adjuvanted with αGC or αGCB, on days 0 and 21, with sera collected 3 weeks after the boost analysed for total anti-OVA IgG by ELISA. Two independent experiments were combined to give 10 mice per group with the exception of the αGC group which contains 11 mice. Geometric mean ± 95 % confidence intervals for EC50 values are shown. Data were log-transformed and compared by one-way ANOVA with Tukey’s multiple comparison post-test. ****P < 0.0001.

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Ganley, M., Holz, L.E., Minnell, J.J. et al. mRNA vaccine against malaria tailored for liver-resident memory T cells. Nat Immunol 24, 1487–1498 (2023). https://doi.org/10.1038/s41590-023-01562-6

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