Antigens reversibly conjugated to a polymeric glyco-adjuvant induce protective humoral and cellular immunity

Abstract

Fully effective vaccines for complex infections must elicit a diverse repertoire of antibodies (humoral immunity) and CD8+ T-cell responses (cellular immunity). Here, we present a synthetic glyco-adjuvant named p(Man–TLR7), which, when conjugated to antigens, elicits robust humoral and cellular immunity. p(Man–TLR7) is a random copolymer composed of monomers that either target dendritic cells (DCs) via mannose-binding receptors or activate DCs via Toll-like receptor 7 (TLR7). Protein antigens are conjugated to p(Man–TLR7) via a self-immolative linkage that releases chemically unmodified antigen after endocytosis, thus amplifying antigen presentation to T cells. Studies with ovalbumin (OVA)–p(Man–TLR7) conjugates demonstrate that OVA–p(Man–TLR7) generates greater humoral and cellular immunity than OVA conjugated to polymers lacking either mannose targeting or TLR7 ligand. We show significant enhancement of Plasmodium falciparum-derived circumsporozoite protein (CSP)-specific T-cell responses, expansion in the breadth of the αCSP IgG response and increased inhibition of sporozoite invasion into hepatocytes with CSP–p(Man–TLR7) when compared with CSP formulated with MPLA/QS-21-loaded liposomes—the adjuvant used in the most clinically advanced malaria vaccine. We conclude that our antigen–p(Man–TLR7) platform offers a strategy to enhance the immunogenicity of protein subunit vaccines.

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Fig. 1: p(Man–TLR7) is a potent activator of DCs and OVA–p(Man–TLR7) is internalized via mannose-binding receptors.
Fig. 2: p(Man–TLR7) targets lymph nodes and increases antigen delivery to multiple DC subsets via mannose-binding receptors.
Fig. 3: OVA–p(Man–TLR7) vaccination avoids systemic inflammation and enhances the magnitude and quality of OVA-specific humoral and T-cell responses.
Fig. 4: OVA–p(Man–TLR7) induces superior T-cell and humoral responses compared with other formulations composed of polymeric TLR7.
Fig. 5: CSP–p(Man–TLR7) induces superior CSP-specific CD4+ and CD8+ T-cell response compared with CSP formulated with leading adjuvants.
Fig. 6: CSP–p(Man–TLR7) increases CSP-specific IgGs that are specific for a wide range of CSP epitopes and reduces malaria parasite burden in human hepatocytes.

Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank the Flow Cytometry Core Facility of EPFL for technical assistance, and Dr E. Simeoni of EPFL for discussion of the research and guidance on animal work. D.S.W. was supported by a fellowship from the Whitaker Foundation. We would like to thank PATH for donating the CSP used in our studies. This study was supported by the School of Life Sciences, EPFL, and the University of Chicago. This work benefited from equipment and services from the CELIS cell culture core facility (Institut du Cerveau et de la Moelle Epinière, Paris), a platform supported through the ANR grants ANR-10-IAIHU-06 and ANR-11-INBS-0011-NeurATRIS. We are particularly grateful to D. Akbar for his assistance regarding automated fluorescence.

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J.A.H. and M.A.S. oversaw all research; D.S.W. conceptualized materials; D.S.W., M.M.R. and R.W. performed the synthesis; D.S.W., S.H. and J.A.H designed animal studies; D.S.W., S.H., M.A.S.B., L.J., G.D. and X.Q.T. performed animal studies; S.H., L.J., D.S.W. and M.K. designed and carried out in vitro studies; D.M., J.-F.F. and L.B.R. designed hepatic invasion studies; J.-F.F. and L.B.R. carried out hepatic invasion studies; D.S.W., S.H. and J.A.H. wrote the manuscript; all authors proofread the manuscript.

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Correspondence to Jeffrey A. Hubbell.

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The University of Chicago has filed for patent protection on the p(Man-TLRx) delivery platform, and J.A.H., S.H. and D.S.W. are named as inventors on these patents.

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Wilson, D.S., Hirosue, S., Raczy, M.M. et al. Antigens reversibly conjugated to a polymeric glyco-adjuvant induce protective humoral and cellular immunity. Nature Mater 18, 175–185 (2019). https://doi.org/10.1038/s41563-018-0256-5

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