Programming the magnitude and persistence of antibody responses with innate immunity

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Many successful vaccines induce persistent antibody responses that can last a lifetime. The mechanisms by which they do so remain unclear, but emerging evidence indicates that they activate dendritic cells via Toll-like receptors (TLRs)1,2. For example, the yellow fever vaccine YF-17D, one of the most successful empiric vaccines ever developed3, activates dendritic cells via multiple TLRs to stimulate proinflammatory cytokines4,5. Triggering specific combinations of TLRs in dendritic cells can induce synergistic production of cytokines6, which results in enhanced T-cell responses, but its impact on antibody responses remain unknown. Learning the critical parameters of innate immunity that program such antibody responses remains a major challenge in vaccinology. Here we demonstrate that immunization of mice with synthetic nanoparticles containing antigens plus ligands that signal through TLR4 and TLR7 induces synergistic increases in antigen-specific, neutralizing antibodies compared to immunization with nanoparticles containing antigens plus a single TLR ligand. Consistent with this there was enhanced persistence of germinal centres and of plasma-cell responses, which persisted in the lymph nodes for >1.5 years. Surprisingly, there was no enhancement of the early short-lived plasma-cell response relative to that observed with single TLR ligands. Molecular profiling of activated B cells, isolated 7 days after immunization, indicated that there was early programming towards B-cell memory. Antibody responses were dependent on direct triggering of both TLRs on B cells and dendritic cells, as well as on T-cell help. Immunization protected completely against lethal avian and swine influenza virus strains in mice, and induced robust immunity against pandemic H1N1 influenza in rhesus macaques.

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Figure 1: Combination of MPL and R837 in PLGA nanoparticles mediates synergistic enhancement of antibody responses against H5N1-influenza-derived HA.
Figure 2: Synergistic enhancement of antibody responses is dependent on the presence of TLRs on B cells.
Figure 3: Immunization with nanoparticles containing MPL and R837 induces persistent germinal centres and long-lived antibody-forming cells in draining lymph nodes.
Figure 4: Immunization of rhesus macaques with 2009 pandemic H1N1 influenza A, whole inactivated virus and nanoparticles containing MPL+R837 or MPL+R848 induces robust humoral immune responses.

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Gene Expression Omnibus

Data deposits

All microarray data are deposited in the Gene Expression Omnibus under accession number GSE25677.


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We thank R. Ahmed and B. Rouse for discussion and comments on the manuscript. We thank B. Norris for assistance with FACS analysis, and H. Oluoch for assistance with cryostat sectioning. The work in the laboratory of B.P. was supported by grants U54AI057157, R37AI48638, R01DK057665, U19AI057266, HHSN266200700006C, NO1 AI50025 and U19AI090023 from the National Institutes of Health and a grant from the Bill & Melinda Gates Foundation. Work in A.G.-S. laboratories was partly funded by grants HHSN266200700010C, U54AI57158) and U01AI070469 from the National Institutes of Health.

Author information

S.P.K. and B.P. designed the study, planned the experiments and analysed the data. B.P. and S.P.K. wrote the manuscript. S.P.K., I.S. and B.P. designed and performed the H1N1 vaccine studies in mice and primates. D.K. assisted with the H1N1 vaccine studies in mice and primates. R.A.A., A.G.-S. and J.S. designed and performed the neutralization assays and challenge experiments with H5N1 vaccine studies in mice. T.H. and R.R. assisted with experiments. H.I.N. performed the microarray analysis. S.S. and M.A. designed and carried out the SPR-based avidity experiments. M.K. assisted with design and execution of mice and non-human primate experiments. N.M. assisted with design of formulations. J.J. assisted with immunohistochemistry and design of experiments. R.J.H. expressed and purified the recombinant H5HA protein. R.C. helped plan and design the H1N1 vaccine study in mice and primates.

Correspondence to Bali Pulendran.

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Kasturi, S., Skountzou, I., Albrecht, R. et al. Programming the magnitude and persistence of antibody responses with innate immunity. Nature 470, 543–547 (2011) doi:10.1038/nature09737

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