Direct interaction of whole-inactivated influenza A and pneumococcal vaccines enhances influenza-specific immunity

Abstract

The upper respiratory tract is continuously exposed to a vast array of potentially pathogenic viruses and bacteria. Influenza A virus (IAV) has particular synergism with the commensal bacterium Streptococcus pneumoniae in this niche, and co-infection exacerbates pathogenicity and causes significant mortality. However, it is not known whether this synergism is associated with a direct interaction between the two pathogens. We have previously reported that co-administration of a whole-inactivated IAV vaccine (γ-Flu) with a whole-inactivated pneumococcal vaccine (γ-PN) enhances pneumococcal-specific responses. In this study, we show that mucosal co-administration of γ-Flu and γ-PN similarly augments IAV-specific immunity, particularly tissue-resident memory cell responses in the lung. In addition, our in vitro analysis revealed that S. pneumoniae directly interacts with both γ-Flu and with live IAV, facilitating increased uptake by macrophages as well as increased infection of epithelial cells by IAV. These observations provide an additional explanation for the synergistic pathogenicity of IAV and S. pneumoniae, as well as heralding the prospect of exploiting the phenomenon to develop better vaccine strategies for both pathogens.

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Fig. 1: Enhanced protection against drifted and heterosubtypic IAV challenge following co-vaccination with γ-Flu + γ-PN(ΔPsaA).
Fig. 2: In vitro neutralization of A/PR8 by vaccine-induced antibodies.
Fig. 3: IAV-specific T-cell populations in peripheral blood and secondary lymphoid organs.
Fig. 4: The magnitude of the IAV-specific CD8+ T-cell response in the lung is enhanced by co-vaccination.
Fig. 5: The presence of γ-PN(ΔPsaA) enhances the uptake of IAV by the THP-1 and MDCK cell lines.
Fig. 6: Direct association of γ-Flu and γ-PN(ΔPsaA) whole-inactivated vaccines.

Data availability

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

References

  1. 1.

    Kuss, S. K. et al. Intestinal microbiota promote enteric virus replication and systemic pathogenesis. Science 334, 249–252 (2011).

  2. 2.

    Kane, M. et al. Successful transmission of a retrovirus depends on the commensal microbiota. Science 334, 245–249 (2011).

  3. 3.

    Madhi, S. A., Klugman, K. P. & The Vaccine Trialist Group A role for Streptococcus pneumoniae in virus-associated pneumonia. Nat. Med. 10, 811–813 (2004).

  4. 4.

    Johnson, P. T. & Hoverman, J. T. Parasite diversity and coinfection determine pathogen infection success and host fitness. Proc. Natl Acad. Sci. USA 109, 9006–9011 (2012).

  5. 5.

    McCullers, J. A. Insights into the interaction between influenza virus and pneumococcus. Clin. Microbiol. Rev. 19, 571–582 (2006).

  6. 6.

    Blyth, C. C. et al. The impact of bacterial and viral co-infection in severe influenza. Influenza Other Respir. Viruses 7, 168–176 (2013).

  7. 7.

    Nelson, G. E., Gershman, K. A., Swerdlow, D. L., Beall, B. W. & Moore, M. R. Invasive pneumococcal disease and pandemic (H1N1) 2009, Denver, Colorado, USA. Emerg. Infect. Dis. 18, 208–216 (2012).

  8. 8.

    Chien, Y. W., Klugman, K. P. & Morens, D. M. Bacterial pathogens and death during the 1918 influenza pandemic. N. Engl. J. Med. 361, 2582–2583 (2009).

  9. 9.

    Zhang, Y. Y. et al. Comparison of dual influenza and pneumococcal polysaccharide vaccination with influenza vaccination alone for preventing pneumonia and reducing mortality among the elderly: a meta-analysis. Hum. Vaccin. Immunother. 12, 3056–3064 (2016).

  10. 10.

    Chan, T. C. et al. Prevention of mortality and pneumonia among nursing home older adults by dual pneumococcal and seasonal influenza vaccination during a pandemic caused by novel pandemic influenza A (H1N1). J. Am. Med. Dir. Assoc. 13, 698–703 (2012).

  11. 11.

    Krammer, F. & Palese, P. Advances in the development of influenza virus vaccines. Nat. Rev. Drug Discov. 14, 167–182 (2015).

  12. 12.

    Alsharifi, M. et al. Intranasal flu vaccine protective against seasonal and H5N1 avian influenza infections. PLoS ONE 4, e5336 (2009).

  13. 13.

    Furuya, Y. et al. Cytotoxic T cells are the predominant players providing cross-protective immunity induced by γ-irradiated influenza A viruses. J. Virol. 84, 4212–4221 (2010).

  14. 14.

    David, S. C. et al. The effect of gamma-irradiation conditions on the immunogenicity of whole-inactivated Influenza A virus vaccine. Vaccine 35, 1071–1079 (2017).

  15. 15.

    Babb, R. et al. Enhanced protective responses to a serotype-independent pneumococcal vaccine when combined with an inactivated influenza vaccine. Clin. Sci. 131, 169–180 (2017).

  16. 16.

    Gerlach, C. et al. The chemokine receptor CX3CR1 defines three antigen-experienced CD8 T cell subsets with distinct roles in immune surveillance and homeostasis. Immunity 45, 1270–1284 (2016).

  17. 17.

    Mueller, S. N., Gebhardt, T., Carbone, F. R. & Heath, W. R. Memory T cell subsets, migration patterns, and tissue residence. Annu Rev. Immunol. 31, 137–161 (2013).

  18. 18.

    Schenkel, J. M. & Masopust, D. Tissue-resident memory T cells. Immunity 41, 886–897 (2014).

  19. 19.

    Mackay, L. K. et al. The developmental pathway for CD103+CD8+ tissue-resident memory T cells of skin. Nat. Immunol. 14, 1294–1301 (2013).

  20. 20.

    Cibrian, D. & Sanchez-Madrid, F. CD69: from activation marker to metabolic gatekeeper. Eur. J. Immunol. 47, 946–953 (2017).

  21. 21.

    Gunn, M. D., Nelken, N. A., Liao, X. & Williams, L. T. Monocyte chemoattractant protein-1 is sufficient for the chemotaxis of monocytes and lymphocytes in transgenic mice but requires an additional stimulus for inflammatory activation. J. Immunol. 158, 376–383 (1997).

  22. 22.

    Lindell, D. M., Lane, T. E. & Lukacs, N. W. CXCL10/CXCR3-mediated responses promote immunity to respiratory syncytial virus infection by augmenting dendritic cell and CD8+ T cell efficacy. Eur. J. Immunol. 38, 2168–2179 (2008).

  23. 23.

    Fadel, S. A., Bromley, S. K., Medoff, B. D. & Luster, A. D. CXCR3-deficiency protects influenza-infected CCR5-deficient mice from mortality. Eur. J. Immunol. 38, 3376–3387 (2008).

  24. 24.

    Wu, T. et al. Lung-resident memory CD8 T cells (TRM) are indispensable for optimal cross-protection against pulmonary virus infection. J. Leukoc. Biol. 95, 215–224 (2014).

  25. 25.

    Zens, K. D., Chen, J. K. & Farber, D. L. Vaccine-generated lung tissue-resident memory T cells provide heterosubtypic protection to influenza infection. JCI Insight 1, e85832 (2016).

  26. 26.

    Skon, C. N. et al. Transcriptional downregulation of S1pr1 is required for the establishment of resident memory CD8+ T cells. Nat. Immunol. 14, 1285–1293 (2013).

  27. 27.

    Casey, K. A. et al. Antigen-independent differentiation and maintenance of effector-like resident memory T cells in tissues. J. Immunol. 188, 4866–4875 (2012).

  28. 28.

    Mueller, S. N. & Mackay, L. K. Tissue-resident memory T cells: local specialists in immune defence. Nat. Rev. Immunol. 16, 79–89 (2016).

  29. 29.

    Khan, T. N., Mooster, J. L., Kilgore, A. M., Osborn, J. F. & Nolz, J. C. Local antigen in nonlymphoid tissue promotes resident memory CD8+ T cell formation during viral infection. J. Exp. Med. 213, 951–966 (2016).

  30. 30.

    Williams, A. E., Jose, R. J., Brown, J. S. & Chambers, R. C. Enhanced inflammation in aged mice following infection with Streptococcus pneumoniae is associated with decreased IL-10 and augmented chemokine production. Am. J. Physiol. Lung Cell Mol. Physiol. 308, L539–L549 (2015).

  31. 31.

    Neill, D. R. et al. Density and duration of pneumococcal carriage is maintained by transforming growth factor beta1 and T regulatory cells. Am. J. Respir. Crit. Care Med. 189, 1250–1259 (2014).

  32. 32.

    Slutter, B. et al. Dynamics of influenza-induced lung-resident memory T cells underlie waning heterosubtypic immunity. Sci. Immunol. 2, eaag2031 (2017).

  33. 33.

    Laidlaw, B. J. et al. CD4+ T cell help guides formation of CD103+ lung-resident memory CD8+ T cells during influenza viral infection. Immunity 41, 633–645 (2014).

  34. 34.

    Nakanishi, Y., Lu, B., Gerard, C. & Iwasaki, A. CD8+ T lymphocyte mobilization to virus-infected tissue requires CD4+ T-cell help. Nature 462, 510–513 (2009).

  35. 35.

    Appay, V. et al. Characterization of CD4+ CTLs ex vivo. J. Immunol. 168, 5954–5958 (2002).

  36. 36.

    Brown, D. M., Lee, S., Garcia-Hernandez Mde, L. & Swain, S. L. Multifunctional CD4 cells expressing gamma interferon and perforin mediate protection against lethal influenza virus infection. J. Virol. 86, 6792–6803 (2012).

  37. 37.

    Wilkinson, T. M. et al. Preexisting influenza-specific CD4+ T cells correlate with disease protection against influenza challenge in humans. Nat. Med. 18, 274–280 (2012).

  38. 38.

    Pozzi, L. A., Maciaszek, J. W. & Rock, K. L. Both dendritic cells and macrophages can stimulate naive CD8 T cells in vivo to proliferate, develop effector function, and differentiate into memory cells. J. Immunol. 175, 2071–2081 (2005).

  39. 39.

    Chatziandreou, N. et al. Macrophage death following influenza vaccination initiates the inflammatory response that promotes dendritic cell function in the draining lymph node. Cell Rep. 18, 2427–2440 (2017).

  40. 40.

    Hoffmann, J. et al. Viral and bacterial co-infection in severe pneumonia triggers innate immune responses and specifically enhances IP-10: a translational study. Sci. Rep. 6, 38532 (2016).

  41. 41.

    Stark, J. M., Stark, M. A., Colasurdo, G. N. & LeVine, A. M. Decreased bacterial clearance from the lungs of mice following primary respiratory syncytial virus infection. J. Med. Virol. 78, 829–838 (2006).

  42. 42.

    Alymova, I. V. et al. The novel parainfluenza virus hemagglutinin-neuraminidase inhibitor BCX 2798 prevents lethal synergism between a paramyxovirus and Streptococcus pneumoniae. Antimicrob. Agents Chemother. 49, 398–405 (2005).

  43. 43.

    Kukavica-Ibrulj, I. et al. Infection with human metapneumovirus predisposes mice to severe neumococcal pneumonia. J. Virol. 83, 1341–1349 (2009).

  44. 44.

    Okamoto, S. et al. The Streptococcus pyogenes capsule is required for adhesion of bacteria to virus-infected alveolar epithelial cells and lethal bacterial-viral superinfection. Infect. Immun. 72, 6068–6075 (2004).

  45. 45.

    Wang, Y. et al. Capsular sialic acid of Streptococcus suis serotype 2 binds to swine influenza virus and enhances bacterial interactions with virus-infected tracheal epithelial cells. Infect. Immun. 81, 4498–4508 (2013).

  46. 46.

    Hosaka, Y. et al. Binding of influenza type A viruses to group B Streptococcus and haemagglutination by virus-bound bacteria. J. Electron Microsc. 49, 765–773 (2000).

  47. 47.

    Sheffield, F. W., Smith, W. & Belyavin, G. Purification of influenza virus by red-cell adsorption and elution. Br. J. Exp. Pathol. 35, 214–222 (1954).

  48. 48.

    Cottey, R., Rowe, C. A. & Bender, B. S. Influenza Virus. Curr. Protoc. Immunol. 42, 19.11.1–19.11.32 (2001).

  49. 49.

    Babb, R. et al. Intranasal vaccination with gamma-irradiated Streptococcus pneumoniae whole-cell vaccine provides serotype-independent protection mediated by B-cells and innate IL-17 responses. Clin. Sci. 130, 697–710 (2016).

  50. 50.

    Harvey, R. M., Ogunniyi, A. D., Chen, A. Y. & Paton, J. C. Pneumolysin with low hemolytic activity confers an early growth advantage to Streptococcus pneumoniae in the blood. Infect. Immunol. 79, 4122–4130 (2011).

  51. 51.

    WHO Expert Committee on Biological Standardization (World Health Organization, 2005).

  52. 52.

    Hogquist, K. A. et al. T cell receptor antagonist peptides induce positive selection. Cell 76, 17–27 (1994).

  53. 53.

    Chan, J., Babb, R., David, S. C., McColl, S. R. & Alsharifi, M. Vaccine-induced antibody responses prevent the induction of interferon type I responses upon a homotypic live virus challenge. Scand. J. Immunol. 83, 165–173 (2016).

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Acknowledgements

The authors thank Adelaide University Microscopy (Adelaide, Australia) for assistance in sample preparation and operation of the FEI Tecnai G2 Spirit TEM. We also acknowledge the following funding sources supporting this research: an Australian Institute of Nuclear Science and Engineering (AINSE) Research Award (ALNGRA15517; to M.A.); an Australian Postgraduate Award and Gamma Vaccines Pty Ltd research sponsorship (to S.C.D.); and a National Health and Medical Research Council Senior Principal Research Fellowship (awarded to J.C.P.).

Author information

M.A., S.C.D. and J.C.P. conceived and designed the study. S.C.D., T.N., T.T., J.J.W., E.V.S., Z.L. and J.D. performed the experiments. S.C.D. and T.N. performed statistical analysis. S.C.D. and M.A. wrote the manuscript. T.R.H., I.C. and S.R.M. assisted in experimental design, preparation of the manuscript and provided reagents. M.A. and J.C.P. supervised the study.

Correspondence to Mohammed Alsharifi.

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Competing interests

M.A. is head of the vaccine research group at the University of Adelaide and the Chief Scientific Officer of Gamma Vaccines Pty Ltd and Director of GPN Vaccines Pty Ltd, J.C.P. is a Director of GPN Vaccines Pty Ltd, and T.R.H. is the Executive Chairman of Gamma Vaccines Pty Ltd and GPN Vaccines Pty Ltd. This does not alter adherence to policies on sharing data and materials. Both Gamma Vaccines Pty Ltd and GPN Vaccines Pty Ltd have no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Supplementary Figures 1–3, Tables 1 and 2, and Flow Cytometry Gating Strategies 1–5.

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David, S.C., Norton, T., Tyllis, T. et al. Direct interaction of whole-inactivated influenza A and pneumococcal vaccines enhances influenza-specific immunity. Nat Microbiol 4, 1316–1327 (2019). https://doi.org/10.1038/s41564-019-0443-4

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