Single-cell analysis reveals lasting immunological consequences of influenza infection and respiratory immunization in the pig lung

PLoS Pathog. 2024 Jul 18;20(7):e1011910. doi: 10.1371/journal.ppat.1011910. eCollection 2024 Jul.

Abstract

The pig is a natural host for influenza viruses and integrally involved in virus evolution through interspecies transmissions between humans and swine. Swine have many physiological, anatomical, and immunological similarities to humans, and are an excellent model for human influenza. Here, we employed single cell RNA-sequencing (scRNA-seq) and flow cytometry to characterize the major leukocyte subsets in bronchoalveolar lavage (BAL), twenty-one days after H1N1pdm09 infection or respiratory immunization with an adenoviral vector vaccine expressing hemagglutinin and nucleoprotein with or without IL-1β. Mapping scRNA-seq clusters from BAL onto those previously described in peripheral blood facilitated annotation and highlighted differences between tissue resident and circulating immune cells. ScRNA-seq data and functional assays revealed lasting impacts of immune challenge on BAL populations. First, mucosal administration of IL-1β reduced the number of functionally active Treg cells. Second, influenza infection upregulated IFI6 in BAL cells and decreased their susceptibility to virus replication in vitro. Our data provide a reference map of porcine BAL cells and reveal lasting immunological consequences of influenza infection and respiratory immunization in a highly relevant large animal model for respiratory virus infection.

MeSH terms

  • Animals
  • Bronchoalveolar Lavage Fluid / immunology
  • Bronchoalveolar Lavage Fluid / virology
  • Immunization
  • Influenza A Virus, H1N1 Subtype* / immunology
  • Influenza Vaccines* / immunology
  • Lung* / immunology
  • Lung* / virology
  • Orthomyxoviridae Infections* / immunology
  • Orthomyxoviridae Infections* / virology
  • Single-Cell Analysis*
  • Swine

Substances

  • Influenza Vaccines

Grants and funding

This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC) Strategic Program and Core Capability Grants to The Pirbright Institute (BBS/E/I/00007031, BBS/E/I/00007037, BBS/E/PI/230001A, The Pirbright Institute Development Grant [IDG], 2021) and BBSRC grants to the Babraham Institute (BBS/E/B/000C0427, Babraham Institute Development [IDG] Grant 2021, Babraham Institute Core Capability Grant). A VetBioNet award, under the European Union’s Horizon 2020 research and innovation programme, grant agreement No 731014 (Ref: VBN_20_49) was granted to MT. The authors would also like to acknowledge the Bioinformatics, Sequencing, Proteomics and Pirbright Flow Cytometry facility and support through the Core capability grant (BBS/E/I/00007039, and BBS/E/PI/23NB0003). AR is supported by an MRC Career Development Award (MR/W016303/1). The Biological Research Facility and Immunological Toolbox (generation of CD161 mAb) are supported by funding from UKRI-BBSRC (BB/CCG2270/1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.