Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Turbinate-homing IgA-secreting cells originate in the nasal lymphoid tissues

Nature volume 632, pages 637–646 (2024)Cite this article

Subjects

Abstract

Nasal vaccination elicits a humoral immune response that provides protection from airborne pathogens1, yet the origins and specific immune niches of antigen-specific IgA-secreting cells in the upper airways are unclear2. Here we define nasal glandular acinar structures and the turbinates as immunological niches that recruit IgA-secreting plasma cells from the nasal-associated lymphoid tissues (NALTs)3. Using intact organ imaging, we demonstrate that nasal vaccination induces B cell expansion in the subepithelial dome of the NALT, followed by invasion into commensal-bacteria-driven chronic germinal centres in a T cell-dependent manner. Initiation of the germinal centre response in the NALT requires pre-expansion of antigen-specific T cells, which interact with cognate B cells in interfollicular regions. NALT ablation and blockade of PSGL-1, which mediates interactions with endothelial cell selectins, demonstrated that NALT-derived IgA-expressing B cells home to the turbinate region through the circulation, where they are positioned primarily around glandular acinar structures. CCL28 expression was increased in the turbinates in response to vaccination and promoted homing of IgA+ B cells to this site. Thus, in response to nasal vaccination, the glandular acini and turbinates provide immunological niches that host NALT-derived IgA-secreting cells. These cellular events could be manipulated in vaccine design or in the treatment of upper airway allergic responses.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Nasal vaccination induces antigen-specific B cell expansion in the NALT SED.
Fig. 2: Pre-expansion of CD4 T cells is required for GC formation in the NALT.
Fig. 3: Antigen-specific B cells are recruited to the NTs in response to nasal vaccination.
Fig. 4: Antigen-specific B cells are recruited to the NTs in response to nasal vaccination.
Fig. 5: CCL28 promotes B cell homing to the turbinates.

Similar content being viewed by others

Data availability

The 10x scRNA-seq data and the bulk RNA-seq dataset have been deposited at the NCBI Gene Expression Omnibus database under SuperSeries GSE264643, under accession numbers GSE252018 and GSE264641, separately. All data supporting the findings of this study are available from the corresponding author on request. There are no restrictions on data availability. Source data are provided with this paper.

References

  1. Lavelle, E. C. & Ward, R. W. Mucosal vaccines—fortifying the frontiers. Nat. Rev. Immunol. 22, 236–250 (2021).

    Article  PubMed  PubMed Central  Google Scholar 

  2. Wellford, S. A. & Moseman, E. A. Olfactory immunology: the missing piece in airway and CNS defence. Nat. Rev. Immunol. https://doi.org/10.1038/s41577-023-00972-9 (2023).

  3. Bienenstock, J. & McDermott, M. R. Bronchus- and nasal-associated lymphoid tissues. Immunol. Rev. 206, 22–31 (2005).

    Article  PubMed  Google Scholar 

  4. Neutra, M. R., Frey, A. & Kraehenbuhl, J. P. Epithelial M cells: gateways for mucosal infection and immunization. Cell 86, 345–348 (1996).

    Article  CAS  PubMed  Google Scholar 

  5. Komban, R. J. et al. Activated Peyer’s patch B cells sample antigen directly from M cells in the subepithelial dome. Nat. Commun. 10, 2423 (2019).

    Article  ADS  PubMed  PubMed Central  Google Scholar 

  6. Kiyono, H. & Fukuyama, S. NALT- versus Peyer’s-patch-mediated mucosal immunity. Nat. Rev. Immunol. 4, 699–710 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Lund, F. E. & Randall, T. D. Scent of a vaccine. Science 373, 397–399 (2021).

    Article  ADS  CAS  PubMed  Google Scholar 

  8. Randall, T. D. in Mucosal Immunology 4th edn (eds Mestecky, J. et al.) 43–61, Ch. 4 (Academic, 2015).

  9. Victora, G. D. & Nussenzweig, M. C. Germinal centers. Annu. Rev. Immunol. 40, 413–442 (2022).

    Article  CAS  PubMed  Google Scholar 

  10. Biram, A. et al. BCR affinity differentially regulates colonization of the subepithelial dome and infiltration into germinal centers within Peyer’s patches. Nat. Immunol. 20, 482–492 (2019).

    Article  CAS  PubMed  Google Scholar 

  11. Reboldi, A. et al. IgA production requires B cell interaction with subepithelial dendritic cells in Peyer’s patches. Science 352, aaf4822 (2016).

    Article  PubMed  PubMed Central  Google Scholar 

  12. Bergqvist, P. et al. Re-utilization of germinal centers in multiple Peyer’s patches results in highly synchronized, oligoclonal, and affinity-matured gut IgA responses. Mucosal Immunol. 6, 122 (2012).

    Article  PubMed  Google Scholar 

  13. Hartwell, B. L. et al. Intranasal vaccination with lipid-conjugated immunogens promotes antigen transmucosal uptake to drive mucosal and systemic immunity. Sci. Transl. Med. 14, eabn1413 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Topol, E. J. & Iwasaki, A. Operation nasal vaccine—lightning speed to counter COVID-19. Sci. Immunol. 7, eadd9947 (2022).

    Article  PubMed  Google Scholar 

  15. Mao, T. et al. Unadjuvanted intranasal spike vaccine elicits protective mucosal immunity against sarbecoviruses. Science 378, eabo2523 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Wu, C.-T. et al. SARS-CoV-2 replication in airway epithelia requires motile cilia and microvillar reprogramming. Cell 186, 112–130 (2023).

    Article  CAS  PubMed  Google Scholar 

  17. van Riel, D., Verdijk, R. & Kuiken, T. The olfactory nerve: a shortcut for influenza and other viral diseases into the central nervous system. J. Pathol. 235, 277–287 (2015).

    Article  PubMed  Google Scholar 

  18. Chamanza, R. & Wright, J. A. A review of the comparative anatomy, histology, physiology and pathology of the nasal cavity of rats, mice, dogs and non-human primates. Relevance to inhalation toxicology and human health risk assessment. J. Comp. Pathol. 153, 287–314 (2015).

    Article  CAS  PubMed  Google Scholar 

  19. Wellford, S. A. et al. Mucosal plasma cells are required to protect the upper airway and brain from infection. Immunity https://doi.org/10.1016/j.immuni.2022.08.017 (2022).

  20. Harkema, J. R., Carey, S. A. & Wagner, J. G. The nose revisited: a brief review of the comparative structure, function, and toxicologic pathology of the nasal epithelium. Toxicol. Pathol. 34, 252–269 (2006).

    Article  CAS  PubMed  Google Scholar 

  21. Ualiyeva, S. et al. A nasal cell atlas reveals heterogeneity of tuft cells and their role in directing olfactory stem cell proliferation. Sci. Immunol. 9, eabq4341 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Bates, J. T. Naïve CD4 T cell activation in the nasal-associated lymphoid tissue following intranasal immunization with a flagellin-based subunit vaccine. Int. J. Mol. Sci. 23, 15572 (2022).

  23. Bedford, J. G. et al. Unresponsiveness to inhaled antigen is governed by conventional dendritic cells and overridden during infection by monocytes. Sci. Immunol. 5, eabb5439 (2020).

    Article  CAS  PubMed  Google Scholar 

  24. Yang Shih, T.-A., Meffre, E., Roederer, M. & Nussenzweig, M. C. Role of BCR affinity in T cell-dependent antibody responses in vivo. Nat. Immunol. 3, 570–575 (2002).

    Article  CAS  Google Scholar 

  25. Biram, A. et al. Bacterial infection disrupts established germinal center reactions through monocyte recruitment and impaired metabolic adaptation. Immunity 55, 442–458 (2022).

    Article  CAS  PubMed  Google Scholar 

  26. Stoler-Barak, L. et al. B cell dissemination patterns during the germinal center reaction revealed by whole-organ imaging. J. Exp. Med. 216, 2515–2530 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Bemark, M., Pitcher, M. J., Dionisi, C. & Spencer, J. Gut-associated lymphoid tissue: a microbiota-driven hub of B cell immunity. Trends Immunol. 45, 211–223 (2024).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Okada, T. et al. Antigen-engaged B cells undergo chemotaxis toward the T zone and form motile conjugates with helper T cells. PLoS Biol. 3, e150 (2005).

    Article  PubMed  PubMed Central  Google Scholar 

  29. Schwickert, T. A. et al. A dynamic T cell-limited checkpoint regulates affinity-dependent B cell entry into the germinal center. J. Exp. Med. 208, 1243–1252 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Roco, J. A. et al. Class-switch recombination occurs infrequently in germinal centers. Immunity 51, 337–350 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Grenov, A., Hezroni, H., Lasman, L., Hanna, J. H. & Shulman, Z. YTHDF2 suppresses the plasmablast genetic program and promotes germinal center formation. Cell Rep. 39, 110778 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Biram, A. et al. B cell diversification is uncoupled from SAP-mediated selection forces in chronic germinal centers within Peyer’s patches. Cell Rep. 30, 1910–1922 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Tsuji, M. et al. Requirement for lymphoid tissue-inducer cells in isolated follicle formation and T cell-independent immunoglobulin A generation in the gut. Immunity 29, 261–271 (2008).

    Article  CAS  PubMed  Google Scholar 

  34. Bergqvist, P., Stensson, A., Lycke, N. Y. & Bemark, M. T cell-independent IgA class switch recombination is restricted to the GALT and occurs prior to manifest germinal center formation. J. Immunol. 184, 3545–3553 (2010).

    Article  CAS  PubMed  Google Scholar 

  35. Bergqvist, P., Gärdby, E., Stensson, A., Bemark, M. & Lycke, N. Y. Gut IgA class switch recombination in the absence of CD40 does not occur in the lamina propria and is independent of germinal centers. J. Immunol. 177, 7772–7783 (2006).

    Article  CAS  PubMed  Google Scholar 

  36. Mora, J. R. & von Andrian, U. H. Differentiation and homing of IgA-secreting cells. Mucosal Immunol. 1, 96–109 (2008).

    Article  CAS  PubMed  Google Scholar 

  37. Fitzpatrick, Z. et al. Venous-plexus-associated lymphoid hubs support meningeal humoral immunity. Nature 628, 612–619 (2024).

  38. Underhill, G. H., Minges Wols, H. A., Fornek, J. L., Witte, P. L. & Kansas, G. S. IgG plasma cells display a unique spectrum of leukocyte adhesion and homing molecules. Blood 99, 2905–2912 (2002).

    Article  CAS  PubMed  Google Scholar 

  39. Pabst, O. et al. Chemokine receptor CCR9 contributes to the localization of plasma cells to the small intestine. J. Exp. Med. 199, 411–416 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Feng, N. et al. Redundant role of chemokines CCL25/TECK and CCL28/MEC in IgA+ plasmablast recruitment to the intestinal lamina propria after rotavirus infection. J. Immunol. 176, 5749–5759 (2006).

    Article  CAS  PubMed  Google Scholar 

  41. Hieshima, K. et al. CC chemokine ligands 25 and 28 play essential roles in intestinal extravasation of IgA antibody-secreting cells. J. Immunol. 173, 3668–3675 (2004).

    Article  CAS  PubMed  Google Scholar 

  42. Shimoda, M. et al. Isotype-specific selection of high affinity memory B cells in nasal-associated lymphoid tissue. J. Exp. Med. 194, 1597–1607 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Chen, H. et al. BCR selection and affinity maturation in Peyer’s patch germinal centres. Nature 582, 421–425 (2020).

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  44. Nowosad, C. R. et al. Tunable dynamics of B cell selection in gut germinal centres. Nature 588, 321–326 (2020).

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  45. Linehan, J. L. et al. Generation of Th17 cells in response to intranasal infection requires TGF-β1 from dendritic cells and IL-6 from CD301b+ dendritic cells. Proc. Natl Acad. Sci. USA 112, 12782–12787 (2015).

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  46. Lee, J. H. et al. Long-primed germinal centres with enduring affinity maturation and clonal migration. Nature 609, 998–1004 (2022).

    Article  ADS  Google Scholar 

  47. Iijima, N. & Iwasaki, A. Access of protective antiviral antibody to neuronal tissues requires CD4 T-cell help. Nature 533, 552–556 (2016).

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  48. Cornelis, R. & Shulman, Z. Upper airway and brain protection by plasma cells: a local affair. Immunity 55, 1972–1974 (2022).

    Article  CAS  PubMed  Google Scholar 

  49. Roukens, A. H. E. et al. Prolonged activation of nasal immune cell populations and development of tissue-resident SARS-CoV-2-specific CD8 T cell responses following COVID-19. Nat. Immunol. 23, 23–32 (2022).

    Article  CAS  PubMed  Google Scholar 

  50. Moseman, E. A., Blanchard, A. C., Nayak, D. & McGavern, D. B. T cell engagement of cross-presenting microglia protects the brain from a nasal virus infection. Sci. Immunol. 5, eabb1817 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Pizzolla, A. et al. Nasal-associated lymphoid tissues (NALTs) support the recall but not priming of influenza virus-specific cytotoxic T cells. Proc. Natl Acad. Sci. USA 114, 5225–5230 (2017).

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  52. Sepahi, A. & Salinas, I. The evolution of nasal immune systems in vertebrates. Mol. Immunol. 69, 131–138 (2016).

    Article  CAS  PubMed  Google Scholar 

  53. Harkema, J. R., Carey, S. A., Wagner, J. G., Dintzis, S. M. & Liggitt, D. in Comparative Anatomy and Histology (eds Treuting, P. M. & Dintzis, S. M.) 71–94, Ch. 6 (Academic, 2012).

  54. Nakashima, T. & Hamashima, Y. Local Immune System of Nasal Mucosa in Inflammation Iga Distribution and Secretory Activity. Ann. Otol. Rhinol. Laryngol. 89, 140–146 (1980).

    Article  CAS  PubMed  Google Scholar 

  55. Lundgren, A. et al. Plasmablasts in previously immunologically naïve COVID-19 patients express markers indicating mucosal homing and secrete antibodies cross-reacting with SARS-CoV-2 variants and other beta-coronaviruses. Clin. Exp. Immunol. 213, 173–189 (2023).

    Article  PubMed  PubMed Central  Google Scholar 

  56. Gruber, C. N. et al. Mapping systemic inflammation and antibody responses in multisystem inflammatory syndrome in children (MIS-C). Cell 183, 982–995 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Cisney, E. D., Fernandez, S., Hall, S. I., Krietz, G. A. & Ulrich, R. G. Examining the role of nasopharyngeal-associated lymphoreticular tissue (NALT) in mouse responses to vaccines. J. Vis. Exp. https://doi.org/10.3791/3960 (2012).

  58. Jin, J.-O. et al. Immunostimulatory agent evaluation: lymphoid tissue extraction and injection route-dependent dendritic cell activation. J. Vis. Exp. https://doi.org/10.3791/57640 (2018).

  59. Stoler-Barak, L. et al. B cell class switch recombination is regulated by DYRK1A through MSH6 phosphorylation. Nat. Commun. 14, 1462 (2023).

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  60. Kozlovski, S. et al. ICAMs are dispensable for influenza clearance and anti-viral humoral and cellular immunity. Front. Immunol. 13, 1041552 (2022).

    Article  CAS  PubMed  Google Scholar 

  61. Li, W., Germain, R. N. & Gerner, M. Y. High-dimensional cell-level analysis of tissues with Ce3D multiplex volume imaging. Nat. Protoc. 14, 1708–1733 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. Victora, G. D. et al. Germinal center dynamics revealed by multiphoton microscopy with a photoactivatable fluorescent reporter. Cell 143, 592–605 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  63. Stuart, T. et al. Comprehensive integration of single-cell data. Cell 177, 1888–1902 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  64. Jaitin, D. A. et al. Massively parallel single-cell RNA-seq for marker-free decomposition of tissues into cell types. Science 343, 776–779 (2014).

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  65. Keren-Shaul, H. et al. MARS-seq2.0: an experimental and analytical pipeline for indexed sorting combined with single-cell RNA sequencing. Nat. Protoc. 14, 1841–1862 (2019).

    Article  CAS  PubMed  Google Scholar 

  66. Kohen, R. et al. UTAP: User-friendly Transcriptome Analysis Pipeline. BMC Bioinform. 20, 154 (2019).

    Article  Google Scholar 

  67. Subramanian, A. et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl Acad. Sci. USA 102, 15545–15550 (2005).

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  68. Zhou, Y. et al. Metascape provides a biologist-oriented resource for the analysis of systems-level datasets. Nat. Commun. 10, 1523 (2019).

    Article  ADS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

Z.S. is supported by the European Research Council (ERC) grant no. 101001613, Israel Science Foundation (ISF) grant no. 1272/23 and the Morris Kahn Institute for Human Immunology. Z.S. is a member of the European Molecular Biology Organization (EMBO) Young Investigator Program. We thank M. Bemark (University of Lund) for discussions and comments about the study and D. Farfara for help in mouse head sectioning (Technion-Israel Institute of Technology).

Author information

Authors and Affiliations

  1. Department of Systems Immunology, Weizmann Institute of Science, Rehovot, Israel

    Jingjing Liu, Liat Stoler-Barak, Hadas Hezroni-Bravyi, Adi Biram & Ziv Shulman

  2. Department of Immunology and Regenerative Biology, Weizmann Institute of Science, Rehovot, Israel

    Sacha Lebon, Natalia Davidzohn & Moshe Biton

  3. Department of Life Science Core Facilities, Weizmann Institute of Science, Rehovot, Israel

    Merav Kedmi, Muriel Chemla & David Pilzer

  4. The Nancy & Stephen Grand Israel National Center for Personalized Medicine (G-INCPM), Weizmann Institute of Science, Rehovot, Israel

    Merav Kedmi, Muriel Chemla & David Pilzer

  5. Department of Veterinary Resources, Weizmann Institute of Science, Rehovot, Israel

    Marina Cohen & Ori Brenner

Authors
  1. Jingjing Liu
    View author publications

    Search author on:PubMed Google Scholar

  2. Liat Stoler-Barak
    View author publications

    Search author on:PubMed Google Scholar

  3. Hadas Hezroni-Bravyi
    View author publications

    Search author on:PubMed Google Scholar

  4. Adi Biram
    View author publications

    Search author on:PubMed Google Scholar

  5. Sacha Lebon
    View author publications

    Search author on:PubMed Google Scholar

  6. Natalia Davidzohn
    View author publications

    Search author on:PubMed Google Scholar

  7. Merav Kedmi
    View author publications

    Search author on:PubMed Google Scholar

  8. Muriel Chemla
    View author publications

    Search author on:PubMed Google Scholar

  9. David Pilzer
    View author publications

    Search author on:PubMed Google Scholar

  10. Marina Cohen
    View author publications

    Search author on:PubMed Google Scholar

  11. Ori Brenner
    View author publications

    Search author on:PubMed Google Scholar

  12. Moshe Biton
    View author publications

    Search author on:PubMed Google Scholar

  13. Ziv Shulman
    View author publications

    Search author on:PubMed Google Scholar

Contributions

J.L. designed and conducted the experiments, performed data analysis and wrote the manuscript. L.S.-B. and H.H.-B. performed RNA-seq and the bioinformatics analyses. A.B., S.L. and N.D. helped in conducting some of the experiments. M.K., M. Chemla and D.P. helped in establishing mRNA sequence libraries. M. Cohen helped in H&E staining. O.B. advised on upper airway pathology. M.B. helped in conducting sorting experiments. Z.S. conceived and supervised the study, designed experiments and wrote the manuscript. All of the authors contributed to the study and performed their work locally at the Weizmann Institute of Science.

Corresponding author

Correspondence to Ziv Shulman.

Ethics declarations

Competing interests

The authors declare no competing interests.

Peer review

Peer review information

Nature thanks the anonymous reviewers for their contribution to the peer review of this work. Peer reviewer reports are available.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data figures and tables

Extended Data Fig. 1 The NALT hosts commensal bacteria-driven chronic germinal centres.

(A) TPLSM Z-stack images of LNs derived from 8 weeks old naive AicdaCre/+ Rosa26Stop-tdTomato/+ mice (left) or AID-GFP+ (right) mice. Scale bars, 200–300 µm. AicdaCre/+ Rosa26Stop-tdTomato/+ mice, n = 3, 2 independent experiments; AID-GFP+ mice, n = 3, 3 independent experiments. (B) NALT derived from 6 week to 43 week old AicdaCre/+ Rosa26Stop-tdTomato/+ mice. Scale bars, 200 µm. One mouse in each age category was examined. (C, D) Representative plots and quantification of GC B cells derived from naive C57BL/6 SPF mice NALT and MedLN. NALT, n = 28, GC B cell frequency mean±s.e.m = 2.908 ± 0.4%; MedLN, n = 18, GC B cell frequency mean±s.e.m = 1.27 = 0.4%; data pooled from 9 independent experiments. **p = 0.0074; unpaired two-tailed Student’s t-test; (E) Representative plots and quantification of GC B cell immunoglobulin isotypes in NALT and MedLN derived from naive C57BL/6 SPF mice. IgG2b+: ***p = 0.0007, n = 12; IgG1+: p = 0.0660, n = 11; IgA+: ***p = 0.0002, n = 11; IgM+: **p = 0.0092, n = 12; 4 independent experiments, unpaired two-tailed Student’s t-test; data represent mean±s.e.m. (F) TPLSM Z-stack images of NALT and submandibular LN derived from unimmunized AicdaCre/+ Rosa26Stop-tdTomato/+ mice. Scale bars, 150 µm. n = 3, 2 independent experiments. (G) Representative plots and quantification of GC B cells in NALT and submandibular LNs from unimmunized C57BL/6 SPF mice. n = 6; 2 independent experiments. *p = 0.0237; unpaired two-tailed Student’s t-test; data represent mean±s.e.m. (H) Representative plots and quantification of GC B cell immunoglbulin isotypes. IgG2b+: p = 0.5401; IgG1+: p = 0.0809; IgA+: ***p = 0.0007; IgM+: *p = 0.0341; n = 6; 2 independent experiments; unpaired two-tailed Student’s t-test; data represent mean±s.e.m. (I) Comparison of GC B cell population between naive SPF and GF mice. SPF n = 8, GF n = 10; 2 independent experiments. NALT: ***p = 0.0006; MedLN: p = 0.7; unpaired two-tailed Student’s t-test; data represent mean ± s.e.m. Gating strategy: (C, D, G), FAS+ CD38− GC population gated from the B220+ CD45+compartment; (E, H), IgA+, IgG1+, IgG2b+ and IgM+ population gated from FAS+ CD38- B220+ compartment; (I), FAS+ CD38− GC population gated from the B220+ compartment.

Source Data

Extended Data Fig. 2 Antigen-specific B cells proliferate in the NALT.

(A) Representative plots and quantification of FITC+ B1-8hi and total B cells from NALT and blood of immunized mice that received anti-CD45-FITC antibody 10 min before the end of the experiment. CD45-FITC+ population gated from the B220+ or B1-8hi B220+ compartments. Blood n = 6, NALT n = 8; 2 independent experiments. (B) Proliferation analysis of transferred CTV labelled GFP+ B1-8hi B cells 3 days following i.n. NP-OVA + MPLA. Day 0 n = 2, day 3 n = 6; 2 independent experiments; data represent mean ± s.e.m. (C, D) Proliferation analysis of transferred GFP+ B1-8hi B cells in NALT and popliteal lymph nodes (PopLN) at day 3 (C) and day 5 (D) post i.n. NP-OVA + MPLA, 2.5 h after i.v. EdU administration. For each experiment: n = 6, 2 independent experiments, data represent mean ± s.e.m.

Extended Data Fig. 3 Additional supporting data for the scRNA-seq analysis shown in Fig. 1i.

(A, B) Two-dimensional embedding of data shown in Fig. 1i was colour-coded to represent the weight of genes associated with distinct cell clusters. Data were pooled from 5 mice. (C) Violin plots of the distribution of genes (Supplementary Data Table 1) upregulated in early memory B cells compared to naive B cells, in Ccr6+ cell clusters, either in he NALT or the MedLN. Data were pooled from 5 mice. p = 1 × 10−73. (D) Representative flow cytometry plots and quantification of CCR6+ NALT and MedLN B1-8hi B cells at day 5 following i.n. NP-OVA + MPLA. CCR6+ population gated from the GFP+ B220+ CD138− B1-8hi B cell compartment. n = 6; 3 independent experiments. ***p = 0.0002; unpaired two-tailed Student’s t-test; data represent mean ± s.e.m. (E) Supplementary data related to Fig. 1i. UMAP plots of the NALT and MedLN show colour-coded different cell cycle stages.

Source Data

Extended Data Fig. 4 Transferred antigen-specific T cells express activation markers in the NALT in response to vaccination.

(A, B) Representative TPLSM images of the NALT at day 3 post i.n. NP-OVA + MPLA. NALTs were derived from WT host mice adoptively transferred with GFP+ B1-8hi B cells and Rosa26tdTomato/+ CD4+ OT-II T cells following i.n. NP-OVA + MPLA (A), or transferred with 107 naive CFP+ B cells 1 day after i.n. NP-OVA + MPLA (B). Merged Z-stack. Scale bars, 30–150 µm. n = 3 in each experiment. (C, D) Activation marker expression on transferred OT-II T cells at day 1.5 and day 3 following i.n. NP-OVA + MPLA. NALT cells were derived from WT host mice adoptively transferred with GFP+ B1-8hi B cells and Rosa26tdTomato/+ CD4+ OT-II T cells following i.n. NP-OVA + MPLA. CD69+ and CD44high CD62L− population gated from the Rosa26tdTomato/+ CD4+ T cell compartment. C: **p = 0.0022, *p = 0.0436, ***p = 0.0002; D: **p = 0.0013, p = 0.0887, ***p = 0.0001; unimm n = 5, day 1.5, n = 7, day 3 n = 8; 2 independent experiments; unpaired two-tailed Student’s t-test; data represent mean ± s.e.m.

Source Data

Extended Data Fig. 5 B cell expansion in the SED of the NALT requires interactions with T cells.

(A, B) Representative flow cytometry plots and quantification of TFH cells in the MedLN derived from WT host mice that were adoptively transferred with H2-Ab+/+ or H2-Ab−/− Rosa26tdTomato/+ B1-8hi B cells and CD45.1+ CD4+ OT-II T cells 5 days following immunization with i.n. NP-OVA + MPLA. PD1+ CXCR5+ population gated from CD62L− CD4+ CD45.1+ OT-II T cell compartment. *p = 0.05; p = 0.60. n = 6; 2 independent experiments; unpaired two-tailed Student’s t-test; data represent mean±s.e.m. (C, D) Representative plots and quantification of GC cells in NALT (C) and MedLN (D) at day 7 following i.n. NP-OVA + MPLA with or without injection of CD4+ OT-II T cells. FAS+ CD38- GC population gated from the B220+ compartment. C, p = 0.064; D, p = 0.15. n = 6; 2 independent experiments; unpaired two-tailed Student’s t-test; data represent mean ± s.e.m. (E) Host mice were injected with anti-CD4 mAbs while transferring GFP+ B1-8hi B cells (as in Fig. 2g). Flow cytometry analysis of CD4+ T cells in the NALT at day 7 following NP-OVA i.n. boost. CD4+ T cell population gated from the CD45+ compartment. ****p = 1 × 10−5. (F-H) Experimental setting as in E. Flow cytometry quantification of B cells in NALT at day 7 following NP-OVA i.n. boost. In F, FAS+ CD38- GC population gated from B220+ compartment, p = 0.135; in G, GFP+ B1-8hi population gated from FAS+ CD38- GC compartment, *p = 0.0156; in H, IgA+ GFP+ B1-8hi B cell population gated from the B220+ compartment, **p = 0.0015; control n = 6, anti-CD4 n = 7; 2 independent experiments; unpaired two-tailed Student’s t-test; data represent mean ± s.e.m.

Source Data

Extended Data Fig. 6 Antigen-specific B cells are primarily detected in the turbinates region of i.n. vaccinated mice.

(A) TPLSM Z-stack images of GFP+ B1-8hi B cells in different tissues at day 7 post i.n. NP-OVA + MPLA boost. WT host mice were adoptively transferred with GFP+ B1-8hi B cells and Rosa26tdTomato/+ CD4+ OT-II T cells following i.n. prime and boost immunizations followed by TPLSM imaging. Scale bars, 70–200 µm. (B) TPLSM Z-stack images of the LNs after different vaccination strategies. i.n.: NP-OVA + MPLA prime and boost; i.p.+s.c.: NP-OVA+Alum i.p. prime and NP-OVA + MPLA s.c. boost; oral gavage with NP-CT. 2 independent experiment. See Supplementary Data Table 2. (C) Confocal images of the NT and TPLSM images of the NALTs after vaccination with NP-OVA in different nasal adjuvants. Numbers indicate the fraction of mice with NT B cells. 2 independent experiment. (D) TPLSM image of B1-8hi B cells in the NT. WT host mice were adoptively transferred with Blimp-1-YFP Rosa26tdTomato/+ B1-8hi B cells and CD4+ OT-II T cells following i.n. NP-OVA + MPLA prime and boost vaccination. Upper mouse heads were collected 7 days following boost. Scale bars, 50 µm. n = 5; 2 independent experiments.

Source Data

Extended Data Fig. 7 Olfactory epithelium anatomy and lymphatic structures in the NALT.

(A, C, E) Anatomy of the NT. Sagittal section of the nasal turbinates of a unimmunized mouse (8 weeks old) using H&E staining. (B, D, F) Digital slide scanner and confocal images of the nasal turbinate of naive AicdaCre/+ Rosa26Stop-tdTomato/+ Blimp-1-YFP+ mice. n = 4, 2 independent experiments. (G) TPLSM Z-stack images of the NALT derived from unimmunized Prox1-GFP+ AicdaCre/+ Rosa26Stop-tdTomato/+ mice. n = 3.

Source Data

Supplementary information

Supplementary information

Supplementary Figs. 1 and 2 and Supplementary Tables 1–3.

Reporting Summary

Peer Review File

Supplementary Video 1

TPLSM imaging of MedLN showing naive GFP+ B cells and naive Rosa26tdTomato/+ CD4+ T cells. Related to Fig. 1b. MedLN was imaged using TPLSM. Green, naive GFP+ B cells; red, naive Rosa26tdTomato/+ CD4+ T cells; blue, collagen. Most of the imaging field is occupied by B cell follicles.

Supplementary Video 2

LSFM imaging of the NALT at day 5 after i.n. NP–OVA. Related to Fig. 1d. The image is shown in volumetric visualization mode. Green, endogenous AID–GFP+ cells; red, transferred tdTomato+B1-8hi B cells.

Supplementary Video 3

LSFM imaging of the NALT at day 10 after i.n. NP–OVA. Related to Fig. 1d. The image is shown in volumetric visualization mode. Green, endogenous AID–GFP+ cells; red, transferred tdTomato+B1-8hi B cells.

Supplementary Video 4

Dynamics of the GFP+B1-8hi B cell response within the NALT at days 0 to 30 after i.n. NP–OVA. Related to Fig. 1f. NALTs were imaged using TPLSM. Green, transferred GFP+B1-8hi B cells; red, transferred tdTomato+CD4+ OT-II T cells; blue, collagen. Images appear in chronological order.

Supplementary Video 5

TPLSM image of the NALT from naive PROX1–GFP+ Aicdacre/+Rosa26Stop-tdTomato/+ mice. Related to Extended Data Fig. 7g. NALT was imaged using TPLSM. Green, PROX1–GFP+ lymphatic endothelial cells; red, Aicdacre/+Rosa26Stop-tdTomato/+ B cells; blue, collagen.

Source data

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, J., Stoler-Barak, L., Hezroni-Bravyi, H. et al. Turbinate-homing IgA-secreting cells originate in the nasal lymphoid tissues. Nature 632, 637–646 (2024). https://doi.org/10.1038/s41586-024-07729-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41586-024-07729-x

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing