Human Dectin-1 is O-glycosylated and serves as a ligand for C-type lectin receptor CLEC-2

  1. Shojiro Haji
  2. Taiki Ito
  3. Carla Guenther
  4. Miyako Nakano
  5. Takashi Shimizu
  6. Daiki Mori
  7. Yasunori Chiba
  8. Masato Tanaka
  9. Sushil K Mishra
  10. Janet A Willment
  11. Gordon D Brown
  12. Masamichi Nagae  Is a corresponding author
  13. Sho Yamasaki  Is a corresponding author
  1. Kyushu University, Japan
  2. Osaka University, Japan
  3. Hiroshima University, Japan
  4. Centre National de la Recherche Scientifique, France
  5. National Institute of Advanced Industrial Science and Technology, Japan
  6. Tokyo University of Pharmacy and Life Sciences, Japan
  7. University of Mississippi, United States
  8. University of Exeter, United Kingdom

Abstract

C-type lectin receptors (CLRs) elicit immune responses upon recognition of glycoconjugates present on pathogens and self-components. While Dectin-1 is the best-characterized CLR recognizing b-glucan on pathogens, the endogenous targets of Dectin-1 are not fully understood. Herein, we report that human Dectin-1 is a ligand for CLEC-2, another CLR expressed on platelets. Biochemical analyses revealed that Dectin-1 is a mucin-like protein as its stalk region is highly O-glycosylated. A sialylated core 1 glycan attached to the EDxxT motif of human Dectin-1, which is absent in mouse Dectin-1, provides a ligand moiety for CLEC-2. Strikingly, the expression of human Dectin-1 in mice rescued the lethality and lymphatic defect resulting from a deficiency of Podoplanin, a known CLEC-2 ligand. This finding is the first example of an innate immune receptor also functioning as a physiological ligand to regulate ontogeny upon glycosylation.

Data availability

Sequencing data have been deposited in GEO under accession code GSE196049.

The following data sets were generated

Article and author information

Author details

  1. Shojiro Haji

    Department of Molecular Immunology, Kyushu University, Fukuoka, Japan
    Competing interests
    The authors declare that no competing interests exist.
  2. Taiki Ito

    Laboratory of Molecular Immunology, Osaka University, Osaka, Japan
    Competing interests
    The authors declare that no competing interests exist.
  3. Carla Guenther

    Laboratory of Molecular Immunology, Osaka University, Osaka, Japan
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7915-1028
  4. Miyako Nakano

    Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, Japan
    Competing interests
    The authors declare that no competing interests exist.
  5. Takashi Shimizu

    Laboratory of Molecular Immunology, Osaka University, Osaka, Japan
    Competing interests
    The authors declare that no competing interests exist.
  6. Daiki Mori

    Centre National de la Recherche Scientifique, Marseille, France
    Competing interests
    The authors declare that no competing interests exist.
  7. Yasunori Chiba

    Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology, Ibaraki, Japan
    Competing interests
    The authors declare that no competing interests exist.
  8. Masato Tanaka

    Laboratory of Immune Regulation School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Hachioji, Japan
    Competing interests
    The authors declare that no competing interests exist.
  9. Sushil K Mishra

    Glycoscience Center of Research Excellence, University of Mississippi, Mississippi, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3080-9754
  10. Janet A Willment

    Medical Research Council Centre for Medical Mycology, University of Exeter, Exeter, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7040-0857
  11. Gordon D Brown

    Medical Research Council Centre for Medical Mycology, University of Exeter, Exeter, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  12. Masamichi Nagae

    Department of Molecular Immunology, Osaka University, Osaka, Japan
    For correspondence
    mnagae@biken.osaka-u.ac.jp
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5470-3807
  13. Sho Yamasaki

    Department of Molecular Immunology, Osaka University, Osaka, Japan
    For correspondence
    yamasaki@biken.osaka-u.ac.jp
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5184-6917

Funding

Japan Society for the Promotion of Science (20H00505)

  • Sho Yamasaki

Japan Society for the Promotion of Science (20K06575)

  • Masamichi Nagae

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal protocols were approved by the committee of Ethics on Animal Experiment and Research Institute for Microbial Diseases, Osaka University (Permit number: Biken-AP-R03-17-0).

Human subjects: All human subjects research was approved by the Institutional Review Board of the Research Institute for Microbial Diseases, Osaka University. Informed consent and consent to publish were obtained from all individuals donating venous blood. Consent documents and procedures were approved by the Institutional Review Board of the Research Institute for Microbial Diseases, Osaka University (Permit number 29-12).

Reviewing Editor

  1. Simon Yona, The Hebrew University of Jerusalem, Israel

Version history

  1. Received: August 28, 2022
  2. Preprint posted: September 4, 2022 (view preprint)
  3. Accepted: December 7, 2022
  4. Accepted Manuscript published: December 8, 2022 (version 1)
  5. Version of Record published: December 23, 2022 (version 2)

Copyright

© 2022, Haji et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 875
    Page views
  • 182
    Downloads
  • 1
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Shojiro Haji
  2. Taiki Ito
  3. Carla Guenther
  4. Miyako Nakano
  5. Takashi Shimizu
  6. Daiki Mori
  7. Yasunori Chiba
  8. Masato Tanaka
  9. Sushil K Mishra
  10. Janet A Willment
  11. Gordon D Brown
  12. Masamichi Nagae
  13. Sho Yamasaki
(2022)
Human Dectin-1 is O-glycosylated and serves as a ligand for C-type lectin receptor CLEC-2
eLife 11:e83037.
https://doi.org/10.7554/eLife.83037

Further reading

    1. Genetics and Genomics
    2. Immunology and Inflammation
    Roshni Roy, Pei-Lun Kuo ... Luigi Ferrucci
    Research Article Updated

    Age-associated DNA methylation in blood cells convey information on health status. However, the mechanisms that drive these changes in circulating cells and their relationships to gene regulation are unknown. We identified age-associated DNA methylation sites in six purified blood-borne immune cell types (naive B, naive CD4+ and CD8+ T cells, granulocytes, monocytes, and NK cells) collected from healthy individuals interspersed over a wide age range. Of the thousands of age-associated sites, only 350 sites were differentially methylated in the same direction in all cell types and validated in an independent longitudinal cohort. Genes close to age-associated hypomethylated sites were enriched for collagen biosynthesis and complement cascade pathways, while genes close to hypermethylated sites mapped to neuronal pathways. In silico analyses showed that in most cell types, the age-associated hypo- and hypermethylated sites were enriched for ARNT (HIF1β) and REST transcription factor (TF) motifs, respectively, which are both master regulators of hypoxia response. To conclude, despite spatial heterogeneity, there is a commonality in the putative regulatory role with respect to TF motifs and histone modifications at and around these sites. These features suggest that DNA methylation changes in healthy aging may be adaptive responses to fluctuations of oxygen availability.

    1. Immunology and Inflammation
    2. Neuroscience
    René Lemcke, Christine Egebjerg ... Birgitte R Kornum
    Research Article

    Infection with Influenza A virus (IAV) causes the well-known symptoms of the flu, including fever, loss of appetite, and excessive sleepiness. These responses, mediated by the brain, will normally disappear once the virus is cleared from the system, but a severe respiratory virus infection may cause long-lasting neurological disturbances. These include encephalitis lethargica and narcolepsy. The mechanisms behind such long lasting changes are unknown. The hypothalamus is a central regulator of the homeostatic response during a viral challenge. To gain insight into the neuronal and non-neuronal molecular changes during an IAV infection, we intranasally infected mice with an H1N1 virus and extracted the brain at different time points. Using single-nucleus RNA sequencing (snRNA-seq) of the hypothalamus, we identify transcriptional effects in all identified cell populations. The snRNA-seq data showed the most pronounced transcriptional response at 3 days past infection, with a strong downregulation of genes across all cell types. General immune processes were mainly impacted in microglia, the brain resident immune cells, where we found increased numbers of cells expressing pro-inflammatory gene networks. In addition, we found that most neuronal cell populations downregulated genes contributing to the energy homeostasis in mitochondria and protein translation in the cytosol, indicating potential reduced cellular and neuronal activity. This might be a preventive mechanism in neuronal cells to avoid intracellular viral replication and attack by phagocytosing cells. The change of microglia gene activity suggest that this is complemented by a shift in microglia activity to provide increased surveillance of their surroundings.