1. Immunology and Inflammation
  2. Microbiology and Infectious Disease

Pathogen infection and cholesterol deficiency activate the C. elegans p38 immune pathway through a TIR-1/SARM1 phase transition

  1. Nicholas D Peterson
  2. Janneke D Icso
  3. J. Elizabeth Salisbury
  4. Tomás Rodríguez
  5. Paul R Thompson  Is a corresponding author
  6. Read Pukkila-Worley  Is a corresponding author
  1. University of Massachusetts Chan Medical School, United States
  2. University of Massachusetts Medical School, United States
https://doi.org/10.7554/eLife.74206
Share this article
Cite this article
  1. Nicholas D Peterson
  2. Janneke D Icso
  3. J. Elizabeth Salisbury
  4. Tomás Rodríguez
  5. Paul R Thompson
  6. Read Pukkila-Worley
(2022)
Pathogen infection and cholesterol deficiency activate the C. elegans p38 immune pathway through a TIR-1/SARM1 phase transition
eLife 11:e74206.
https://doi.org/10.7554/eLife.74206
  2. Download BibTeX
  3. Download .RIS

Abstract

Intracellular signaling regulators can be concentrated into membrane-free, higher-ordered protein assemblies to initiate protective responses during stress - a process known as phase transition. Here, we show that a phase transition of the Caenorhabditis elegans Toll/interleukin-1 receptor domain protein (TIR-1), an NAD+ glycohydrolase homologous to mammalian sterile alpha and TIR motif-containing 1 (SARM1), underlies p38 PMK-1 immune pathway activation in C. elegans intestinal epithelial cells. Through visualization of fluorescently labeled TIR-1/SARM1 protein, we demonstrate that physiologic stresses, both pathogen and non-pathogen, induce multimerization of TIR-1/SARM1 into visible puncta within intestinal epithelial cells. In vitro enzyme kinetic analyses revealed that, like mammalian SARM1, the NAD+ glycohydrolase activity of C. elegans TIR-1 is dramatically potentiated by protein oligomerization and a phase transition. Accordingly, C. elegans with genetic mutations that specifically block either multimerization or the NAD+ glycohydrolase activity of TIR-1/SARM1 fail to induce p38 PMK phosphorylation, are unable to increase immune effector expression, and are dramatically susceptible to bacterial infection. Finally, we demonstrate that a loss-of-function mutation in nhr-8, which alters cholesterol metabolism and is used to study conditions of sterol deficiency, causes TIR-1/SARM1 to oligomerize into puncta in intestinal epithelial cells. Cholesterol scarcity increases p38 PMK-1 phosphorylation, primes immune effector induction in a manner that requires TIR-1/SARM1 oligomerization and its intrinsic NAD+ glycohydrolase activity, and reduces pathogen accumulation in the intestine during a subsequent infection. These data reveal a new adaptive response that allows a metazoan host to anticipate pathogen threats during cholesterol deprivation, a time of relative susceptibility to infection. Thus, a phase transition of TIR-1/SARM1 as a prerequisite for its NAD+ glycohydrolase activity is strongly conserved across millions of years of evolution and is essential for diverse physiological processes in multiple cell types.

Data availability

The mRNA-seq datasets are available from the NCBI Gene Expression Omnibus using the accession numbers GSE178572 and GSE190585.Source data files are provided for all figures.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Nicholas D Peterson

    Program in Innate Immunity, University of Massachusetts Chan Medical School, Worcester, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4157-8119

  2. Janneke D Icso

    Program in Chemical Biology, University of Massachusetts Medical School, Worcester, United States
    Competing interests
    The authors declare that no competing interests exist.

  3. J. Elizabeth Salisbury

    Program in Innate Immunity, University of Massachusetts Chan Medical School, Worcester, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. Tomás Rodríguez

    RNA Therapeutics Institute, University of Massachusetts Chan Medical School, Worcester, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8724-5427

  5. Paul R Thompson

    Department of Biochemistry and Pharmacology, University of Massachusetts Medical School, Worcester, United States
    For correspondence
    Paul.Thompson@umassmed.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1621-3372

  6. Read Pukkila-Worley

    Program in Innate Immunity, University of Massachusetts Chan Medical School, Worcester, United States
    For correspondence
    read.pukkila-worley@umassmed.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5340-8294

Funding

National Institute of Allergy and Infectious Diseases (R01 AI130289)

  • Read Pukkila-Worley

National Institute of Allergy and Infectious Diseases (R01 AI159159)

  • Read Pukkila-Worley

National Institute of Allergy and Infectious Diseases (R21 AI163430)

  • Read Pukkila-Worley

Kenneth Rainin Foundation (Innovator Award)

  • Read Pukkila-Worley

National Institute of Allergy and Infectious Diseases (F30 AI150127)

  • Nicholas D Peterson

National Institute of Allergy and Infectious Diseases (T32 AI132152)

  • Nicholas D Peterson
  • Janneke D Icso

National Institute of General Medical Sciences (T32 GM107000)

  • Nicholas D Peterson

National Institute of Neurological Disorders and Stroke (F31 NS122423)

  • Janneke D Icso

National Institute of General Medical Sciences (R35 GM118112)

  • Paul R Thompson

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

Copyright

© 2022, Peterson 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

  • 3,960
    views
  • 526
    downloads
  • 46
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

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. Nicholas D Peterson
  2. Janneke D Icso
  3. J. Elizabeth Salisbury
  4. Tomás Rodríguez
  5. Paul R Thompson
  6. Read Pukkila-Worley
(2022)
Pathogen infection and cholesterol deficiency activate the C. elegans p38 immune pathway through a TIR-1/SARM1 phase transition
eLife 11:e74206.
https://doi.org/10.7554/eLife.74206

Share this article

https://doi.org/10.7554/eLife.74206

Categories and tags

Research organism

Further reading

    1. Immunology and Inflammation
    2. Medicine

    JAK inhibition decreases the autoimmune burden in Down syndrome

    Angela L Rachubinski, Elizabeth Wallace ... Joaquín M Espinosa
    Research Article

    Background:

    Individuals with Down syndrome (DS), the genetic condition caused by trisomy 21 (T21), display clear signs of immune dysregulation, including high rates of autoimmunity and severe complications from infections. Although it is well established that T21 causes increased interferon responses and JAK/STAT signaling, elevated autoantibodies, global immune remodeling, and hypercytokinemia, the interplay between these processes, the clinical manifestations of DS, and potential therapeutic interventions remain ill defined.

    Methods:

    We report a comprehensive analysis of immune dysregulation at the clinical, cellular, and molecular level in hundreds of individuals with DS, including autoantibody profiling, cytokine analysis, and deep immune mapping. We also report the interim analysis of a Phase II clinical trial investigating the safety and efficacy of the JAK inhibitor tofacitinib through multiple clinical and molecular endpoints.

    Results:

    We demonstrate multi-organ autoimmunity of pediatric onset concurrent with unexpected autoantibody-phenotype associations in DS. Importantly, constitutive immune remodeling and hypercytokinemia occur from an early age prior to autoimmune diagnoses or autoantibody production. Analysis of the first 10 participants to complete 16 weeks of tofacitinib treatment shows a good safety profile and no serious adverse events. Treatment reduced skin pathology in alopecia areata, psoriasis, and atopic dermatitis, while decreasing interferon scores, cytokine scores, and levels of pathogenic autoantibodies without overt immune suppression.

    Conclusions:

    JAK inhibition is a valid strategy to treat autoimmune conditions in DS. Additional research is needed to define the effects of JAK inhibition on the broader developmental and clinical hallmarks of DS.

    Funding:

    NIAMS, Global Down Syndrome Foundation.

    Clinical trial number:

    NCT04246372.

    1. Immunology and Inflammation

    Downregulation of semaphorin 4A in keratinocytes reflects the features of non-lesional psoriasis

    Miki Kume, Hanako Koguchi-Yoshioka ... Rei Watanabe
    Research Article

    Psoriasis is a multifactorial disorder mediated by IL-17-producing T cells, involving immune cells and skin-constituting cells. Semaphorin 4A (Sema4A), an immune semaphorin, is known to take part in T helper type 1/17 differentiation and activation. However, Sema4A is also crucial for maintaining peripheral tissue homeostasis and its involvement in skin remains unknown. Here, we revealed that while Sema4A expression was pronounced in psoriatic blood lymphocytes and monocytes, it was downregulated in the keratinocytes of both psoriatic lesions and non-lesions compared to controls. Imiquimod application induced more severe dermatitis in Sema4A knockout (KO) mice compared to wild-type (WT) mice. The naïve skin of Sema4A KO mice showed increased T cell infiltration and IL-17A expression along with thicker epidermis and distinct cytokeratin expression compared to WT mice, which are hallmarks of psoriatic non-lesions. Analysis of bone marrow chimeric mice suggested that Sema4A expression in keratinocytes plays a regulatory role in imiquimod-induced dermatitis. The epidermis of psoriatic non-lesion and Sema4A KO mice demonstrated mTOR complex 1 upregulation, and the application of mTOR inhibitors reversed the skewed expression of cytokeratins in Sema4A KO mice. Conclusively, Sema4A-mediated signaling cascades can be triggers for psoriasis and targets in the treatment and prevention of psoriasis.

    1. Immunology and Inflammation

    TCR transgenic clone selection guided by immune receptor analysis and single-cell RNA expression of polyclonal responders

    Nincy Debeuf, Sahine Lameire ... Bart N Lambrecht
    Research Article

    Since the precursor frequency of naive T cells is extremely low, investigating the early steps of antigen-specific T cell activation is challenging. To overcome this detection problem, adoptive transfer of a cohort of T cells purified from T cell receptor (TCR) transgenic donors has been extensively used but is not readily available for emerging pathogens. Constructing TCR transgenic mice from T cell hybridomas is a labor-intensive and sometimes erratic process, since the best clones are selected based on antigen-induced CD69 upregulation or IL-2 production in vitro, and TCR chains are polymerase chain reaction (PCR)-cloned into expression vectors. Here, we exploited the rapid advances in single-cell sequencing and TCR repertoire analysis to select the best clones without hybridoma selection, and generated CORSET8 mice (CORona Spike Epitope specific CD8 T cell), carrying a TCR specific for the Spike protein of SARS-CoV-2. Implementing newly created DALI software for TCR repertoire analysis in single-cell analysis enabled the rapid selection of the ideal responder CD8 T cell clone, based on antigen reactivity, proliferation, and immunophenotype in vivo. Identified TCR sequences were inserted as synthetic DNA into an expression vector and transgenic CORSET8 donor mice were created. After immunization with Spike/CpG-motifs, mRNA vaccination or SARS-CoV-2 infection, CORSET8 T cells strongly proliferated and showed signs of T cell activation. Thus, a combination of TCR repertoire analysis and scRNA immunophenotyping allowed rapid selection of antigen-specific TCR sequences that can be used to generate TCR transgenic mice.