CoChat
Sign Up
Cited by 414Immune Response and InflammationGut microbiota and healthT-cell and B-cell Immunology

The DOI matches its Crossref record; 51 of the 52 references it cites were found in a registry or an index.

Akkermansia muciniphila phospholipid induces homeostatic immune responses

Open access
Bae, Munhyung, Cassilly, Chelsi D., Liu, Xiaoxi, Park, Sung-Moo, Tusi, Betsabeh Khoramian, Chen, Xiangjun, Kwon, Jaeyoung, Filipčík, Pavel, Bolze, Andrew S., Liu, Zehua, Vlamakis, Hera, Graham, Daniel B., Buhrlage, Sara J., Xavier, Ramnik J., Clardy, Jon·2022·Nature
DOI 10.1038/s41586-022-04985-7PMC PMC9328018
Open this paper in CoChat — free accountOpen PDF

Abstract

Abstract Multiple studies have established associations between human gut bacteria and host physiology, but determining the molecular mechanisms underlying these associations has been challenging 1–3 . Akkermansia muciniphila has been robustly associated with positive systemic effects on host metabolism, favourable outcomes to checkpoint blockade in cancer immunotherapy and homeostatic immunity 4–7 . Here we report the identification of a lipid from A. muciniphila ’s cell membrane that recapitulates the immunomodulatory activity of A. muciniphila in cell-based assays 8 . The isolated immunogen, a diacyl phosphatidylethanolamine with two branched chains (a15:0-i15:0 PE), was characterized through both spectroscopic analysis and chemical synthesis. The immunogenic activity of a15:0-i15:0 PE has a highly restricted structure–activity relationship, and its immune signalling requires an unexpected toll-like receptor TLR2–TLR1 heterodimer 9,10 . Certain features of the phospholipid’s activity are worth noting: it is significantly less potent than known natural and synthetic TLR2 agonists; it preferentially induces some inflammatory cytokines but not others; and, at low doses (1% of EC 50 ) it resets activation thresholds and responses for immune signalling. Identifying both the molecule and an equipotent synthetic analogue, its non-canonical TLR2–TLR1 signalling pathway, its immunomodulatory selectivity and its low-dose immunoregulatory effects provide a molecular mechanism for a model of A. muciniphila’ s ability to set immunological tone and its varied roles in health and disease.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1038/s41586-022-04985-7

Cites

51 of 51 references verified0 citation errors · 0 not checkable

Statistics

Figures

Fig. 1 Structural and functional analysis of A. muciniphila PE. a, Flow diagram for fractionation of A. muciniphila PE. Amounts in active fractions are shown in red. FA composition of PE fraction also shown. b, TNFα production by mBMDCs treated with A. muciniphila lipid extract fractions as measured by ELISA. The fraction indicated in red was used for structural characterization. Pam3CSK4 was used as a control agonist. Data are presented as mean values ± s.d. of technical replicates (n = 4). c, The structure of a15:0-i15:0 PE. d, The relative abundance of FAs in A. muciniphila PE. e, Dose response of TNFα production by mBMDCs treated with natural (Nat.) and synthetic (Syn.) a15:0-i15:0 PE lipids as measured by ELISA. Data are presented as mean values ± s.d. of technical replicates (n = 4). f, a15:0-i15:0 PE and complete PE (AmPE) trigger release of TNFα and IL-6 but not IL-10 or IL-12p70 from mBMDCs, as measured by flow cytometry. LPS was used as a control. Data are presented as mean values ± s.d. of technical replicates (n = 3). g, TNFα release is lost in TLR2 knockout mBMDCs but not in TLR4 knockout mBMDCs as measured by ELISA. Pam3CSK4 was used as a TLR2 control agonist, and LPS was used as a TLR4 control agonist. Data are presented as mean values ± s.d. of technical replicates (n = 4). All experiments were repeated independently at least twice with similar results. DMSO, dimethyl sulfoxide.
Fig. 1 Structural and functional analysis of A. muciniphila PE. a, Flow diagram for fractionation of A. muciniphila PE. Amounts in active fractions are shown in red. FA composition of PE fraction also shown. b, TNFα production by mBMDCs treated with A. muciniphila lipid extract fractions as measured by ELISA. The fraction indicated in red was used for structural characterization. Pam3CSK4 was used as a control agonist. Data are presented as mean values ± s.d. of technical replicates (n = 4). c, The structure of a15:0-i15:0 PE. d, The relative abundance of FAs in A. muciniphila PE. e, Dose response of TNFα production by mBMDCs treated with natural (Nat.) and synthetic (Syn.) a15:0-i15:0 PE lipids as measured by ELISA. Data are presented as mean values ± s.d. of technical replicates (n = 4). f, a15:0-i15:0 PE and complete PE (AmPE) trigger release of TNFα and IL-6 but not IL-10 or IL-12p70 from mBMDCs, as measured by flow cytometry. LPS was used as a control. Data are presented as mean values ± s.d. of technical replicates (n = 3). g, TNFα release is lost in TLR2 knockout mBMDCs but not in TLR4 knockout mBMDCs as measured by ELISA. Pam3CSK4 was used as a TLR2 control agonist, and LPS was used as a TLR4 control agonist. Data are presented as mean values ± s.d. of technical replicates (n = 4). All experiments were repeated independently at least twice with similar results. DMSO, dimethyl sulfoxide.CC BY · PubMed Central
Fig. 2 Biosynthesis and laboratory synthesis of A. muciniphila PE. a, Key genes involved in the putative biosynthetic pathway for A. muciniphila BAA-835 PE. b, Leucine or isoleucine feeding increases TNFα induction by A. muciniphila in a TLR2-dependent fashion as measured by ELISA. Pam3CSK4 and LPS were used as controls. Data are presented as mean values ± s.d. of technical replicates (n = 4). Unpaired t-test with two-tailed P value; ****P < 0.0001. c, Outline of synthetic scheme for a15:0-i15:0 PE and analogues. d, Overlay of mass spectrometric data from the natural and synthetic a15:0-i15:0. e, TNFα induction by natural and synthetic a15:0-i15:0 PE. a15:0-i15:0 PE induces production in mBMDCs, whereas n14:0-n14:0, n15:0-n15:0, n16:0-n16:0, a15:0-a15:0 and i15:0-i15:0 PE have no detectable TNFα induction, as measured by ELISA. i15:0-a15:0, the positional isomer, shows partial induction. Pam3CSK4 and LPS were used as controls. Data are presented as mean values ± s.d. of technical replicates (n = 4). All experiments were repeated independently at least twice with similar results. Ile, isoleucine; Val, valine; Leu, leucine.
Fig. 2 Biosynthesis and laboratory synthesis of A. muciniphila PE. a, Key genes involved in the putative biosynthetic pathway for A. muciniphila BAA-835 PE. b, Leucine or isoleucine feeding increases TNFα induction by A. muciniphila in a TLR2-dependent fashion as measured by ELISA. Pam3CSK4 and LPS were used as controls. Data are presented as mean values ± s.d. of technical replicates (n = 4). Unpaired t-test with two-tailed P value; ****P < 0.0001. c, Outline of synthetic scheme for a15:0-i15:0 PE and analogues. d, Overlay of mass spectrometric data from the natural and synthetic a15:0-i15:0. e, TNFα induction by natural and synthetic a15:0-i15:0 PE. a15:0-i15:0 PE induces production in mBMDCs, whereas n14:0-n14:0, n15:0-n15:0, n16:0-n16:0, a15:0-a15:0 and i15:0-i15:0 PE have no detectable TNFα induction, as measured by ELISA. i15:0-a15:0, the positional isomer, shows partial induction. Pam3CSK4 and LPS were used as controls. Data are presented as mean values ± s.d. of technical replicates (n = 4). All experiments were repeated independently at least twice with similar results. Ile, isoleucine; Val, valine; Leu, leucine.CC BY · PubMed Central
Fig. 3 TLR2–TLR1 binding model and T cell activation by a15:0-i15:0 PE. a, View of the TLR2–TLR1 complex from the Protein Data Bank (PDB ID 2z7x) with the modelled a15:0-i15:0 PE ligand in the ‘bridging’ conformation, showing the branches with C13 coloured green and C12 purple. b, An overview of the modelled TLR2–TLR1-a15:0-i15:0 PE complex in the surface representation. The dashed circle indicates the buried lipid head group. c, TLR1 and TLR2 are required for natural and synthetic A. muciniphila lipids to induce TNFα production in human monocyte-derived dendritic cells (MDDCs). The production of TNFα was measured by ELISA 18 h after adding natural or synthetic A. muciniphila lipids, Pam3CSK4, FSL-1 or LPS to cell culture media of human MDDCs following nucleofection. d, IL-23A and IL-12B induction by natural and synthetic a15:0-i15:0 PE lipids. e–g, Effects of treatment of human MDDCs with a15:0-i15:0 PE in combination with Pam3CSK4 or LPS. With long (18 h) delay times, low doses of a15:0-i15:0 PE suppress immune responses to Pam3CSK4 and moderate immune responses to LPS (e). Both effects disappear with shorter delay times (3 h in f or none in g). LPS and Pam3CSK4 were used at final concentrations of 100 ng ml−1. Data in c (n = 3), d (n = 6) and e–g (n = 4) are representative of two independent experiments, showing mean values ± s.d. P values in a were calculated by two-way analysis of variance. *P < 0.05; **P < 0.001; ****P < 0.0001; NS, not significant.
Fig. 3 TLR2–TLR1 binding model and T cell activation by a15:0-i15:0 PE. a, View of the TLR2–TLR1 complex from the Protein Data Bank (PDB ID 2z7x) with the modelled a15:0-i15:0 PE ligand in the ‘bridging’ conformation, showing the branches with C13 coloured green and C12 purple. b, An overview of the modelled TLR2–TLR1-a15:0-i15:0 PE complex in the surface representation. The dashed circle indicates the buried lipid head group. c, TLR1 and TLR2 are required for natural and synthetic A. muciniphila lipids to induce TNFα production in human monocyte-derived dendritic cells (MDDCs). The production of TNFα was measured by ELISA 18 h after adding natural or synthetic A. muciniphila lipids, Pam3CSK4, FSL-1 or LPS to cell culture media of human MDDCs following nucleofection. d, IL-23A and IL-12B induction by natural and synthetic a15:0-i15:0 PE lipids. e–g, Effects of treatment of human MDDCs with a15:0-i15:0 PE in combination with Pam3CSK4 or LPS. With long (18 h) delay times, low doses of a15:0-i15:0 PE suppress immune responses to Pam3CSK4 and moderate immune responses to LPS (e). Both effects disappear with shorter delay times (3 h in f or none in g). LPS and Pam3CSK4 were used at final concentrations of 100 ng ml−1. Data in c (n = 3), d (n = 6) and e–g (n = 4) are representative of two independent experiments, showing mean values ± s.d. P values in a were calculated by two-way analysis of variance. *P < 0.05; **P < 0.001; ****P < 0.0001; NS, not significant.CC BY · PubMed Central
Extended Data Fig. 1 Characterization of a15:0-i15:0 PE. Key COSY, HSQC, and HMBC correlations of a15:0-i15:0 PE. All experiments were repeated independently at least twice with similar results.
Extended Data Fig. 1 Characterization of a15:0-i15:0 PE. Key COSY, HSQC, and HMBC correlations of a15:0-i15:0 PE. All experiments were repeated independently at least twice with similar results.CC BY · PubMed Central
Extended Data Fig. 2 Human-cell activation by PE of A. muciniphila. a15:0-i15:0 PE induces TNFα, IL-6, IL-10, and IL-12/23p40 from human myeloid cells as measured by flow cytometry. Data are presented as mean values ± s.d. of technical replicates (n = 3). Experiments were repeated independently at least twice with similar results.
Extended Data Fig. 2 Human-cell activation by PE of A. muciniphila. a15:0-i15:0 PE induces TNFα, IL-6, IL-10, and IL-12/23p40 from human myeloid cells as measured by flow cytometry. Data are presented as mean values ± s.d. of technical replicates (n = 3). Experiments were repeated independently at least twice with similar results.CC BY · PubMed Central
Extended Data Fig. 3 Biosynthetic pathway of a15:0-i15:0 PE in A. muciniphila. Proposed biosynthesis of a15:0-i15:0 PE in A. muciniphila BAA-835.
Extended Data Fig. 3 Biosynthetic pathway of a15:0-i15:0 PE in A. muciniphila. Proposed biosynthesis of a15:0-i15:0 PE in A. muciniphila BAA-835.CC BY · PubMed Central
Extended Data Fig. 4 Biosynthetic pathways of branched-chain amino acids in A. muciniphila. De novo biosynthesis of l-leucine, l-isoleucine, and l-valine in A. muciniphila BAA-835.
Extended Data Fig. 4 Biosynthetic pathways of branched-chain amino acids in A. muciniphila. De novo biosynthesis of l-leucine, l-isoleucine, and l-valine in A. muciniphila BAA-835.CC BY · PubMed Central
Extended Data Fig. 5 TNFα activation of straight chain fatty acids. TNFα production by mBMDCs treated with single chain fatty acids (from C17:0 to C13:0) as measured by ELISA. LPS was used as a control agonist. Data are representative of at least two independent experiments and show mean values ± s.d. of technical replicates (n = 4).
Extended Data Fig. 5 TNFα activation of straight chain fatty acids. TNFα production by mBMDCs treated with single chain fatty acids (from C17:0 to C13:0) as measured by ELISA. LPS was used as a control agonist. Data are representative of at least two independent experiments and show mean values ± s.d. of technical replicates (n = 4).CC BY · PubMed Central
Extended Data Fig. 6 Heat-map of human derived monocyte activation by natural and synthetic A. muciniphila lipids. Monocytes were purified from peripheral blood by negative selection with magnetic beads. Cells were immediately placed in culture and stimulated with the indicated stimuli for 6 h. After stimulation, cells were lysed for mRNA extraction and library preparation by SmartSeq2 for RNA sequencing. Monocytes were processed from two independent donors in technical triplicate. The final concentration of bacterial lipids was 50 μg/ml. Pam3CSK4, FSL-1, and LPS at a final concentration of 100 ng/ml were used as controls.
Extended Data Fig. 6 Heat-map of human derived monocyte activation by natural and synthetic A. muciniphila lipids. Monocytes were purified from peripheral blood by negative selection with magnetic beads. Cells were immediately placed in culture and stimulated with the indicated stimuli for 6 h. After stimulation, cells were lysed for mRNA extraction and library preparation by SmartSeq2 for RNA sequencing. Monocytes were processed from two independent donors in technical triplicate. The final concentration of bacterial lipids was 50 μg/ml. Pam3CSK4, FSL-1, and LPS at a final concentration of 100 ng/ml were used as controls.CC BY · PubMed Central
Extended Data Fig. 7 Effects of treatment with synthetic A. muciniphila lipids in combination with Pam3CSK4 or LPS on human monocyte-derived dendritic cells. With long (18 h) delay times, low doses of synthetic a–c, a15:0-i15:0 PE or d–f, i15:0-a15:0 PE suppress immune responses to Pam3CSK4 and moderate immune responses to LPS. Both effects disappear with shorter delay times (3 h or none). LPS and Pam3CSK4 were used at final concentrations of 100 ng/mL. Data are presented as mean values ± s.d. of technical replicates (n = 4). Experiments were repeated independently at least twice with similar results.
Extended Data Fig. 7 Effects of treatment with synthetic A. muciniphila lipids in combination with Pam3CSK4 or LPS on human monocyte-derived dendritic cells. With long (18 h) delay times, low doses of synthetic a–c, a15:0-i15:0 PE or d–f, i15:0-a15:0 PE suppress immune responses to Pam3CSK4 and moderate immune responses to LPS. Both effects disappear with shorter delay times (3 h or none). LPS and Pam3CSK4 were used at final concentrations of 100 ng/mL. Data are presented as mean values ± s.d. of technical replicates (n = 4). Experiments were repeated independently at least twice with similar results.CC BY · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Show all 52 references

Cited by

Sorted by citations

See all 414

Related

Metadata

Authors
Bae, M., Cassilly, C. D., Liu, X., Park, S.-M., Tusi, B. K., Chen, X., Kwon, J., Filipčík, P., Bolze, A. S., Liu, Z., Vlamakis, H., Graham, D. B., Buhrlage, S. J., Xavier, R. J., & Clardy, J.
Affiliations
  • Gachon University (KR)
  • Harvard University (US)
  • Broad Institute (US)
Corresponding author
  • Xavier, Ramnik J.
  • Clardy, Jon
ORCID
Venue
Nature
Publisher
Springer Science and Business Media LLC
Published
2022-07-27
Volume
608
Issue
7921
Pages
168-173
ISSN
  • 0028-0836 (ISSN-L)
  • 1476-4687
DOI
10.1038/s41586-022-04985-7
PMC
PMC9328018
OpenAlex
W4288052200
Cited by
413
References
52
Citations by year
2026: 72 · 2025: 126 · 2024: 102 · 2023: 95 · 2022: 18
FWCI
32.66
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Digital Access to Scholarship at Harvard (DASH) (Harvard University) (acceptedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
Funders
  • Massachusetts General Hospital
  • National Institutes of Health
  • National Institute of Allergy and Infectious Diseases (R01AI172147)
  • National Institute of Diabetes and Digestive and Kidney Diseases (RC2DK114784, P30DK043351, R01DK131742, R01DK127171)
  • National Center for Complementary and Integrative Health (F32AT010415, R01AT009708)
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gut microbiota and health
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
Keywords
Akkermansia muciniphila, immunomodulation, Toll-like receptor signaling, structure-activity relationship, gut microbiota, immune homeostasis, inflammatory cytokines, synthetic analogues
MeSH terms
Akkermansia, Cell Membrane, Homeostasis, Humans, Immunity, Phosphatidylethanolamines, Structure-Activity Relationship, Cytokines, Inflammation Mediators, Toll-Like Receptor 1, Toll-Like Receptor 2

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review