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Cited by 856Multiple Sclerosis Research StudiesNeuroinflammation and Neurodegeneration MechanismsT-cell and B-cell Immunology

The DOI matches its Crossref record; 49 of the 51 references it cites were found in a registry or an index; 26 of the 47 sources its claims cite were checked for support.

Differential roles of microglia and monocytes in the inflamed central nervous system

Open access
Yamasaki, Ryo, Lu, Haiyan, Butovsky, Oleg, Ohno, Nobuhiko, Rietsch, Anna M., Cialic, Ron, Wu, Pauline M., Doykan, Camille E., Lin, Jessica, Cotleur, Anne C., Kidd, Grahame, Zorlu, Musab M., Sun, Nathan, Hu, Weiwei, Liu, LiPing, Lee, Jar-Chi, Taylor, Sarah E., Uehlein, Lindsey, Dixon, Debra, Gu, Jinyu, Floruta, Crina M., Zhu, Min, Charo, Israel F., Weiner, Howard L., Ransohoff, Richard M.·2014·Journal of Experimental Medicine
DOI 10.1084/jem.20132477PMC PMC4113947
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Abstract

In the human disorder multiple sclerosis (MS) and in the model experimental autoimmune encephalomyelitis (EAE), macrophages predominate in demyelinated areas and their numbers correlate to tissue damage. Macrophages may be derived from infiltrating monocytes or resident microglia, yet are indistinguishable by light microscopy and surface phenotype. It is axiomatic that T cell–mediated macrophage activation is critical for inflammatory demyelination in EAE, yet the precise details by which tissue injury takes place remain poorly understood. In the present study, we addressed the cellular basis of autoimmune demyelination by discriminating microglial versus monocyte origins of effector macrophages. Using serial block-face scanning electron microscopy (SBF-SEM), we show that monocyte-derived macrophages associate with nodes of Ranvier and initiate demyelination, whereas microglia appear to clear debris. Gene expression profiles confirm that monocyte-derived macrophages are highly phagocytic and inflammatory, whereas those arising from microglia demonstrate an unexpected signature of globally suppressed cellular metabolism at disease onset. Distinguishing tissue-resident macrophages from infiltrating monocytes will point toward new strategies to treat disease and promote repair in diverse inflammatory pathologies in varied organs.

Overview

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Written from the paper’s full text.

TL;DR

Using dual reporter mice and serial block-face scanning electron microscopy, this study shows that monocyte-derived macrophages (MDMs) initiate demyelination at EAE onset, often at nodes of Ranvier, while microglia-derived macrophages (MiDMs) clear debris and show a suppressed metabolic gene signature. Expression profiling further indicates that MDMs and MiDMs have distinct effector programs despite sharing the same inflammatory CNS environment.

Key findings

  • At EAE onset, MDMs and MiDMs were present in equal numbers; MiDM expansion began at peak, and at recovery MiDMs returned near preonset numbers while MDM frequency continued to decline.
  • Most axoglial units (55/75; 73%) contacted both MDMs and MiDMs, but where only one myeloid type was present (20/75; 27%), nearly all were MDMs (18/20); 2–3 MDMs attached to each of 18/20 (90%) such units.
  • MDMs contacted nodes of Ranvier in 7/75 (9%) axoglial units and showed invasive/demyelinating behavior; no MiDMs contacted nodes of Ranvier. In CCR2-deficient mice, demyelination was significantly reduced at EAE onset.
  • MDM-enriched genes more often encoded effector functions than MiDM-enriched genes (18/28; 64.3% vs. 18/48; 37.5%), and MiDMs showed down-regulation of metabolic, activation, phagocytosis, and reactive oxygen species–related functions at onset.

Methods

  • EAE was induced with MOG35–55 in Ccr2rfp/+::Cx3cr1gfp/+ and Ccr2rfp/rfp::Cx3cr1gfp/+ mice; CNS mononuclear cells were analyzed by flow cytometry and sorted as F4/80+RFP+ MDMs or F4/80+GFP+ MiDMs.
  • Confocal 3D morphometry and serial block-face scanning electron microscopy (SBF-SEM) were used to distinguish MDMs from MiDMs and quantify their relationships to axoglial units, nodes of Ranvier, myelin debris, and demyelination.
  • nCounter digital multiplexed gene expression of 179 inflammation-related genes was performed on naive microglia/monocytes and sorted MiDMs/MDMs across EAE onset, peak, and recovery, with hierarchical clustering, k-means clustering, and Ingenuity IPA pathway analysis.

Limitations

  • The paper states that the study focused on a single pathogenically relevant time point, EAE onset, so it could not decipher the phenotypic fate of infiltrated monocytes.
  • The authors state that conclusions should not be generalized beyond the present disease paradigm, noting that other models such as spinal cord contusion include Ly6Clow/CX3CR1gfp monocytes that are highly pathogenic.
  • Evident from the text: the ultrastructural criteria used to distinguish MDMs and MiDMs may not be universally present in other pathological circumstances, and the findings are from mouse EAE rather than direct human MS tissue.

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Registry

Crossref · 10.1084/jem.20132477

Cites

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Figures

Figure 1. MDMs and MiDMs exhibit different time courses of accumulation in the CNS of mice with EAE and morphological characteristics can distinguish them. (A) Immunohistochemistry shows expression of CD11b for red RFP+ MDMs and green GFP+ MiDMs in the spinal cords of Cx3cr1gfp/+::Ccr2rfp/+ mice at EAE onset. Bars: 25 µm. We studied 6 mice at EAE onset from 3 EAE inductions. In each EAE induction, 8–10 mice were used and 2 mice were selected from each induction. (B) Flow cytometric analysis of CCR2-RFP+ and CX3CR1-GFP+ populations in cells gated for F4/80 expression (top); CD45 expression of F4/80+RFP+ MDMs and F4/80+GFP+ MiDMs populations (middle); and MDMs and MiDMs numbers at EAE onset, peak, and recovery (bottom). We studied 3 mice for naive groups; 12 for onset; 15 for peak; 13 for recovery from 5 EAE inductions. For each induction, 8–10 mice were used and 2–3 mice were selected at each time point (onset, peak, and recovery) for analysis. (C) Confocal microscopy assessment of myeloid cell morphology in lumbar spinal cord from mice at EAE onset. We studied 54 MDMs and 51 MiDMs of 5 EAE onset mice from 3 EAE inductions for (C–E); 2 sections/mouse; 4–6 cells/section; 8–12 cells /mouse. In each EAE induction, 8–10 mice were induced and 1–2 EAE onset mice were selected from each experiment. Bars, 25 µm. (D) Cell volumes of 500 µm3; surface areas of 1,000 µm2; primary process numbers ≤3 or ≥5; solidity3D of 0.4; and Formfactor3D of 0.3 discriminate between MDMs and MiDMs. (E) Model plot of cell volume against primary process number to distinguish MDMs (red symbols and pink area) from MiDMs (green symbols and green area).
Figure 1. MDMs and MiDMs exhibit different time courses of accumulation in the CNS of mice with EAE and morphological characteristics can distinguish them. (A) Immunohistochemistry shows expression of CD11b for red RFP+ MDMs and green GFP+ MiDMs in the spinal cords of Cx3cr1gfp/+::Ccr2rfp/+ mice at EAE onset. Bars: 25 µm. We studied 6 mice at EAE onset from 3 EAE inductions. In each EAE induction, 8–10 mice were used and 2 mice were selected from each induction. (B) Flow cytometric analysis of CCR2-RFP+ and CX3CR1-GFP+ populations in cells gated for F4/80 expression (top); CD45 expression of F4/80+RFP+ MDMs and F4/80+GFP+ MiDMs populations (middle); and MDMs and MiDMs numbers at EAE onset, peak, and recovery (bottom). We studied 3 mice for naive groups; 12 for onset; 15 for peak; 13 for recovery from 5 EAE inductions. For each induction, 8–10 mice were used and 2–3 mice were selected at each time point (onset, peak, and recovery) for analysis. (C) Confocal microscopy assessment of myeloid cell morphology in lumbar spinal cord from mice at EAE onset. We studied 54 MDMs and 51 MiDMs of 5 EAE onset mice from 3 EAE inductions for (C–E); 2 sections/mouse; 4–6 cells/section; 8–12 cells /mouse. In each EAE induction, 8–10 mice were induced and 1–2 EAE onset mice were selected from each experiment. Bars, 25 µm. (D) Cell volumes of 500 µm3; surface areas of 1,000 µm2; primary process numbers ≤3 or ≥5; solidity3D of 0.4; and Formfactor3D of 0.3 discriminate between MDMs and MiDMs. (E) Model plot of cell volume against primary process number to distinguish MDMs (red symbols and pink area) from MiDMs (green symbols and green area).CC BY-NC-SA · PubMed Central
Figure 2. SBF-SEM shows MDMs initiating demyelination at EAE onset. Quantitation of MiDMs and MDMs interacting with axoglial units in SBF-SEM images of CNS at EAE onset. Intact (69%) and demyelinated (76%) segments interacted with MDMs and MiDMs. Red and pink, MDMs; green and light green, MiDMs; yellow, both MDMs and MiDMs. We studied 29 intact axon segments, 46 demyelinated axon segments, 86 MiDMs, and 169 MDMs in 14 lesions of 7 EAE onset mice from 3 EAE inductions as follows: 8–10 mice were immunized at each experiment and 2–3 onset mice were selected from each induction.
Figure 2. SBF-SEM shows MDMs initiating demyelination at EAE onset. Quantitation of MiDMs and MDMs interacting with axoglial units in SBF-SEM images of CNS at EAE onset. Intact (69%) and demyelinated (76%) segments interacted with MDMs and MiDMs. Red and pink, MDMs; green and light green, MiDMs; yellow, both MDMs and MiDMs. We studied 29 intact axon segments, 46 demyelinated axon segments, 86 MiDMs, and 169 MDMs in 14 lesions of 7 EAE onset mice from 3 EAE inductions as follows: 8–10 mice were immunized at each experiment and 2–3 onset mice were selected from each induction.CC BY-NC-SA · PubMed Central
Figure 3. SBF-SEM shows example of MDM-initiating demyelination at EAE onset. (A) Representative MDMs encircles the axoglial unit. A myelin ovoid within an intracellular phagolysosome shows physical continuity with myelin remaining attached to an axoglial unit which is undergoing active demyelination. In serial images, disrupted myelin shows continuity from outside to inside the MDM. (B) Rotated view from B demonstrating MDM-extensive attachment to axoglial unit and MiDM nearby with limited attachment to axon. A, axon; m, myelin; c, cytosol; n, nucleus. Red, MDM cytosol; green, MiDM cytosol; yellow: nuclei; blue, myelin and myelin debris; gray, axoplasm; red line, MDM plasma membrane. We studied 14 lesions from 7 EAE onset mice from 3 EAE inductions as follows: 8–10 mice were immunized at each experiment and 2–3 EAE onset mice were selected from each induction. Bar, 2 µm.
Figure 3. SBF-SEM shows example of MDM-initiating demyelination at EAE onset. (A) Representative MDMs encircles the axoglial unit. A myelin ovoid within an intracellular phagolysosome shows physical continuity with myelin remaining attached to an axoglial unit which is undergoing active demyelination. In serial images, disrupted myelin shows continuity from outside to inside the MDM. (B) Rotated view from B demonstrating MDM-extensive attachment to axoglial unit and MiDM nearby with limited attachment to axon. A, axon; m, myelin; c, cytosol; n, nucleus. Red, MDM cytosol; green, MiDM cytosol; yellow: nuclei; blue, myelin and myelin debris; gray, axoplasm; red line, MDM plasma membrane. We studied 14 lesions from 7 EAE onset mice from 3 EAE inductions as follows: 8–10 mice were immunized at each experiment and 2–3 EAE onset mice were selected from each induction. Bar, 2 µm.CC BY-NC-SA · PubMed Central
Figure 4. MDMs surrounded, apposed, and invaded nodes of Ranvier at EAE onset. (A) SBF-SEM images and 3D reconstruction of SBF-SEM images of MDMs with a node of Ranvier. White and black arrow: microvillus. (B) SBF-SEM images and 3D construction of intratubal MDMs with demyelinated axon and node of Ranvier. (C) SBF-SEM images and 3D reconstruction of an MDM with intracellular myelin debris apposed to a node of Ranvier. Red, MDM cytosol; yellow, nucleus; blue, myelin; gray, axoplasm. M, myelin; c, cytosol. red line, MDM plasma membrane. We studied 14 lesions from 7 EAE onset mice collected as follows: 8–10 mice were immunized at each induction and 2–3 EAE onset mice were selected from each immunization. Bar, 2 µm.
Figure 4. MDMs surrounded, apposed, and invaded nodes of Ranvier at EAE onset. (A) SBF-SEM images and 3D reconstruction of SBF-SEM images of MDMs with a node of Ranvier. White and black arrow: microvillus. (B) SBF-SEM images and 3D construction of intratubal MDMs with demyelinated axon and node of Ranvier. (C) SBF-SEM images and 3D reconstruction of an MDM with intracellular myelin debris apposed to a node of Ranvier. Red, MDM cytosol; yellow, nucleus; blue, myelin; gray, axoplasm. M, myelin; c, cytosol. red line, MDM plasma membrane. We studied 14 lesions from 7 EAE onset mice collected as follows: 8–10 mice were immunized at each induction and 2–3 EAE onset mice were selected from each immunization. Bar, 2 µm.CC BY-NC-SA · PubMed Central
Figure 5. Nodal pathology without demyelination at EAE onset in Ccr2rfp/rfp::Cx3cr1gfp/+ mice. (A) Magnitude of weight loss in Ccr2rfp/+::Cx3cr1gfp/+ and Ccr2rfp/rfp::Cx3cr1gfp/+ mice at preonset and onset stages of EAE. Clinical score in Ccr2rfp/+::Cx3cr1gfp/+ and Ccr2rfp/rfp::Cx3cr1gfp/+ mice at EAE onset stage. (B) Days at disease preonset and onset stages of EAE. We studied 28 Ccr2rfp/+::Cx3cr1gfp/+ mice and 26 Ccr2rfp/rfp::Cx3cr1gfp/+ mice for A and B. Data were collected from 12 EAE inductions in Ccr2rfp/+::Cx3cr1gfp/+ mice and 19 EAE inductions in Ccr2rfp/rfp::Cx3cr1gfp/+ mice as follows: 8–10 mice were immunized at each induction and 1–3 EAE recovery mice were selected from each immunization. **, P < 0.01; ***, P < 0.001. (C) SBF-SEM imaging of MDMs with nodes of Ranvier phagocytosis in Ccr2rfp/+::Cx3cr1gfp/+ mice at EAE preonset. Pink, MDM cytosol; red arrow, myelin inclusion of MDM connecting to the node of Ranvier. We studied 3 tissues from 3 Ccr2rfp/+::Cx3cr1gfp/+ EAE mice at preonset stage from 3 EAE inductions: 8–10 mice were immunized at each experiment and one EAE preonset mouse was selected from each induction. Bar, 2 µm. (D) SBF-SEM of disrupted nodes (black arrows) in preonset spinal cord tissues of Ccr2rfp/rfp::Cx3cr1gfp/+ mice. Bar, 2 µm. (E) SBF-SEM of neutrophil is with myelin phagocytosis from internode at preonset stage of Ccr2rfp/rfp::Cx3cr1gfp/+ mouse. Blue, neutrophil cytosol. For D–E, we studied three tissues from three Ccr2rfp/rfp::Cx3cr1gfp/+ EAE mice at preonset stage from 3 EAE inductions: 8–10 mice were immunized at each experiment and one EAE preonset mouse was selected from each induction. Bar: 2 µm. (F) Histochemical staining and with aurohalophosphate complexes (black gold staining) and quantification of demyelinated area of Ccr2rfp/+::Cx3cr1gfp/+ mice and Ccr2rfp/+::Cx3cr1gfp/+ mice. We studied 5 naive Ccr2rfp/+::Cx3cr1gfp/+ mice, 5 naive Ccr2rfp/rfp::Cx3cr1gfp/+ mice, 5 onset Ccr2rfp/+::Cx3cr1gfp/+ mice, and 5 onset Ccr2rfp/rfp::Cx3cr1gfp/+ mice from 3 EAE inductions as follows: 8–10 mice were immunized at each experiment and 1–2 onset mice were selected from each induction. **, P < 0.01. Bar: 250 µm.
Figure 5. Nodal pathology without demyelination at EAE onset in Ccr2rfp/rfp::Cx3cr1gfp/+ mice. (A) Magnitude of weight loss in Ccr2rfp/+::Cx3cr1gfp/+ and Ccr2rfp/rfp::Cx3cr1gfp/+ mice at preonset and onset stages of EAE. Clinical score in Ccr2rfp/+::Cx3cr1gfp/+ and Ccr2rfp/rfp::Cx3cr1gfp/+ mice at EAE onset stage. (B) Days at disease preonset and onset stages of EAE. We studied 28 Ccr2rfp/+::Cx3cr1gfp/+ mice and 26 Ccr2rfp/rfp::Cx3cr1gfp/+ mice for A and B. Data were collected from 12 EAE inductions in Ccr2rfp/+::Cx3cr1gfp/+ mice and 19 EAE inductions in Ccr2rfp/rfp::Cx3cr1gfp/+ mice as follows: 8–10 mice were immunized at each induction and 1–3 EAE recovery mice were selected from each immunization. **, P < 0.01; ***, P < 0.001. (C) SBF-SEM imaging of MDMs with nodes of Ranvier phagocytosis in Ccr2rfp/+::Cx3cr1gfp/+ mice at EAE preonset. Pink, MDM cytosol; red arrow, myelin inclusion of MDM connecting to the node of Ranvier. We studied 3 tissues from 3 Ccr2rfp/+::Cx3cr1gfp/+ EAE mice at preonset stage from 3 EAE inductions: 8–10 mice were immunized at each experiment and one EAE preonset mouse was selected from each induction. Bar, 2 µm. (D) SBF-SEM of disrupted nodes (black arrows) in preonset spinal cord tissues of Ccr2rfp/rfp::Cx3cr1gfp/+ mice. Bar, 2 µm. (E) SBF-SEM of neutrophil is with myelin phagocytosis from internode at preonset stage of Ccr2rfp/rfp::Cx3cr1gfp/+ mouse. Blue, neutrophil cytosol. For D–E, we studied three tissues from three Ccr2rfp/rfp::Cx3cr1gfp/+ EAE mice at preonset stage from 3 EAE inductions: 8–10 mice were immunized at each experiment and one EAE preonset mouse was selected from each induction. Bar: 2 µm. (F) Histochemical staining and with aurohalophosphate complexes (black gold staining) and quantification of demyelinated area of Ccr2rfp/+::Cx3cr1gfp/+ mice and Ccr2rfp/+::Cx3cr1gfp/+ mice. We studied 5 naive Ccr2rfp/+::Cx3cr1gfp/+ mice, 5 naive Ccr2rfp/rfp::Cx3cr1gfp/+ mice, 5 onset Ccr2rfp/+::Cx3cr1gfp/+ mice, and 5 onset Ccr2rfp/rfp::Cx3cr1gfp/+ mice from 3 EAE inductions as follows: 8–10 mice were immunized at each experiment and 1–2 onset mice were selected from each induction. **, P < 0.01. Bar: 250 µm.CC BY-NC-SA · PubMed Central
Figure 6. Inflammatory signature in MiDMs versus MDMs in the CNS of Cx3cr1gfp/+::Ccr2rfp/+ mice with EAE. (A) Quantitative nCounter expression profiling of 179 inflammation related genes was performed in CNS-derived GFP+ microglia and RFP+ recruited monocytes from naive and EAE mice at onset, peak and recovery stages. Each row of the heat map represents an individual gene and each column an individual group from pool of 5 mice at each time point. The relative abundance of transcripts is indicated by a color (red, higher; green, median; blue, low). For A–H, we studied five mice in each time point (onset, peak, recovery) from 3 EAE inductions; 8–10 mice were immunized in each induction. (B) Heat map of differentially expressed microglia and monocyte genes. (C) Enriched monocyte genes as compared with resident microglia. Bars represent fold changes of gene expression across naive and all disease stages versus resident microglia. (D) Enriched microglia genes as compared with recruited monocytes. Bars represent fold changes of gene expression across naive and all disease stages versus recruited monocytes. (E–H) K-means clustering of inflammation genes in resident microglia and recruited monocytes. K-means clustering was used to generate 5 disease stage–related clusters in MiDMs. Heat map (E) and expression profile (F) of inflammation genes in MiDMs are shown by generated clusters. MDM expression matrix overlaid on microglial based clusters shows (G) heat map and (H) expression profile.
Figure 6. Inflammatory signature in MiDMs versus MDMs in the CNS of Cx3cr1gfp/+::Ccr2rfp/+ mice with EAE. (A) Quantitative nCounter expression profiling of 179 inflammation related genes was performed in CNS-derived GFP+ microglia and RFP+ recruited monocytes from naive and EAE mice at onset, peak and recovery stages. Each row of the heat map represents an individual gene and each column an individual group from pool of 5 mice at each time point. The relative abundance of transcripts is indicated by a color (red, higher; green, median; blue, low). For A–H, we studied five mice in each time point (onset, peak, recovery) from 3 EAE inductions; 8–10 mice were immunized in each induction. (B) Heat map of differentially expressed microglia and monocyte genes. (C) Enriched monocyte genes as compared with resident microglia. Bars represent fold changes of gene expression across naive and all disease stages versus resident microglia. (D) Enriched microglia genes as compared with recruited monocytes. Bars represent fold changes of gene expression across naive and all disease stages versus recruited monocytes. (E–H) K-means clustering of inflammation genes in resident microglia and recruited monocytes. K-means clustering was used to generate 5 disease stage–related clusters in MiDMs. Heat map (E) and expression profile (F) of inflammation genes in MiDMs are shown by generated clusters. MDM expression matrix overlaid on microglial based clusters shows (G) heat map and (H) expression profile.CC BY-NC-SA · PubMed Central
Figure 7. Affected functions in MiDMs and MDMs at EAE onset. nCounter inflammatory gene expression data were uploaded to IPA. Genes with fold change (EAE onset vs. Naive) ≥1.5 or ≤−1.5 were included in downstream effects analysis. (A) MDMs up-regulated functions, sorted by activation z-score. (B) MiDMs up-regulated (left) and down-regulated (right) functions, sorted by activation z score. The bias term indicates imbalanced numbers of up- and down-regulated genes associated with a distal function requiring significance at the P < 0.01 level. We studied pooled samples from 5 mice in each time point (onset, peak, recovery) from 3 EAE inductions; 8–10 mice were immunized in each induction.
Figure 7. Affected functions in MiDMs and MDMs at EAE onset. nCounter inflammatory gene expression data were uploaded to IPA. Genes with fold change (EAE onset vs. Naive) ≥1.5 or ≤−1.5 were included in downstream effects analysis. (A) MDMs up-regulated functions, sorted by activation z-score. (B) MiDMs up-regulated (left) and down-regulated (right) functions, sorted by activation z score. The bias term indicates imbalanced numbers of up- and down-regulated genes associated with a distal function requiring significance at the P < 0.01 level. We studied pooled samples from 5 mice in each time point (onset, peak, recovery) from 3 EAE inductions; 8–10 mice were immunized in each induction.CC BY-NC-SA · PubMed Central
Figure 8. Restoration of affected inflammatory genes in resident microglia and recruited monocytes at recovery stage. For each gene, fold change of all different disease stages versus naive state were calculated. Genes that contained at least one fold change >2 or < 0.5 and average fold change for peak and onset >2 or <0.5 were presented. (A) Microglial up-regulated; (B) Microglial down-regulated; (C) monocyte up-regulated; (D) monocyte down-regulated. We studied pooled samples from five mice in each time point (onset, peak, recovery) from three EAE inductions; 8–10 mice were immunized in each induction. FC, fold change.
Figure 8. Restoration of affected inflammatory genes in resident microglia and recruited monocytes at recovery stage. For each gene, fold change of all different disease stages versus naive state were calculated. Genes that contained at least one fold change >2 or < 0.5 and average fold change for peak and onset >2 or <0.5 were presented. (A) Microglial up-regulated; (B) Microglial down-regulated; (C) monocyte up-regulated; (D) monocyte down-regulated. We studied pooled samples from five mice in each time point (onset, peak, recovery) from three EAE inductions; 8–10 mice were immunized in each induction. FC, fold change.CC BY-NC-SA · PubMed Central
Figure 9. Function networks in MiDMs and MDMs. nCounter inflammatory gene expression data were uploaded to IPA. Genes with fold change (EAE onset vs. Naive) ≥1.5 or ≤−1.5 were included in upstream regulators analysis. Predicted upstream regulators were manually curated to form functional clusters. Clusters were uploaded to IPA using Z scores as reference value for each gene. Networks were generated for each cluster consisting of uploaded genes and additional predicted molecules. (A) Typical example of functions with dissimilar activation pattern in MiDMs and MDMs: HIF1A. (B) Function with similar activation pattern in MiDMs and MDMs: Type I IFN. Red object denotes positive (>2) z score and green object denotes negative (<2) z-score. Orange object denotes predicted activation of the network object. Blue object denotes predicted inhibition of the network object. Predicted relationships (connecting lines): orange, leads to activation; blue, leads to inhibition; yellow, finding inconsistent with state of downstream molecule; gray, effect not predicted.
Figure 9. Function networks in MiDMs and MDMs. nCounter inflammatory gene expression data were uploaded to IPA. Genes with fold change (EAE onset vs. Naive) ≥1.5 or ≤−1.5 were included in upstream regulators analysis. Predicted upstream regulators were manually curated to form functional clusters. Clusters were uploaded to IPA using Z scores as reference value for each gene. Networks were generated for each cluster consisting of uploaded genes and additional predicted molecules. (A) Typical example of functions with dissimilar activation pattern in MiDMs and MDMs: HIF1A. (B) Function with similar activation pattern in MiDMs and MDMs: Type I IFN. Red object denotes positive (>2) z score and green object denotes negative (<2) z-score. Orange object denotes predicted activation of the network object. Blue object denotes predicted inhibition of the network object. Predicted relationships (connecting lines): orange, leads to activation; blue, leads to inhibition; yellow, finding inconsistent with state of downstream molecule; gray, effect not predicted.CC BY-NC-SA · PubMed Central

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91 Own result · 24 Own interpretation · 14 Partly supported · 14 Own method · 6 No full text of the sources · 4 Supported · 4 No basis given · 3 Supported so far · 3 Not checked yet · 1 Sources not found · accuracy not yet measured

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  • The paper relies on this without support.

    Monocytes continuously differentiate throughout postnatal life from bone marrow hematopoietic stem cells, which require the transcription factor Myb.

    “whereas monocytes continuously differentiate throughout postnatal life from bone marrow hematopoietic stem cells (HSCs), which require the transcription factor Myb.”

  • The paper relies on this without support.

    Macrophages derived from microglia and monocytes are indistinguishable at the light microscopic level both morphologically and by surface markers.

    “In tissue sections, macrophages derived from these two distinct precursors are indistinguishable at the light microscopic level both morphologically and by surface markers.”

  • The paper relies on this without support.

    There is virtually no background trafficking of monocytes in the CNS parenchyma of healthy animals.

    “as there is virtually no background trafficking of monocytes in the CNS parenchyma of healthy animals.”

  • The authors state this as an assumption.

    The cellular mechanisms by which macrophages promote disease progression are uncertain.

    “However the cellular mechanisms by which macrophages promote disease progression are uncertain.”

  • The paper relies on this without support.

    Microglia represent one instance of tissue-resident macrophages, which reside in all organs.

    “Microglia represent one instance of tissue-resident macrophages, which reside in all organs.”

  • The authors state this as an assumption.

    Exposure of monocytes and microglia to the same intensely inflammatory environment increases the likelihood that they adopt a convergent inflammatory phenotype.

    “to increase the likelihood that ambient conditions could activate these two myeloid cell types toward a convergent inflammatory phenotype”

  • The paper relies on this without support.

    The EAE disease model exposes both recruited monocytes and resident microglia to the same intensely inflammatory environment.

    “Selection of the EAE disease model ensured that both recruited monocytes and resident microglia were exposed to the same intensely inflammatory environment”
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Metadata

Authors
Yamasaki, R., Lu, H., Butovsky, O., Ohno, N., Rietsch, A. M., Cialic, R., Wu, P. M., Doykan, C. E., Lin, J., Cotleur, A. C., Kidd, G., Zorlu, M. M., Sun, N., Hu, W., Liu, L., Lee, J.-C., Taylor, S. E., Uehlein, L., Dixon, D., Gu, J., Floruta, C. M., Zhu, M., Charo, I. F., Weiner, H. L., & Ransohoff, R. M.
Affiliations
  • Cleveland Clinic (US)
  • Brigham and Women's Hospital (US)
  • Vanderbilt University (US)
  • Case Western Reserve University (US)
  • University of California, San Francisco (US)
Corresponding author
Ransohoff, Richard M.
ORCID
Venue
Journal of Experimental Medicine
Publisher
Rockefeller University Press
Published
2014-07-07
Volume
211
Issue
8
Pages
1533-1549
ISSN
  • 0022-1007 (ISSN-L)
  • 1540-9538
DOI
10.1084/jem.20132477
PMC
PMC4113947
OpenAlex
W2159443316
Cited by
856
References
51
Citations by year
2026: 26 · 2025: 49 · 2024: 54 · 2023: 42 · 2022: 58 · 2021: 103 · 2020: 78 · 2019: 76 · 2018: 87 · 2017: 92 · 2016: 96 · 2015: 77 · 2014: 15
FWCI
59.13
Type
journal article
Language
en
Open access
Locations
  • The Journal of Experimental Medicine (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
  • Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
  • eScholarship (California Digital Library) (submittedVersion)
  • Hacettepe University Institutional Repository (hacettepe.edu.tr) (submittedVersion, other-oa)
Funders
  • Japan Society for the Promotion of Science (26860129)
  • National Institute of General Medical Sciences (T32GM007250)
  • National Institute of Neurological Disorders and Stroke (R01NS088137)
Topics
  • Health Sciences › Medicine › Clinical Neurology › Multiple Sclerosis Research Studies
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
Keywords
microglia, monocytes, macrophages, multiple sclerosis, experimental autoimmune encephalomyelitis, demyelination, monocyte-derived macrophages, nodes of Ranvier, phagocytosis, cellular metabolism, tissue damage, neuroinflammation, serial block-face scanning electron microscopy
MeSH terms
CX3C Chemokine Receptor 1, Animals, Central Nervous System, Encephalomyelitis, Autoimmune, Experimental, Gene Expression Regulation, Homeostasis, Humans, Inflammation, Kinetics, Macrophages, Mice, Inbred C57BL, Monocytes, Ranvier's Nodes, Time Factors, Signal Transduction, Microglia, Receptors, Chemokine, Gene Expression Profiling, Cell Shape, Mice, Gene Regulatory Networks, Receptors, CCR2

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.

A language model drew the claims from the paper on 2026-10-16, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).

A second model labelled each claim on 2026-10-16 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).

Cited sources are checked with a language model’s help (gpt-6-sol).

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 ›

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