Journal: Frontiers in Immunology

Article Title: Dual role of Ninjurin-1 in myeloid cell adhesion and inflammation in relapse-remitting EAE

doi: 10.3389/fimmu.2026.1803382

Figure Lengend Snippet: Ninjurin-1 + myeloid cells enhance CD4 + T cell activation and cytokine production. CD45 + B220 - CD3 - CD11b + Ly6G - Ninjurin-1+ or Ninjurin-1 - myeloid cells were flow-sorted and co-cultured with CFSE-labeled CD4 + T cells isolated from 2D2 mice. Cells were stimulated with MOG 35–55 peptide in 96-well plates for 72 hours. (A) T cell proliferation, measured by CFSE dilution, was greater when CD4 + T cells were co-cultured with Ninjurin-1 + myeloid cells compared to Ninjurin-1 - cells. Controls included CD4 + T cells cultured without antigen-presenting cells (APCs) or without MOG 35–55 peptide. (B) CD4 + T cells co-cultured with Ninjurin-1 + myeloid cells showed increased expression of CD25 and TNF-α, indicating higher levels of activation and inflammatory cytokine production. Data represent n = 4 independent experiments and are shown as mean ± SEM. Statistical analysis was performed using two-way ANOVA (*p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: CFSE + CD4 + T cells (100,000/well) were then co-cultured with either Ninjurin-1 - or Ninjurin-1 + sorted cells (40,000/well) with 20 μg/ml MOG 35–55 peptide (Genemed Biotechnologies Inc.) into a 96 well plate for 72 hours.

Techniques: Activation Assay, Cell Culture, Labeling, Isolation, Expressing

Journal: bioRxiv

Article Title: The Glucose Transporter GLUT3 Controls Regulatory T Cell Function

doi: 10.64898/2026.03.26.714439

Figure Lengend Snippet: (A and B) Relative body weight changes (A) and experimental autoimmune encephalomyelitis (EAE) clinical scores (B) in WT, GLUT1-deficient ( Slc2a1 fl/fl Cd4 Cre ) and GLUT3-deficient ( Slc2a3 fl/fl Cd4 Cre ) mice following immunization with MOG 35-55 peptide emulsified in CFA; means ± SEM of 6-11 mice per cohort. (C) RT-qPCR analysis of Slc2a1 (GLUT1) and Slc2a3 (GLUT3) expression in isolated splenic CD4 + T cells from WT, GLUT1-deficient and GLUT3-deficient mice used in EAE experiments; data are shown as means ± SEM of 4-11 mice. (D and E) Representative flow cytometric analysis of Foxp3 + Treg cells in the central nervous system (CNS) of WT, GLUT1- and GLUT3-deficient mice 20 days after MOG 35-55 immunization (D) , with quantification of Treg cell frequencies and absolute cell numbers (E) ; means ± SEM of 6-11 mice. (F and G) Frequencies of GM-CSF and IL-2-producing CD4 + T cells in the CNS of WT, GLUT1-and GLUT3-deficient mice 20 days after immunization with MOG 35-55 peptide and restimulation with PMA/ionomycin for 5 h (F) , with quantification of total cell numbers of GM-CSF and IL-2 producing CD4 + T cells in the CNS (G) ; means ± SEM of 6-11 mice. Statistical analyses shown in (A, B, C, E, G) were performed using two-way ANOVA. *, p<0.05; **, p<0.01; ***, p<0.001; ns, non-significant.

Article Snippet: EAE was induced by emulsifying 2 mg/mL myelin MOG 35–55 peptide (Synpeptide) in incomplete Freund’s adjuvant (IFA; Fisher Scientific) supplemented with 5 mg/mL Mycobacterium tuberculosis H37Ra (Fisher Scientific) via syringe extrusion.

Techniques: Quantitative RT-PCR, Expressing, Isolation

Journal: International Journal of Molecular Sciences

Article Title: Early Reduction in Mitochondrial Membrane Potential in Synaptic Mitochondria Contribute to Synaptic Pathology in the EAE Mouse Model of Multiple Sclerosis

doi: 10.3390/ijms27062579

Figure Lengend Snippet: Decreased expression of MIC60 in synaptic mitochondria of photoreceptor ribbon synapses in the OPL of the retina in MOG/CFA-injected EAE mice in comparison to CFA-injected control mice at day 5 after injection. 0.5 µm thin retinal sections from CFA control mice ( A1 – A3 , C1 – C3 ) and EAE mice ( B1 – B3 , D1 – D3 ) double-immunolabelled with rabbit polyclonal anti-MIC60 antibody and mouse monoclonal anti-actin antibody. Merged images from green ( A1 , B1 , C1 , D1 ) and red channels ( A2 , B2 , C2 , D2 ) were overlayed in ( A3 , B3 , C3 , D3 ). ( A1 – A3 , B1 – B3 ) show lower magnified representative images. Higher magnification images from the immunolabelled OPL are shown in ( C1 – C3 , D1 – D3 ). An exemplary region-of-interest (ROI) covering the OPL, in which the photoreceptor synapses are located, is schematically plotted in orange with a dashed line in ( A1 – A3 , B1 – B3 ). ( E , F ) Quantification of MIC60 and actin immunosignals in the OPL (measured as integrated density; data depicted as SuperPlots). MOG/CFA values were normalised to the corresponding actin signal in the OPL and related to the arithmetic mean of CFA, which was set to 100% in each independent experiment ( E , F ). The Superplots show the arithmetic means (green horizontal lines) ± S.E.M.s. Filled colored circles represent the arithmetic means of the individual independent experiments; the open circles show the individual datapoints from all experiments. One-sample t -tests were used for statistical analyses in ( E , F ). p -values < 0.05 were considered statistically significant. Abbreviations: CFA, complete Freund’s adjuvant; MOG, myelin oligodendrocyte glycoprotein; EAE, experimental autoimmune encephalomyelitis; IS, inner segment; OPL, outer plexiform layer; ONL, outer nuclear layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer; S.E.M., standard error of the mean; N = number of independent experiments; n = number of analysed images; n.s., non-significant; *, p < 0.05. Scale bars: 5 μm.

Article Snippet: On the same day, ~1–2 h after MOG/CFA or CFA only injection, and on the next day, both EAE and control mice were injected intraperitoneally with pertussis toxin (PTX) (List Biological Laboratories via Biozol (Eching, Germany #181)).

Techniques: Expressing, Injection, Comparison, Control, Adjuvant

Journal: International Journal of Molecular Sciences

Article Title: Early Reduction in Mitochondrial Membrane Potential in Synaptic Mitochondria Contribute to Synaptic Pathology in the EAE Mouse Model of Multiple Sclerosis

doi: 10.3390/ijms27062579

Figure Lengend Snippet: Decreased expression of MIC60 and RIBEYE in photoreceptor ribbon synapses in the OPL of the retina in MOG/CFA-injected EAE mice in comparison to CFA-injected control mice at day 5 after injection. The 0.5 µm thin retinal sections from CFA control mice ( A1 – A3 , C1 – C3 ) and EAE mice ( B1 – B3 , D1 – D3 ) were double-immunolabelled with rabbit polyclonal anti-MIC60 antibody and mouse monoclonal anti-RIBEYE antibody (clone 2D9). Merged images from green ( A1 , B1 , C1 , D1 ) and red channels ( A2 , B2 , C2 , D2 ) were overlayed in ( A3 , B3 , C3 , D3 ). ( A1 – A3 , B1 – B3 ) shows lower magnified representative images. Higher magnification images from the immunolabelled OPL are shown in ( C1–C3,D1–D3 ). An exemplary region-of-interest (ROI) covering the OPL, in which the photoreceptor synapses are located, is schematically plotted in orange with a dashed line in ( A1 – A3 , B1 – B3 ). ( E , F ) Quantification of MIC60 and RIBEYE immunosignals in the OPL (measured as integrated density; data depicted as SuperPlots). MOG/CFA values were normalised to the arithmetic means of CFA, which were set to 100% in each independent experiment ( E , F ). The Superplots show the arithmetic means (green horizontal lines) ± S.E.M.s. Filled circles represent the arithmetic means of the individual independent experiments; the open circles show the individual datapoints from all experiments. One-sample t -tests were used for statistical analyses in ( E , F ). Abbreviations: CFA, complete Freund’s adjuvant; MOG, myelin oligodendrocyte glycoprotein; EAE, experimental autoimmune encephalomyelitis; OS, outer segment; IS, inner segment; OPL, outer plexiform layer; ONL, outer nuclear layer; S.E.M., standard error of the mean; N = number of independent experiments; n = number of analysed images; ***, p < 0.001 **, p < 0.01. Scale bars: 5 μm.

Article Snippet: On the same day, ~1–2 h after MOG/CFA or CFA only injection, and on the next day, both EAE and control mice were injected intraperitoneally with pertussis toxin (PTX) (List Biological Laboratories via Biozol (Eching, Germany #181)).

Techniques: Expressing, Injection, Comparison, Control, Adjuvant

Journal: International Journal of Molecular Sciences

Article Title: Early Reduction in Mitochondrial Membrane Potential in Synaptic Mitochondria Contribute to Synaptic Pathology in the EAE Mouse Model of Multiple Sclerosis

doi: 10.3390/ijms27062579

Figure Lengend Snippet: Differences in optokinetic responses on day 5 after injection in MOG/CFA-injected EAE mice in comparison to CFA-injected control mice. Optokinetic responses were recorded from MOG/CFA-injected EAE mice on day 5 after injection and analysed for frequency thresholds ( A ) and contrast sensitivity/threshold at two different rotation speeds: 0.192 cycles/degree ( B1 , B2 ) and 0.272 cycles/degree ( C1 , C2 ). Data are depicted as SuperPlots. The SuperPlots show the arithmetic means (green horizontal lines) ± S.E.M.s. Filled circles represent the arithmetic means of the individual independent experiments; the open circles show the individual datapoints from all experiments. Welch’s t -test ( t -test with Welch’s correction) was used for statistical analyses in ( A , B1 , B2 ); Mann–Whitney test for ( C1 , C2 ). p -values < 0.05 were considered statistically significant. Abbreviations: EAE, experimental autoimmune encephalomyelitis; S.E.M., standard error of the mean; N, number of independent mice; n = number of independent measurements; ****, p < 0.0001; ***, p < 0.001; n.s., non-significant.

Article Snippet: On the same day, ~1–2 h after MOG/CFA or CFA only injection, and on the next day, both EAE and control mice were injected intraperitoneally with pertussis toxin (PTX) (List Biological Laboratories via Biozol (Eching, Germany #181)).

Techniques: Injection, Comparison, Control, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Early Reduction in Mitochondrial Membrane Potential in Synaptic Mitochondria Contribute to Synaptic Pathology in the EAE Mouse Model of Multiple Sclerosis

doi: 10.3390/ijms27062579

Figure Lengend Snippet: MIC60 IF immunosignals in the OPL are not altered in MOG/CFA-injected mice in comparison to CFA-injected mice at day 3 after injection. The 0.5 µm thin retinal sections from CFA control mice ( A1 – A3 , C1 – C3 ) and EAE mice ( B1 – B3 , D1 – D3 ) obtained at day 3 after injection were double-immunolabelled with rabbit polyclonal anti-MIC60 antibody and mouse monoclonal anti-actin antibody (clone C4 ). Merged images from green ( A1 , B1 , C1 , D1 ) and red channels ( A2 , B2 , C2 , D2 ) were overlayed in ( A3 , B3 , C3 , D3 ). ( A1 – A3 , B1 – B3 ) shows representative lower magnified images. Higher magnification images from the immunolabelled OPL are shown in ( C1–C3,D1 – D3 ). ( E ) The MIC60 IF signal strength in the OPL (measured as integrated density) was normalised to the corresponding actin signal in the OPL and related to the arithmetic means of the CFA values, which were set to 100% in each experiment. ( F ) The actin IF signal measured in OPL (as integrated density) served as a reference protein. Data in ( E , F ) were depicted as SuperPlots. The SuperPlots show the arithmetic means (green horizontal lines) ± S.E.M.s. Filled circles represent the arithmetic means of the individual independent experiments; the open circles show the individual datapoints from all experiments. Wilcoxon signed-rank tests were used for statistical analysis in ( E , F ). p -values < 0.05 were considered statistically significant. Abbreviation: CFA, complete Freund’s adjuvant; MOG, myelin oligodendrocyte glycoprotein; EAE, experimental autoimmune encephalomyelitis; IF, immunofluorescence; IS, inner segment; OPL, outer plexiform layer; ONL, outer nuclear layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer; S.E.M., standard error of the mean; N = number of independent experiments; n = number of analysed images; n.s., non-significant. Scale bars: 5 μm.

Article Snippet: On the same day, ~1–2 h after MOG/CFA or CFA only injection, and on the next day, both EAE and control mice were injected intraperitoneally with pertussis toxin (PTX) (List Biological Laboratories via Biozol (Eching, Germany #181)).

Techniques: Injection, Comparison, Control, Adjuvant, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Early Reduction in Mitochondrial Membrane Potential in Synaptic Mitochondria Contribute to Synaptic Pathology in the EAE Mouse Model of Multiple Sclerosis

doi: 10.3390/ijms27062579

Figure Lengend Snippet: No differences in optokinetic responses on day 3 after injection in MOG/CFA-injected EAE mice in comparison to CFA-injected control mice on day 3 after injection. Optokinetic responses were recorded from MOG/CFA-injected mice on day 3 after injection and analysed for contrast sensitivity/threshold at two different rotation speeds: 0.192 cycles/degree ( A1 , A2 ) and 0.272 cycles/degree ( B1 , B2 ). ( A1 , A2 , B1 , B2 ) Data are depicted as SuperPlots. The SuperPlots show the arithmetic means (green horizontal lines) ± S.E.M.s. Filled circles represent the arithmetic means of the individual independent experiments; the open circles show the individual datapoints from all experiments. Welch’s t -test ( t -test with Welch’s correction) was used for statistical analyses in ( A1 , A2 , B1 , B2 ). p -values < 0.05 were considered statistically significant. Abbreviations: S.E.M., standard error of the mean; N, number of independent mice; n = number of independent measurements; n.s., non-significant.

Article Snippet: On the same day, ~1–2 h after MOG/CFA or CFA only injection, and on the next day, both EAE and control mice were injected intraperitoneally with pertussis toxin (PTX) (List Biological Laboratories via Biozol (Eching, Germany #181)).

Techniques: Injection, Comparison, Control

Journal: International Journal of Molecular Sciences

Article Title: Early Reduction in Mitochondrial Membrane Potential in Synaptic Mitochondria Contribute to Synaptic Pathology in the EAE Mouse Model of Multiple Sclerosis

doi: 10.3390/ijms27062579

Figure Lengend Snippet: Decreased TMRM fluorescence signals in the OPL of MOG/CFA-injected EAE mice in comparison to CFA-injected control mice at day 5 after injection under resting conditions. Representative exemplary images of living retinal slices from CFA control mice ( A1 – A3 , C1 – C3 ) and MOG/CFA-injected mice ( B1 – B3 , D1 – D3 ) that were labelled with TMRM at resting conditions (incubation in AMES medium). ( A1 , B1 , C1 , D1 ) show the TMRM signals; ( A2 , B2 , C2 , D2 ) the phase contrast images. The signals from ( A1 – A3 , C1 – C3 ) and ( B1 – B3 , D1 – D3 ) were overlaid in ( A3 , B3 , C3 , D3 ). In ( C1 – C3 , D1 – D3 ), the respective slices were incubated with the mitochondrial uncoupling agent CCCP to verify the mitochondrial membrane potential as the specific source of the TMRM signals. ( E ) Quantification of the TMRM signals in the OPL of MOG/CFA-injected animals in comparison to CFA-injected control mice. MOG/CFA values were normalised to the arithmetic means of the CFA values, which were set to 100% in each independent experiment. Data are depicted as SuperPlots. The SuperPlots show the arithmetic means (green horizontal lines) ± S.E.M.s. Filled colored circles represent the arithmetic means of the individual independent experiments; the open circles show the individual datapoints from all experiments. Statistical analysis in ( E ) was performed with Wilcoxon signed-rank test. p < 0.05 was considered statistically significant. Abbreviations: OS, outer segment; IS, inner segment; OPL, outer plexiform layer; ONL, outer nuclear layer; N = number of independent experiments; n = number of analysed images; *, p < 0.05. Scale bars: 50 μm.

Article Snippet: On the same day, ~1–2 h after MOG/CFA or CFA only injection, and on the next day, both EAE and control mice were injected intraperitoneally with pertussis toxin (PTX) (List Biological Laboratories via Biozol (Eching, Germany #181)).

Techniques: Fluorescence, Injection, Comparison, Control, Incubation, Membrane

Journal: International Journal of Molecular Sciences

Article Title: Early Reduction in Mitochondrial Membrane Potential in Synaptic Mitochondria Contribute to Synaptic Pathology in the EAE Mouse Model of Multiple Sclerosis

doi: 10.3390/ijms27062579

Figure Lengend Snippet: (A–G) Representative images of TMRM fluorescence signals (mitochondrial membrane potential) and SypHy (exocytosis) signals in retinal slices from MOG/CFA-injected (EAE) SypHy mice and CFA-injected (control) SypHy mice obtained on day 5 after injection. Signals were recorded at different synaptic activity levels of the photoreceptor synapse in the OPL, i.e., during resting conditions, depolarisation and after treatment with CCCP. ( A ) Schematic summary of the recording experiment. ( B1 – B4 , C1 – C4 , D1 – D4 , E1 – E4 , F1 – F4 , G1 – G4 ) Representative images of the corresponding living retinal slices from CFA-injected control mice ( B1 – B3 , C1 – C3 , D1 – D3 ) and MOG/CFA-injected EAE mice ( E1 – E3 , F1 – F3 , G1 – G3 ) at the indicated incubation conditions. ( B1 , C1 , D1 , E1 , F1 , G1 ) show the SypHy signals in the green channel; ( B2 , C2 , D2 , E2 , F2 , G2 ) show the TMRM signals in the red channel; the phase contrast images are shown in ( A3 , B3 , C3 , D3 , E3 , F3 , G3 ). The signals from ( B1 – B3 , C1 – C3 , D1 – D3 , E1 – E3 , F1 – F3 , G1 – G3 ) were overlaid in ( B4 , C4 , D4 , E4 , F4 , G4 ). In ( D1 – D4 , G1 – G4 ), the respective slices were incubated with the mitochondrial uncoupling agent CCCP to verify the mitochondrial membrane potential as a specific source of the TMRM signals. The colored arrows in ( A ) indicate the time points at which the respective signals were used for quantification see ( H – K ). Abbreviations: OPL, outer plexiform layer; RS, resting solution; DS, depolarisation solution. Scale bars: 50 μm. ( H – K ) Decreased TMRM fluorescence signals in the OPL of MOG/CFA-injected EAE mice in response to depolarisation on day 5 after injection, indicating enhanced dissipation of mitochondrial membrane potential in synaptic mitochondria of EAE mice. ( H ) The line graphs show the SypHy signals of the indicated mice recorded from the OPL during incubation in resting solution (in the first 60 s of incubation) and during depolarisation induced by adding a high K + -containing depolarisation solution (DS) (in the following 60 s). ( I1 ) Same incubations as ( H ), but showing the TMRM signals recorded during resting conditions and during depolarisation. In ( I2 ), TMRM values during depolarisation were normalised to the starting point of the depolarisation (F0, set to 1.00). ( J , K ) Quantification of the TMRM signals in the OPL of MOG-/CFA-injected animals in comparison to CFA-injected control mice under depolarisation. Data in ( J , K ) are depicted as SuperPlots. The SuperPlots show the arithmetic means (green horizontal lines) ± S.E.M.s. Filled circles represent the arithmetic means of the individual independent experiments; the open circles show the individual datapoints of all experiments. ( J ) shows normalised TMRM signals at the end of depolarisation. MOG/CFA values are normalised to the arithmetic mean of all CFA experiments (the latter set to 100%). ( K ) displays the depolarisation-induced decrease in TMRM signals normalised to F0. Statistical analyses in ( J , K ) were performed with Welch’s t -test ( t -test with Welch’s correction). p < 0.05 was considered statistically significant. Abbreviation: CFA, complete Freund’s adjuvant; CCCP, carbonyl cyanid-m-chlorphenyl hydrazon; DS, depolarisation solution; MOG, myelin oligodendrocyte glycoprotein; EAE, experimental autoimmune encephalomyelitis; OPL, outer plexiform layer; S.E.M., standard error of the mean; TMRM, Tetramethylrhodamine, methylester; N = number of independent experiments; n = number of analysed images; *, p < 0.05; ***; p < 0.001.

Article Snippet: On the same day, ~1–2 h after MOG/CFA or CFA only injection, and on the next day, both EAE and control mice were injected intraperitoneally with pertussis toxin (PTX) (List Biological Laboratories via Biozol (Eching, Germany #181)).

Techniques: Fluorescence, Membrane, Injection, Control, Activity Assay, Incubation, Comparison, Adjuvant

Journal: Nature

Article Title: Facile induction of immune tolerance by an interleukin-2–TGFβ surrogate agonist

doi: 10.1038/s41586-026-10208-0

Figure Lengend Snippet: a , Schematic illustration of MOG 35–55 and protein administration. Created in BioRender; Sun, Q. https://BioRender.com/j3crwdr (2026). b , c , Representative flow cytometry plots ( b ) and quantification of FOXP3 + and RORγt + FOXP3 + cell frequencies ( b ) and absolute numbers ( c ) among donor CD44 + Ki-67 + 2D2 cells in the indicated lymphoid organs ( n = 8 samples per group). d – h , Therapeutic effect of TGM1–IL-2 in establishing tolerance to suppress MOG 35–55 -induced EAE ( n = 11 mice per group). d , Schematic illustration. CNS, central nervous system; PTX, pertussis toxin; SC, spinal cord; CFA, complete Freund’s adjuvant. Created in BioRender; Sun, Q. https://BioRender.com/t3trkm2 (2026). e , Quantification of mean EAE scores. f , Quantification of CD4 + T cell absolute numbers in the spinal cord. g , Quantification of IFNγ- and IL-17A-producing CD4 + T cell numbers in the spinal cord. h , Representative flow cytometry plots and quantification of GM-CSF + CD4 + T cell frequencies and absolute numbers in the spinal cord. Data are presented as mean ± s.e.m. Data in b , c are pooled from two independent experiments. Data in d – h are representative of two independent experiments. Statistics were obtained by one-way ANOVA coupled with Dunnett’s multiple-comparisons test ( b , c ), two-way ANOVA coupled with Šídák’s multiple-comparisons test ( e ) or unpaired Welch’s t -test (two-tailed) ( f – h ).

Article Snippet: Beginning one day after transfer, mice received 40 μg MOG 35–55 peptide (Genemed Synthesis) together with 50 pmol of the indicated proteins by intraperitoneal injection every other day, for a total of six doses.

Techniques: Flow Cytometry, Adjuvant, Two Tailed Test

Journal: Nature

Article Title: Facile induction of immune tolerance by an interleukin-2–TGFβ surrogate agonist

doi: 10.1038/s41586-026-10208-0

Figure Lengend Snippet: a , Gating strategy for identifying MOG 35-55 -reactive 2D2 cells. b , Representative flow cytometry plots and quantification of expression of the indicated markers in splenic 2D2 cells (n = 6 samples/group). c–e , Therapeutic effect of TGM1–IL-2 in establishing tolerance to suppress MOG 35–55 -induced EAE (n = 11 mice/group). c , Individual EAE scores for mice in the indicated groups. d , Quantification of the numbers of the indicated cell populations in the spinal cord. e , Representative flow cytometry plots and quantification of IFN-γ + and IL-17a + CD4 + T-cell frequencies in the spinal cord. Data are presented as mean ± s.e.m. The data in b–e are representative of two independent experiments. The statistics were obtained by unpaired Welch’s t-test (two-tailed) ( b,d and e ).

Article Snippet: Beginning one day after transfer, mice received 40 μg MOG 35–55 peptide (Genemed Synthesis) together with 50 pmol of the indicated proteins by intraperitoneal injection every other day, for a total of six doses.

Techniques: Flow Cytometry, Expressing, Two Tailed Test