Journal: Molecular Neurodegeneration

Article Title: NOTCH2NLC GGC intermediate repeat with serine induces hypermyelination and early Parkinson’s disease-like phenotypes in mice

doi: 10.1186/s13024-024-00780-2

Figure Lengend Snippet: Mitochondrial-related pathways and myelin sheath component genes are dysregulated in N2C-32G13S mice. Cortical tissues from 12-month-old N2C transgenic mice were used for bulk RNA sequencing. n = 3 mice per group. ( A ) Volcano plot showing differentially expressed genes between N2C-32G13S and N2C-45G mice at padj < 0.05 and fold change < 0.67 or > 1.5. Top dysregulated genes are highlighted, with those related to mitochondrial function shown in blue. ( B ) For differentially expressed genes between N2C-32G13S and N2C-45G mice, 54 top dysregulated genes are shown in biological process (BP), cellular component (CC), and molecular function (MF) by gene ontology (GO) term analysis. ( C ) Top dysregulated KEGG pathways between N2C-32G13S and N2C-45G mice. ( D ) Connections between top differentially expressed genes and cellular components. Genes highlighted in red are myelin sheath components. ( E-H ) Quantitative PCR was performed for myelin sheath component genes MBP ( E ), MOG ( F ), GJC2 ( G ), and ERMN ( H ) using cortical tissues from 12-month-old N2C mice. ( I ) Cortical tissues were used for immunoblotting. ( J-K ) Quantification of MBP ( J ) and MOG ( K ) from western blot in I . Data are presented as the mean ± SD, n = 3. * P < 0.05 and ** P < 0.01 by one-way ANOVA with Tukey’s post hoc test

Article Snippet: The antibodies used in this study were tyrosine hydroxylase (TH) (Millipore, #MAB318), LRRK2 (Abcam, #ab133474), α-synuclein (BD Transduction Lab, #610787), p62 (Cell Signaling, #39749), DRP1 (Cell Signaling, #8570), LC3II (Abcam, #ab243506), CD11b (Cell Signaling, #49420S), MBP (Cell Signaling, #83683), MOG (Merck Millipore, MAB5680), β-actin (Santa Cruz, #AC-15).

Techniques: Transgenic Assay, RNA Sequencing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Frontiers in Immunology

Article Title: Dual role of peripheral B cells in multiple sclerosis: emerging remote players in demyelination and novel diagnostic biomarkers

doi: 10.3389/fimmu.2023.1224217

Figure Lengend Snippet: Myelin markers decrease induced by B cell-derived EVs containing myelin antibodies in oligodendrocytes in vitro . (A) Experimental protocol of in vitro study. Oligodendrocytes were plated and cultured using a seeding density of 1 × 10 4 cells/cm 2 and grown to 70% confluence. At 7 days, the cells received 100 μg of B-cell-derived EVs for 7 days. After fixing, myelin markers were analyzed. (B) Representative immunofluorescence images of oligodendrocytes expressing MBP, MOG, and CNP after receiving 100 µg of B-cell-derived EVs from healthy controls, white matter lesion controls, and MS cases in an in vitro assay. 4′,6-diamidino-2-phenylindole (DAPI) was used for nuclear staining. Quantitative analysis of MBP, MOG, and CNP marker expression by immunofluorescence ( n = 10 assays per group). Data are mean ± SD. * p < 0.05, **p < 0.01, ***p < 0.001. (C) Quantitative analysis of cell viability after EV administration. Data are mean percentage ± SD. AU, arbitrary unit; CNP, 2’,3’-cyclic-nucleotide 3’-phosphodiesterase; DAPI, 4 ‘,6-diamidino-2-fenilindol; HCs, healthy controls; EVs, extracellular vesicles; MBP, myelin basic protein; MOG, myelin oligodendrocyte glycoprotein; MS, multiple sclerosis; WMC, white matter lesion controls.

Article Snippet: Anti-myelin basic protein (MBP), anti-MOG antibodies and anti-albumin (as negative control) were detected and quantified by sandwich enzyme-linked immunosorbent assay (ELISA) (Abbexa, UK).

Techniques: Derivative Assay, In Vitro, Cell Culture, Immunofluorescence, Expressing, Staining, Marker

Journal: Frontiers in Immunology

Article Title: Dual role of peripheral B cells in multiple sclerosis: emerging remote players in demyelination and novel diagnostic biomarkers

doi: 10.3389/fimmu.2023.1224217

Figure Lengend Snippet: Myelin antibody content in blood and cerebrospinal fluid. (A) Graph showing soluble anti-MBP and anti-MOG antibodies in healthy controls (n = 39), white matter lesion controls (n = 23), and MS cases (n = 136). (B) Comparison of anti-MBP and anti-MOG antibody content in total circulating EVs between healthy controls, white matter lesion controls, and MS cases. (C) Chart comparing myelin antibody content in total circulating EVs and B-cell-derived EVs. (D) Myelin antibody content in B-cell-derived EVs from CSF and blood. Data are mean ± SD. * p <0.05. ABs, antibodies; CSF, cerebrospinal fluid; EVs, extracellular vesicles; MBP, myelin basic protein; MOG, myelin oligodendrocyte glycoprotein; MS, multiple sclerosis.

Article Snippet: Anti-myelin basic protein (MBP), anti-MOG antibodies and anti-albumin (as negative control) were detected and quantified by sandwich enzyme-linked immunosorbent assay (ELISA) (Abbexa, UK).

Techniques: Comparison, Derivative Assay

Journal: Frontiers in Immunology

Article Title: Dual role of peripheral B cells in multiple sclerosis: emerging remote players in demyelination and novel diagnostic biomarkers

doi: 10.3389/fimmu.2023.1224217

Figure Lengend Snippet: Myelin antibody content in B cell-derived EVs. (A) B cell-derived EV myelin antibody content in healthy controls (n = 39) and patients with MS (n = 136). (B) Receiver operating characteristic curves representing the cut-off point corresponding to the maximum sensitivity and specificity values in order to identify the myelin antibody levels to differentiate MS cases and healthy controls. (C) Correlation between EV anti-MBP and anti-MOG antibody content in MS patients. (D) Receiver operating characteristic curves representing the combination of anti-MBP and anti-MOG antibodies from B cell-derived EVs representing a cut-off point corresponding to the maximum sensitivity and specificity values to differentiate the MS patients from healthy controls. (E) Analysis of EV anti-albumin antibody content in MS patients and in healthy control. (F) Oligodendrocyte-derived EV myelin antibody content in healthy controls (n = 39) and patients with MS (n = 136). (G) Receiver operating characteristic curves representing the cut-off point corresponding to the maximum sensitivity and specificity values in order to identify the MBP antibody levels in oligodendrocyte-derived extracellular vesicles to differentiate MS cases and healthy controls.

Article Snippet: Anti-myelin basic protein (MBP), anti-MOG antibodies and anti-albumin (as negative control) were detected and quantified by sandwich enzyme-linked immunosorbent assay (ELISA) (Abbexa, UK).

Techniques: Derivative Assay, Control

Journal: Frontiers in Immunology

Article Title: Dual role of peripheral B cells in multiple sclerosis: emerging remote players in demyelination and novel diagnostic biomarkers

doi: 10.3389/fimmu.2023.1224217

Figure Lengend Snippet: Impact of disease modifying treatments and disease activity in EVs anti-myelin antibodies content. (A) Myelin antibody content from B cell-derived EVs in treated patients with MS (n = 54), naïve patients (n = 82) and healthy controls (n=39). (B) Myelin antibody content from B cell-derived EVs in treated MS patients with anti-CD20 therapies compared with treated MS patients with different therapies. (C) Anti-MBP and anti-MOG levels of B cell-derived EVs, comparing patients with MS with active (n = 77) and stable disease (n = 59). Data are mean ± SD. ***p < 0.001. AUC, area under curve; EVs, extracellular vesicles; MBP, myelin basic protein; MOG, myelin oligodendrocyte glycoprotein; MS, multiple sclerosis.

Article Snippet: Anti-myelin basic protein (MBP), anti-MOG antibodies and anti-albumin (as negative control) were detected and quantified by sandwich enzyme-linked immunosorbent assay (ELISA) (Abbexa, UK).

Techniques: Activity Assay, Derivative Assay