Journal: Nature immunology

Article Title: Lymphotoxin-dependent elevated meningeal CXCL13:BAFF ratios drive grey matter injury

doi: 10.1038/s41590-025-02359-5

Figure Lengend Snippet: ( a ) qPCR of Lta and Ltb in the leptomeninges of vehicle–treated or BTKi–treated SJL/J adoptive transfer EAE mice at peak disease. ( b-d ) Single-cell RNA sequencing of leptomeninges from young (n=2), and old (n=2) SJL/J mice at acute A/T EAE and appropriate age-matched, naïve controls young and old EAE mice. Data shown represent 2 biological and experimental repeats, each with an n of 1 for each group analyzed. ( b ) Violin plot showing relative gene expression of Btk and lymphotoxin ligand genes ( Lta , Ltb ) in identified cell clusters stratified by age. ( c ) Uniform manifold projection (UMAP) of 16,568 leptomeningeal cells after unsupervised clustering. ( d ) Gene co-expression of Btk with Ltb, Lta projected on the global UMAP. ( e ) Flow cytometry of the LTβ positive population in CD19+ B220+ B cells from naive SJL/J (n=7) or Ltb −/− mice (n=7) splenocytes stimulated with mouse anti-CD40 (5ug/ml) + LPS (1ug/ml) (Stim) ex vivo or pre-treated 1 hour with BTKi (10nM). Data in ( a ) and ( e ) are shown as means ± SD. Statistical analysis was conducted using two-sided unpaired t test for (a) and two-sided Mann-Whitney for (e). ns, not significant. n = 7 in each group.

Article Snippet: InVivo MAb anti-mouse CD40 (BioxCell BE0016–2) (5ug/ml) + LPS (Sigma-Aldrich L2880) (1ug/ml) were added to the culture overnight (18–20 h).

Techniques: Adoptive Transfer Assay, Single Cell, RNA Sequencing, Gene Expression, Expressing, Flow Cytometry, Ex Vivo, MANN-WHITNEY

Journal: Nature immunology

Article Title: Lymphotoxin-dependent elevated meningeal CXCL13:BAFF ratios drive grey matter injury

doi: 10.1038/s41590-025-02359-5

Figure Lengend Snippet: (a) Gating strategy for B cells. (b) Representative plots of LTβ + CD19 + B220 + B cells from naive SJL/J splenocytes either unstimulated or stimulated with mouse anti-CD40 (5ug/ml) + LPS (1ug/ml) ex vivo after pre-treatment with BTKi (10nM) or equivalent Vol of culture medium for 1 hour.

Article Snippet: InVivo MAb anti-mouse CD40 (BioxCell BE0016–2) (5ug/ml) + LPS (Sigma-Aldrich L2880) (1ug/ml) were added to the culture overnight (18–20 h).

Techniques: Ex Vivo

Journal: Mucosal Immunology

Article Title: MicroRNA-142 regulates gut associated lymphoid tissues and group 3 innate lymphoid cells

doi: 10.1016/j.mucimm.2024.09.001

Figure Lengend Snippet: Absence of Mir142 protects against anti-CD40 mediated innate immune-driven colitis. A. Percentage of initial body weight (Day 0 = 100 %) of Mir142 − / − Rag1 − / − mice and Rag1 − / − controls calculated each day after treatment with anti-CD40, or vehicle control (PBS). B. As for (A) but for Daily Disease Activity Index (DAI) scores. C. Representative micrographs of H&E stained distal colon at D8 of treatment of Mir142 − / − Rag1 − / − mice and Rag1 − / − treated with anti-CD40, or vehicle control (PBS). D. Blinded histology scoring of micrographs as in (C). E. Concentrations of TNFα and IFNγ from media of colonic explant culture of Mir142 − / − Rag1 − / − mice and Rag1 − / − treated with anti-CD40 at D8 post-treatment, as determined by ELISA. F. Proportion of ILC3s producing IL-22 following ex vivo PMA/ionomycin re-stimulation, from Mir142 − / − Rag1 − / − mice and Rag1 − / − treated with anti-CD40 at D8 post-treatment. Welch’s t -test, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns = non-significant difference. n = 4–6 mice per experimental group. Data points represent individual biological replicates.

Article Snippet: 150 μg of anti-CD40 mAb (clone FGK4.5, Bio X Cell) or vehicle control (PBS) was administered intraperitoneally to 6- to 8-week-old mice.

Techniques: Control, Activity Assay, Staining, Enzyme-linked Immunosorbent Assay, Ex Vivo

Journal: Cell reports

Article Title: Clostridioides difficile toxin B subverts germinal center and antibody recall responses by stimulating a drug-treatable CXCR4-dependent mechanism

doi: 10.1016/j.celrep.2024.114245

Figure Lengend Snippet: Female B6 mice were given PBS vehicle (left, gray/black, n = 6) or 1 ng TcdB (i.p.) (center and right, n = 10 in total) and then immunized after 5 h with 20 μg of B2Δ/Alum (s.c.). TcdB2-treated mice were injected s.c. with 100 μg isotype control mAb (center, blue n = 5) or 100 μg anti-CD40 mAb (right, green, n = 5) on days 1 and 8. A booster vaccine was administered on day 60 and consisted of 20 μg of B2Δ in PBS. Blood samples were collected before (day 60) and after (day 74) the booster. (A–D) Data show (A) IgM, (B) IgG1, (C) IgG2b, and (D) IgG2c B2Δ-specific endpoint titers ± SD. Matched-pairs t-tests were used to measure significance. (E) Data from (A)–(D) were re-analyzed by calculating fold change (mean ± SD) in endpoint IgM, IgG1, IgG2b, and IgG2c titers following booster vaccine administration and comparing the three experimental groups. Additional ANOVAs with Kruskal-Wallace post-test were performed to compare post-booster titers in all groups. IgG2b overall p = 0.0009, IgG2c overall p = 0.0028. For post-test, ** p < 0.01. (F and G) Representative images of ELISPOT wells with spots attributable to B2Δ-specific IgG1 and IgG2b. Graphs depict the number of B2Δ-specific spots per million cells. Each symbol represents an individual mouse. * p < 0.05, ** p < 0.01.

Article Snippet: InVivoPlus TM anti-mouse CD40 mAb, clone FGK4.5 , BioXCell , Cat# BP0016–2 RRID: AB_1107647.

Techniques: Injection, Control, Enzyme-linked Immunospot

Journal: Cell reports

Article Title: Clostridioides difficile toxin B subverts germinal center and antibody recall responses by stimulating a drug-treatable CXCR4-dependent mechanism

doi: 10.1016/j.celrep.2024.114245

Figure Lengend Snippet:

Article Snippet: InVivoPlus TM anti-mouse CD40 mAb, clone FGK4.5 , BioXCell , Cat# BP0016–2 RRID: AB_1107647.

Techniques: Control, Virus, Recombinant, Expressing, Suspension, Protease Inhibitor, SYBR Green Assay, Cell Isolation, Binding Assay, Bradford Protein Assay, Cell Counting, Enzyme-linked Immunosorbent Assay, Gene Expression, Software, Simple Western, Protein Extraction