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 is highly expressed on infiltrating myeloid cells in the CNS throughout RR-EAE. RR-EAE was induced in female SJL/J mice using PLP 139–151 emulsified in complete Freund’s adjuvant. Spleen and CNS tissues were collected at defined disease stages and analyzed for Ninjurin-1 expression by flow cytometry. (A) RR-EAE clinical disease course (n = 8). (B) CD45 hi B220 - CD3 - CD11b + infiltrating myeloid cells in the CNS displayed a marked increase in Ninjurin-1 expression compared with splenic myeloid cells at all disease stages (pre-onset, onset, peak, remission, and relapse). (C) CD45 hi CD3 + CD11b - T cells in the CNS displayed consistently low Ninjurin-1 expression, with no significant differences compared to splenic CD3 + T cells at any disease stage. All time points include n = 4 mice. Data are representative of two independent EAE experiments. Results are shown as mean ± SEM and were analyzed using two-way ANOVA (*p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: Mice received three subcutaneous flank injections (total 100 μL) of PLP 139–151 peptide (200 μg/mouse; Genemed Biotechnologies Inc.) emulsified in Complete Freund’s Adjuvant (BD Biosciences) containing 200 μg of Mycobacterium tuberculosis H37Ra.

Techniques: Adjuvant, Expressing, Flow Cytometry

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: Anti-Ninj 26–37 treatment at peak RR-EAE attenuates relapse and reduces CNS immune cell infiltration. RR-EAE was induced in SJL/J mice by subcutaneous injection of PLP 139–151 emulsified in CFA. Mice were treated intraperitoneally three times per week with anti-Ninj 26–37 peptide (1 mg/mL; n = 11) or scramble control (n = 11), starting at the peak of disease (day 15) and continuing until day 30 post-induction. (A) Anti-Ninj 26–37 –treated mice (open circles) displayed significantly reduced clinical scores and were protected from relapse compared with scramble-treated controls (closed squares; ***p < 0.001 by Mann–Whitney test). (B) Flow cytometry of CNS mononuclear cells at day 32 post-immunization revealed a marked reduction in total CD45 hi leukocytes, including CD3 + T cells, CD4 + T cells, B220 + B cells, and CD11b + infiltrating myeloid cells, in anti-Ninj 26-37 -treated mice. Absolute cell numbers were calculated by multiplying the frequency of each lineage-positive population determined by flow cytometry by the total number of mononuclear cells recovered from the CNS of each mouse. All CNS samples were processed using identical digestion and purification procedures prior to flow cytometric analysis (n = 5; *p < 0.05 by multiple unpaired t-tests). (C) Representative H&E-stained spinal cord sections collected at day 30 show reduced inflammatory infiltrates in anti-Ninj 26–37 –treated mice compared to controls. Data are representative of at least three sections per mouse from four mice per group. Scale bar = 200 µm. (D) Cellular infiltration was quantified by counting hematoxylin-positive nuclei within standardized 300 × 300 µm white matter regions (three ROIs per section, averaged per mouse) and expressed as cells per mm². Each dot represents one mouse; bars indicate mean ± SEM (n = 3; **p < 0.01 by unpaired t-tests).

Article Snippet: Mice received three subcutaneous flank injections (total 100 μL) of PLP 139–151 peptide (200 μg/mouse; Genemed Biotechnologies Inc.) emulsified in Complete Freund’s Adjuvant (BD Biosciences) containing 200 μg of Mycobacterium tuberculosis H37Ra.

Techniques: Injection, Control, MANN-WHITNEY, Flow Cytometry, Purification, Staining