(A) After EAE induction of WT mice, 500 ng/mice rhCTRP4 or control (BSA) were administered intraperitoneally every day starting at disease onset on day 9 after immunization until sacrificed. The clinical scores were monitored daily and are depicted (n = 10/group). (B) Representative images of H&E and LFB staining on spinal cord sections of mice at the peak of EAE. Scale bars: 200 μm. (C) CNS monocytes were harvested on day 18. The absolute cell numbers of indicated CNS-infiltrating cell populations (gated on CD45⁺) or the absolute numbers of CD4⁺IFN-γ⁺, CD4⁺IL-17A⁺, and CD4⁺IL-17A⁺IFN-γ⁺ in CNS were analyzed. (D) Representative immunofluorescent images and quantification of IL-17A⁺ cells in the spinal cord of indicated mice at day 18 after immunization. Nuclei were counterstained with DAPI. Scale bars: 100 μm. (E) Representative immunohistochemistry detection and quantification of p-STAT3(Y705) in the spinal cords of indicated mice at day 18 after immunization. Scale bars: 100 μm. (F) CD4⁺ T cells were isolated from Ctrp4^–/– and WT mice 18 days after EAE induction and the activity phosphorylation of STAT3 and JAK2 was detected by Western blot. (G and H) dLN CD4⁺ T cells isolated from rhCTRP4-treated group or control group were expanded with irradiated autologous-presenting cells plus 10 μg/mL (G) or indicated concentrations (H) of MOG_35–55 peptide for 72 hours and subjected to cell-proliferation assay to determine T cell recall response based on BrdU assay (G) or quantitate the productions of IL-17A and IFN-γ (H). (I) Ctrp4^–/– female mice were immunized with MOG_35–55 peptide to induce EAE; 500 ng/mice rhCTRP4 or control (BSA) was administered intraperitoneally every day starting at disease onset on day 9 after immunization until sacrificed to restore the level of CTRP4. Clinical scores are depicted (n = 8/group). Data are represented as mean ± SEM and are from 1 of 3 independent experiments with similar results. (A and I) Statistical significance was determined using 2-way repeated-measures ANOVA. Data were analyzed by unpaired Student’s t test or Mann-Whitney U test as appropriate after assessing for distribution (C–E, G, and H). *P < 0.05; **P < 0.01; ***P < 0.001.