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(A) Schematic representation of the adoptive CD4⁺CD45RB high T-cell transfer model of chronic colitis. CD4⁺CD45RB high T cells were isolated from BALB/c donor spleens and intravenously <t>transferred</t> <t>into</t> <t>C.B-17</t> scid recipient mice. Beginning after cell transfer, mice received daily subcutaneous administration of CIP-3 (1 mg/kg or 5 mg/kg) or vehicle. Disease progression was monitored for 30 days. Endpoints included disease activity index (DAI), colon length, and serum cytokine analysis. (B) CIP-3 reduced disease severity in a dose-dependent manner. DAI (combined weight loss and stool score) was assessed longitudinally. Vehicle-treated mice developed progressive colitis, whereas CIP-3 treatment significantly attenuated disease progression, with greater efficacy observed at 5 mg/kg. (C) CIP-3 mitigated colonic shortening associated with inflammation. Colon length was measured on day 30. Vehicle-treated mice exhibited marked shortening relative to no-colitis controls, whereas CIP-3 treatment significantly restored colon length in a dose-dependent manner. (D-E) CIP-3 suppressed systemic inflammatory cytokines. Serum TNF-α ( D) and IL-6 (E) levels were quantified by ELISA at study termination. Vehicle-treated mice showed elevated cytokine concentrations, while CIP-3 significantly reduced TNF-α and IL-6 levels, with stronger suppression at 5 mg/kg. Data represent mean ± SEM (n = 8 per group). Statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparison test for endpoint analyses and two-way ANOVA for longitudinal DAI measurements. * P < 0.05, ** P < 0.01, *** P < 0.001 versus colitis + vehicle group.
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(A) Schematic representation of the adoptive CD4⁺CD45RB high T-cell transfer model of chronic colitis. CD4⁺CD45RB high T cells were isolated from BALB/c donor spleens and intravenously transferred into C.B-17 scid recipient mice. Beginning after cell transfer, mice received daily subcutaneous administration of CIP-3 (1 mg/kg or 5 mg/kg) or vehicle. Disease progression was monitored for 30 days. Endpoints included disease activity index (DAI), colon length, and serum cytokine analysis. (B) CIP-3 reduced disease severity in a dose-dependent manner. DAI (combined weight loss and stool score) was assessed longitudinally. Vehicle-treated mice developed progressive colitis, whereas CIP-3 treatment significantly attenuated disease progression, with greater efficacy observed at 5 mg/kg. (C) CIP-3 mitigated colonic shortening associated with inflammation. Colon length was measured on day 30. Vehicle-treated mice exhibited marked shortening relative to no-colitis controls, whereas CIP-3 treatment significantly restored colon length in a dose-dependent manner. (D-E) CIP-3 suppressed systemic inflammatory cytokines. Serum TNF-α ( D) and IL-6 (E) levels were quantified by ELISA at study termination. Vehicle-treated mice showed elevated cytokine concentrations, while CIP-3 significantly reduced TNF-α and IL-6 levels, with stronger suppression at 5 mg/kg. Data represent mean ± SEM (n = 8 per group). Statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparison test for endpoint analyses and two-way ANOVA for longitudinal DAI measurements. * P < 0.05, ** P < 0.01, *** P < 0.001 versus colitis + vehicle group.

Journal: bioRxiv

Article Title: AI-designed cyclic peptides enable controllable modulation of the CD28 immune checkpoint

doi: 10.64898/2026.03.06.710051

Figure Lengend Snippet: (A) Schematic representation of the adoptive CD4⁺CD45RB high T-cell transfer model of chronic colitis. CD4⁺CD45RB high T cells were isolated from BALB/c donor spleens and intravenously transferred into C.B-17 scid recipient mice. Beginning after cell transfer, mice received daily subcutaneous administration of CIP-3 (1 mg/kg or 5 mg/kg) or vehicle. Disease progression was monitored for 30 days. Endpoints included disease activity index (DAI), colon length, and serum cytokine analysis. (B) CIP-3 reduced disease severity in a dose-dependent manner. DAI (combined weight loss and stool score) was assessed longitudinally. Vehicle-treated mice developed progressive colitis, whereas CIP-3 treatment significantly attenuated disease progression, with greater efficacy observed at 5 mg/kg. (C) CIP-3 mitigated colonic shortening associated with inflammation. Colon length was measured on day 30. Vehicle-treated mice exhibited marked shortening relative to no-colitis controls, whereas CIP-3 treatment significantly restored colon length in a dose-dependent manner. (D-E) CIP-3 suppressed systemic inflammatory cytokines. Serum TNF-α ( D) and IL-6 (E) levels were quantified by ELISA at study termination. Vehicle-treated mice showed elevated cytokine concentrations, while CIP-3 significantly reduced TNF-α and IL-6 levels, with stronger suppression at 5 mg/kg. Data represent mean ± SEM (n = 8 per group). Statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparison test for endpoint analyses and two-way ANOVA for longitudinal DAI measurements. * P < 0.05, ** P < 0.01, *** P < 0.001 versus colitis + vehicle group.

Article Snippet: Recipient C.B-17 scid mice (7 weeks old; Taconic Biosciences) received 3 × 10 5 CD4+CD45RB high T cells via intravenous injection (200 μL per mouse).

Techniques: Isolation, Biomarker Discovery, Activity Assay, Enzyme-linked Immunosorbent Assay, Comparison