huTNF(DBA/1) Mouse

Figure 4. ELISA detection of TNF protein expression in the serum of 10-week-old female DBA/1 wild-type mice and huTNF(DBA/1) mice (huTNF)*. *The human TNFα protein ELISA kit used for detection was from Thermo Fisher: Human TNF alpha ELISA Kit, Ultrasensitive (Catalog No.: KHC3014); The mouse TNFα protein ELISA kit used for detection was from MULTI SCIENCES: Mouse TNF-a ELISA Kit (EK282/4). **LPS is an endotoxin that can be used to stimulate the immune response in mice. LPS immunization can exacerbate the severity of arthritis ^[3].

Figure 7. Growth Curves and Clinical Scores. The results show no significant difference in body weight between the model and control groups of huTNF(DBA/1) mice and DBA/1 mice. Clinical scores in the model groups of huTNF(DBA/1) and DBA/1 mice significantly increased, indicating successful CIA model construction. The disease progressed more rapidly and severely in the huTNF(DBA/1) model group compared to the DBA/1 model group. After Adalimumab treatment, the average clinical score of the huTNF-CIA+Adalimumab efficacy group was significantly lower than that of the model group, indicating effective disease control in mice (Note: Efficacy is related to treatment duration and dose).

Figure 8. Histopathology. ① No significant abnormalities were observed in the joint cavities of the control groups of huTNF(DBA/1) and DBA/1 mice, with no significant synovial hyperplasia and intact cartilage structure. (Blue: No significant synovial hyperplasia; Red: Normal joint cavity without deformation or fusion; Green: Intact cartilage surface without erosion or damage). ② The model group of DBA/1 mice showed significant synovial hyperplasia and thickening, with inflammatory cells invading the joint cavity. The synovium formed pannus and bone erosion, with excessive proliferation of synovial cells eroding cartilage and bone, resulting in the disappearance of the joint space. Pannus formation or extensive pannus was observed in multiple areas. ③ The model group of huTNF(DBA/1) mice showed synovial cell hyperplasia, proliferation of fibroblast-like cells and synovial phagocytes, accompanied by infiltration of inflammatory cells such as lymphocytes and plasma cells. The cartilage was affected by synovium, pannus, and inflammatory cells, with cartilage ulcer surfaces covered by connective tissue or fibrocartilage and invaded by new blood vessels. Bone tissue damage and joint cavity deformation were observed. ④ In the huTNF(DBA/1)-CIA+Adalimumab efficacy group, Adalimumab treatment significantly improved synovial hyperplasia and inflammatory infiltration; cartilage showed slight erosion (slightly uneven); joints were normal, with mild local cartilage defects and no pannus formation.

Figure 9. Pathological Score. The results show that pathological scores in the model groups (G2, G4) of huTNF(DBA/1) and DBA/1 mice were significantly higher than those in the control groups, with the disease severity being greater in the huTNF(DBA/1) model group. After Adalimumab treatment, synovial hyperplasia and inflammatory infiltration in arthritic mice were significantly improved, with slight cartilage erosion and mild local cartilage defects, resulting in significantly lower pathological scores.

Figure 10. Arthritis scores of huTNF(DBA/1) mice in blank control and TNFi treatment groups (n=4, male). *These data were provided by Cyagen Biosciences partners.

Figure 11. Infliximab efficacy validation protocol and results (8-week-old, female mice, n=5). *Infliximab is a chimeric anti-human TNF‑α monoclonal antibody that specifically neutralizes human TNF‑α with negligible activity against mouse TNF‑α, and is used for the treatment of autoimmune inflammatory diseases. *These data were provided by Cyagen Biosciences partners.