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Phenotypic Characterization of Collagen Induced Arthritis in DBA/1J Mice. DBA/1J mice were immunized with bovine type II collagen (N = 11–55; arthritic: n = 6–18; non-arthritic: n = 5–37) or PBS (N = 4–14). A) Representative photographs and E) 3D-rendered micro-CT radiographs of arthritic (red arrow = bone erosion), non-arthritic, and PBS-injected mouse hind limbs. Changes in B) clinical arthritis score, C) anti-bovine type II collagen (CII) IgG and D) anti-mouse CII IgG over time. Dashed line represents the antibody positivity cut-off, indicating the limit of detection. F) Hind limb bone mineral density and G) bone mineral density of bCII-immunized fore and hind limbs over time. H) Pain-like behaviour at day 47 post primary immunization. Graphs show the median [IQR], with each symbol in F-H) representing an individual mouse. For B-D), statistical analysis was performed using a mixed effects model with the Geisser-Greenhouse correction and Tukey's multiple comparisons test, with resulting p-values shown in the graphs. Kruskal-Wallis with Dunn's multiple comparisons test was used for F) and H), and the resulting p-values were p = 0.0008 and p = 0.0003 respectively, with p-values for pairwise comparisons indicated on the graphs. For G), a Spearman correlation coefficient (r) was calculated and p < 0.05 was considered significant.

Journal: Journal of Translational Autoimmunity

Article Title: T cell proliferative response to a homocitrullinated peptide correlates with joint pathology in collagen induced arthritis

doi: 10.1016/j.jtauto.2025.100345

Figure Lengend Snippet: Phenotypic Characterization of Collagen Induced Arthritis in DBA/1J Mice. DBA/1J mice were immunized with bovine type II collagen (N = 11–55; arthritic: n = 6–18; non-arthritic: n = 5–37) or PBS (N = 4–14). A) Representative photographs and E) 3D-rendered micro-CT radiographs of arthritic (red arrow = bone erosion), non-arthritic, and PBS-injected mouse hind limbs. Changes in B) clinical arthritis score, C) anti-bovine type II collagen (CII) IgG and D) anti-mouse CII IgG over time. Dashed line represents the antibody positivity cut-off, indicating the limit of detection. F) Hind limb bone mineral density and G) bone mineral density of bCII-immunized fore and hind limbs over time. H) Pain-like behaviour at day 47 post primary immunization. Graphs show the median [IQR], with each symbol in F-H) representing an individual mouse. For B-D), statistical analysis was performed using a mixed effects model with the Geisser-Greenhouse correction and Tukey's multiple comparisons test, with resulting p-values shown in the graphs. Kruskal-Wallis with Dunn's multiple comparisons test was used for F) and H), and the resulting p-values were p = 0.0008 and p = 0.0003 respectively, with p-values for pairwise comparisons indicated on the graphs. For G), a Spearman correlation coefficient (r) was calculated and p < 0.05 was considered significant.

Article Snippet: Immunization-grade bovine type II collagen (bCII) protein was purchased from Chondrex Inc (20022; USA).

Techniques: Micro-CT, Injection

Splenic T Cell Proliferation Following Stimulation with Type II Collagen Protein. Splenocytes isolated at day 49 post primary immunization from DBA/1J mice immunized with bovine type II collagen (bCII) (arthritic: N = 8; non-arthritic: N = 7) or PBS (N = 10) were cultured in media alone or 80 μg/mL bCII protein. Following 72 h incubation, A) CD4 + and B) CD8 + T cell proliferation following stimulation with bCII was measured by flow cytometry and is shown as a mean (±SD) stimulation index, with data points representing individual mice. Stimulation indices greater than 1.6 (dashed line) were considered a positive response. Statistical analysis was performed using one-way ANOVA with Tukey's multiple comparisons test, and the resulting p-values were p = 0.0054 and p = 0.0987 respectively, with p-values for pairwise comparisons indicated on the graphs. ns: not significant.

Journal: Journal of Translational Autoimmunity

Article Title: T cell proliferative response to a homocitrullinated peptide correlates with joint pathology in collagen induced arthritis

doi: 10.1016/j.jtauto.2025.100345

Figure Lengend Snippet: Splenic T Cell Proliferation Following Stimulation with Type II Collagen Protein. Splenocytes isolated at day 49 post primary immunization from DBA/1J mice immunized with bovine type II collagen (bCII) (arthritic: N = 8; non-arthritic: N = 7) or PBS (N = 10) were cultured in media alone or 80 μg/mL bCII protein. Following 72 h incubation, A) CD4 + and B) CD8 + T cell proliferation following stimulation with bCII was measured by flow cytometry and is shown as a mean (±SD) stimulation index, with data points representing individual mice. Stimulation indices greater than 1.6 (dashed line) were considered a positive response. Statistical analysis was performed using one-way ANOVA with Tukey's multiple comparisons test, and the resulting p-values were p = 0.0054 and p = 0.0987 respectively, with p-values for pairwise comparisons indicated on the graphs. ns: not significant.

Article Snippet: Immunization-grade bovine type II collagen (bCII) protein was purchased from Chondrex Inc (20022; USA).

Techniques: Isolation, Cell Culture, Incubation, Flow Cytometry

RA-Specific Serum IgG Antibodies in DBA/1J Mice. Sera were collected biweekly from DBA/1J mice immunized with bovine type II collagen (bCII) (arthritic: N = 9–18; non-arthritic: N = 17–37), or PBS (N = 14) and tested via indirect ELISA for A) anti-CitP, and B) anti-HomoCitP IgG antibodies. Graphs show the median [IQR] for each group, with a cut-off indicating the limit of detection (dashed line). Statistical analysis was performed using a mixed effects model with the Geisser-Greenhouse correction and Tukey's multiple comparisons test. The resulting p-values are indicated on the graphs.

Journal: Journal of Translational Autoimmunity

Article Title: T cell proliferative response to a homocitrullinated peptide correlates with joint pathology in collagen induced arthritis

doi: 10.1016/j.jtauto.2025.100345

Figure Lengend Snippet: RA-Specific Serum IgG Antibodies in DBA/1J Mice. Sera were collected biweekly from DBA/1J mice immunized with bovine type II collagen (bCII) (arthritic: N = 9–18; non-arthritic: N = 17–37), or PBS (N = 14) and tested via indirect ELISA for A) anti-CitP, and B) anti-HomoCitP IgG antibodies. Graphs show the median [IQR] for each group, with a cut-off indicating the limit of detection (dashed line). Statistical analysis was performed using a mixed effects model with the Geisser-Greenhouse correction and Tukey's multiple comparisons test. The resulting p-values are indicated on the graphs.

Article Snippet: Immunization-grade bovine type II collagen (bCII) protein was purchased from Chondrex Inc (20022; USA).

Techniques: Indirect ELISA