Journal: PLoS ONE

Article Title: Upregulation of sphingosine-1-phosphate receptor 3 on fibroblast-like synoviocytes is associated with the development of collagen-induced arthritis via increased interleukin-6 production

doi: 10.1371/journal.pone.0218090

Figure Lengend Snippet: (A) Development of collagen-induced arthritis in WT and S1P 3 knockout (S1P 3 -KO) mice. Arthritis was evaluated three times per week during Days 21–42. Disease severity was measured by assessing the clinical score of all four paws, in which the maximum possible score for an individual mouse was 16. Statistical analysis was performed at Days 36, 38, 40, and 42. Results are presented as the mean ± SEM (n = 20 mice per group) and were analyzed by t -test with Bonferroni’s correction. * P < 0.05 and ** P < 0.01 (WT vs. KO mice). (B) Synovial inflammation, bone erosion, and cartilage destruction were examined histologically on H&E-stained sections of ankle joints and assigned scores of 1–4. Results are presented as box-and-whisker plots (n = 20 mice per group) and were analyzed by Mann-Whitney U-test. The middle hash, the box, and the whiskers represent the median, interquartile range (IQR), and 10/90 percentile values respectively. * P < 0.05, NS = not significant. (C) Representative images of H&E-stained ankle joint sections are shown at a magnification of 40×. Left panel: WT specimen assigned a score of 3 for all parameters. Right panel: KO specimen assigned a score of 1 for inflammation and 0 for the other parameters. (D) Anti-CII IgG 1 and IgG 2 antibodies in serum from CIA mice were measured in an ELISA. Results are presented as box-and-whisker plots (n = 10 mice per group) and were analyzed by Mann-Whitney U-test. The middle hash, the box, and the whiskers represent the median, IQR, and minimum/maximum values respectively. (E) Expression of TNFα and IL-6 in arthritic synovium was analyzed by real-time RT-PCR. Results are presented as the mean ± SEM (n = 6 mice per group) and were analyzed by t -test. * P < 0.05, NS = not significant.

Article Snippet: Levels of anti-CII IgG 1 and IgG 2 were measured using a Mouse Anti-type II Collagen IgG Subtype Antibody ELISA Kit (Chondrex), according to the manufacturer’s instructions.

Techniques: Knock-Out, Staining, Whisker Assay, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

Journal: International Journal of Molecular Medicine

Article Title: Humulus japonicus extract ameliorates collagen-induced arthritis in mice through regulation of overall articular inflammation

doi: 10.3892/ijmm.2019.4417

Figure Lengend Snippet: Effect of Humulus japonicas (HJ) on the production of anti-type II collagen antibody in plasma. Levels of (A) anti-type II collagen total IgG and its subtypes (B) IgG1, and (C) IgG2a were measured by ELISA in plasma obtained on day 35 from each mice group. Gene expression levels of (D) Il-12rβ2 , (E) Ccr2 , (F) Ccr5 , (G) Ccr3 , (H) Ccr4 , (I) IL-2 and (J) IL-13 were analyzed by RT-qPCR in the paw of CIA mice on day 35 following the first immunization. The vehicle group was set to a value of 1, and average fold-change is shown. Grouped quantitative data are presented as means ± SEM (vehicle group; n=8, HJ group; n=6). Significance was measured using two-tailed Student's t-test. * P<0.05, ** P<0.01, *** P<0.001.

Article Snippet: Plasma samples were collected at the end of the experiment (day 35) for the determination of IgG, IgG1, and IgG2a antibody levels with three commercially available test kits, mouse anti-mouse Type II collagen IgG TMB (2036T, Chondrex), IgG1 TMB (20361T, Chondrex), and IgG2a (20362T, Chondrex) antibody subtype assay kit TMB, according to the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Berberine Delays Onset of Collagen-Induced Arthritis through T Cell Suppression

doi: 10.3390/ijms22073522

Figure Lengend Snippet: The effect of berberine on circulating anti-bovine type II collagen (CII) IgG in the CIA model. ( A ) Anti-CII IgG1, IgG2a, and total IgG at day 28 among all mice (arthritic and non-arthritic) within BBR, PBS (vehicle control), CIA (no treatment control), and non-CIA control animals ( n = 10 per group). ( B ) Anti-CII IgG levels at day 28 compared among arthritic mice only (BBR n = 5; PBS n = 9; CIA n = 9). Statistical comparisons made with the Kruskal–Wallis test with Dunn’s multiple comparisons. ( C ) Anti-CII IgG levels at day 28 compared among BBR-treated mice who developed arthritis (“arthritic”) vs. those that did not (“non-arthritic”). Statistical comparisons made with the Mann–Whitney U test. For all statistical tests in A–C, * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Serum concentrations of anti-collagen type II (anti-CII) total IgG (catalog # 1012T), anti-CII IgG1 (catalog # 20321T), and anti-CII IgG2a (catalog # 20322T) were measured by ELISA (Chondrex, Redmond, WA, USA) according to the manufacturer’s instructions [ , ].

Techniques: MANN-WHITNEY

Journal: International Journal of Molecular Medicine

Article Title: Humulus japonicus extract ameliorates collagen-induced arthritis in mice through regulation of overall articular inflammation

doi: 10.3892/ijmm.2019.4417

Figure Lengend Snippet: Effect of Humulus japonicas (HJ) on the production of anti-type II collagen antibody in plasma. Levels of (A) anti-type II collagen total IgG and its subtypes (B) IgG1, and (C) IgG2a were measured by ELISA in plasma obtained on day 35 from each mice group. Gene expression levels of (D) Il-12rβ2 , (E) Ccr2 , (F) Ccr5 , (G) Ccr3 , (H) Ccr4 , (I) IL-2 and (J) IL-13 were analyzed by RT-qPCR in the paw of CIA mice on day 35 following the first immunization. The vehicle group was set to a value of 1, and average fold-change is shown. Grouped quantitative data are presented as means ± SEM (vehicle group; n=8, HJ group; n=6). Significance was measured using two-tailed Student's t-test. * P<0.05, ** P<0.01, *** P<0.001.

Article Snippet: Plasma samples were collected at the end of the experiment (day 35) for the determination of IgG, IgG1, and IgG2a antibody levels with three commercially available test kits, mouse anti-mouse Type II collagen IgG TMB (2036T, Chondrex), IgG1 TMB (20361T, Chondrex), and IgG2a (20362T, Chondrex) antibody subtype assay kit TMB, according to the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Two Tailed Test