Antigen-Specific Gene Therapy after Immunisation Reduces the Severity of Collagen-Induced Arthritis
Figure 3
Clinical and histopathological development of arthritis, serum levels of anti-CII IgG, and T cell response after i.v. injection of lentiviral particles at day 26 after CII immunisation. (a) Design of experiments. Immunisation at day 0, lentiviral injection with LNT-GFP or LNT-Ii-CII at day 26 after CII immunisation, booster at day 21, and termination of experiments at days 54–63. (b) The clinical frequency of arthritis. (c) The clinical severity of arthritis (LNT-GFP days 0–43 , days 45–54 , days 56–63 , LNT-Ii-CII days 0–54 , and days 56–63 ). (d) The severity of histopathological synovitis and bone-cartilage erosivity. (e) Histopathological findings in the knee joints at day 56 after CII immunization in a mouse injected with LNT-GFP (top panel) that shows synovitis and bone and cartilage destruction grade 2. The bottom panel shows the knee joint from an LNT-Ii-CII mouse with a nonarthritic knee joint at the same time point. S = synovia, C = cartilage, B = bone, M = meniscus, and bone and cartilage destruction are marked with arrows. Scale bar, 100 μm. (f) Serum levels of CII-specific IgG antibodies at days 45 and 56. (g) Suppressive capacity of T cells measured as levels of IFN-γ in supernatants from cocultures of T cell subsets and CD11c+ cells. The two first bars represent supernatants from cocultures containing CD4+CD25− and CD11c+ cells, the following two CD4+CD25+ cells and CD11c+ cells, and finally 4 bars represent a mix of CD4+CD25− and CD4+CD25+ in a ratio of 1 : 1 or 1 : 10 cocultured with CD11c+ cells. Closed circles represent LNT-GFP and open circles represent LNT-Ii-CII treated mice. In Figure 3(b) the value is calculated using logistic regression, in Figure 3(c) the value is calculated using linear regression, and in Figures 3(d) and 3(f) the values are calculated using Mann Whitney U test. In Figure 3(g) black bars represent cells from LNT-GFP treated mice and white bars represent cells from LNT-Ii-CII treated mice. The value is calculated using two-way ANOVA comparing the increasing IFN-γ levels from T cell cocultures from LNT-GFP mice in ratios from 1 : 1 to 1 : 10 with respect to IFN-γ levels from cells from LNT-Ii-CII mice in ratios from 1 : 1 to 1 : 10. and .
