A humanized HLA-DR4 mouse model for autoimmune myocarditis

Figures

• 

  Fig. 1. Effects of hCAM immunization on immunological measurements, body weight and myocarditis. (A) Change in thymidine incorporation compared to medium alone controls. Adding hCAM increased spleen cell proliferation after hCAM immunization significantly more than after control immunization (p = 0.022 for 0.2 μg/ml, p = 0.000061 for 2.0 μg/ml and p = 0.0081 for 20 μg/ml hCAM concentration; n = 5). Responses to hCAM were similar to the global T-cell stimulator, ConA (2 μg/ml). (B) Serum IgG1 levels were increased significantly in hCAM compared to control immunized mice (pmax = 0.0008; n = 5). (C) Serum IgG2c levels were also significantly increased as compared to control immunized mice (pmax = 0.02; n = 5). (D) Mice immunized with hCAM group gained no weight, while vehicle-immunized mice gained 2.4 g and were therefore significantly heavier than hCAM-immunized mice after 3 weeks (p = 0.04; n = 5). (E) Haematoxylin and eosin stained histological sections from hearts of the control immunized mice (panel a) were all clear (0/5), while 5/5 of the hearts (p = 0.0079, Fisher's exact test) from hCAM-immunized mice (panel b) were infiltrated with immune cells (blue areas due to concentrated cell…

• 

  Fig. 2. Effect of hCAM-derived peptide pre-treatment. (A) Basal spleen cell proliferation was increased by pool(1) peptide (n = 15) compared to vehicle treatment (n = 15, p = 0.0045) but not with pool(2) (n = 11). Addition of hCAM (20 μg) stimulated splenocyte proliferation to a similar extent after treatment with vehicle, pool(1) or pool(2) peptides. (B) Serum anti-hCAM IgG1 levels were decreased significantly after treatment with pool(2) peptides (n = 11), compared to the vehicle treated group (n = 15), at all dilutions (pmax = 0.036), while no effect was recorded with pool(1) peptides (n = 15). (C) Serum anti-hCAM IgG2c levels were also decreased significantly with only pool(2) peptides at 1/4000 dilution (p = 0.0014; n = 11 vs 15; Wilcoxon test was applied due to non-Gaussian distribution). (D) The area of sections occupied by leukocyte infiltration and myocyte damage was reduced significantly by pool(2) treatment (17.5 vs 29.9%; p = 0.021; n = 11), whereas pool(1) treatment (n = 15) had no significant effect compared to vehicle treated controls (n = 15). (E) Both peptide treatments significantly increased EF (p = 0.011, n = 10 for pool(2); p = 0.041, n = 13 for pool(1)) and FS (p =…