Unexpected Phenotype of Mice Lacking Shcbp1, a Protein Induced during T Cell Proliferation
Figure 6
Loss of Shcbp1 affects disease severity in CD4+ T cell driven autoimmune disease.
(A) Shcbp1 mRNA expression in mononuclear cells isolated from the brain and spinal cords of healthy controls or from mice immunized with MOG(35–55) on day 28 after disease induction (Normalized to HPRT and healthy controls) (n = 3 mice of each condition, pooled). (B) Shcbp1 expression assessed by immunohistochemistry in spinal cords isolated from healthy controls or Shcbp1+/+ and Shcbp1−/− mice on day 28 after immunization for EAE (n = 3 mice of each condition). (C) Left, mean clinical scores of EAE in male Shcbp1+/+ and Shcbp1−/− mice. Middle, percent survival of male Shcbp1+/+ and Shcbp1−/− mice. Right, disease index (area under the curve) plotted for each mouse individually. Compiled data with n = 18–19 mice of Shcbp1+/+ and Shcbp1−/− genotype, respectively. (D) Left, mean clinical scores of EAE in female Shcbp1+/+ and Shcbp1−/− mice. Right, percent survival of female Shcbp1+/+ and Shcbp1−/− mice (n = 17, 12 Shcbp1+/+ and Shcbp1−/− mice respectively). (E) Left, absolute numbers of total cells, CD4+ T cells, B cells, and macrophages isolated from the spinal cords and brain of Shcbp1+/+ and Shcbp1−/− mice on day 28 after immunization (n = 3 mice of each genotype). (F) Immunohistochemistry for CD3 and H&E staining on sacral spinal cord sections from Shcbp1+/+ and Shcbp1−/− mice on day 28 after immunization with EAE (representative of n = 7 Shcbp1+/+ and n = 6 Shcbp1−/− mice, respectively). (G) Quantification of number of CD3+ cells from the sacral spinal cord sections (n = 7 and n = 6 for Shcbp1+/+ and Shcbp1−/− mice, respectively, p = 0.02). (H) Relative expression of ShcA, Shcbp1, and IL2 in Shcbp1+/+ T cells (left) or Shcbp1−/− T cells (right) after activation with anti-CD3/anti-CD28. Normalized to HPRT and unstimulated cells (n = 2–3 experiments). (I) Upregulation of Shcbp1 in CD4+ T cells stimulated with IL2 (normalized to HPRT and unstimulated cells) (n = 3, p = 0.01). (J) Immunoblotting for Shcbp1 and actin (loading control) in naïve and TH17 skewed cells from Shcbp1+/+ and Shcbp1−/− mice (representative of n = 2 experiments).