Decreased cell infiltrates and IFN-γ production of liver mononuclear cells in 2-OA-BSA immunized CD1d^-/- mice.

<p>(A-C) CD1d^-/- and wild type mice were immunized with 2-OA-BSA at 0, 2, 4, 6 and 8 and sacrificed at week 12. (A) Liver total mononuclear cells (MNCs) were measured. (B) The numbers of T (CD3⁺ NK1.1^-), NKT (CD3⁺NK1.1⁺), NK (CD3^-NK1.1⁺) and B (CD19⁺) cells were measured. (C) The numbers of CD4⁺ and CD8⁺ T cells were detected. n = 16–20 per group. *, p<0.05. (D) CD1d^-/- or wild type mice were immunized with 2-OA-BSA at weeks 0, 2, and 4 and sacrificed 3 days after last immunization. IFN-γ production of liver mononuclear cells stimulated with anti-CD3 and anti-CD28 Abs for 2 days was measured by ELISA. n = 8 mice for PBS treated group, n = 13–17 mice for 2-OA-BSA immunized group. *, p<0.05; **, p<0.01.</p

Lower serum levels of IFN-γ after OCH injection.

<p>C57BL/6 mice were intravenously injected with α-GalCer, OCH, or PBS. Serum samples were collected at 2 and 18 hours after α-GalCer, OCH, or PBS injection. IFN-<i>γ</i> (A) and IL-4 (B) were measured by ELISA. n = 10 mice per group. ***, p<0.001.</p

Decreased AMAs in 2-OA-BSA immunized CD1d^-/- mice.

<p>(A) iNKT cell deficiency in CD1d^-/- mice was confirmed by staining with CD3 and CD1d tetramer. (B) CD1d^-/- and wild type mice were immunized with 2-OA-BSA at 0, 2, 4, 6 and 8 and sacrificed at week 12. Serum levels of IgM and IgG to mPDC-E2 were measured by ELISA. n = 10 mice for each group. *, p<0.05.</p

Increased serum AMAs in mice injected with 2-OA-BSA/OCH.

<p>Wild type mice were immunized with 2-OA-BSA and α-GalCer (group name: 2-OA/a-GC), OCH (group name: 2-OA/OCH) or PBS (group name: 2-OA/PBS) at weeks 0, 2, 4, 6 and 8. At week 12, serum levels of autoantibodies to mPDC-E2 were measured by ELISA. n = 9–10 mice per group. *, p < 0.05 in 2-OA/a-GC to 2-OA/PBS; #, p < 0.05 in 2-OA/OCH to 2-OA/PBS.</p

The increase of portal inflammation and fibrosis in mice injected with 2-OA-BSA/OCH.

<p>Mice were immunized with 2-OA-BSA and α-GalCer (group name: 2-OA/a-GC), OCH (group name: 2-OA/OCH) or PBS (group name: 2-OA/PBS) at weeks 0, 2, 4, 6 and 8 and sacrificed at week 12. (A) Representative stained liver sections of haematoxylin and eosin (H&E) and Masson’s trichrome stain. (B) Histopathological scores of individual livers on portal inflammation and fibrosis. 0 = no significant change, 1 = minimal, 2 = mild, 3 = moderate, and 4 = severe pathology. Individual symbols each represent a single mouse.</p

Pathological changes in the liver of IFN-γ^−/− mice immunized with 2OA-BSA.

<p>(A) Representative H&amp;E stained of liver sections from IFN-γ^−/− mice compared with WT mice 8 weeks after 2OA-BSA immunization. Portal inflammatory changes with interlobular bile duct damage (red arrow) were observed in WT mice; normal bile ducts in IFN-γ^−/− mice (blue arrows). (B) Scoring of portal inflammation and bile duct damage in sections of liver from WT and IFN-γ^−/− mice (each group, n = 16).</p

Lymphoma-like T cell infiltrations were only found in homozygous dnTGFβRII p40^−/− and dnTGFβRII mice.

<p>A. Representative immunophenotype of hepatic lymphocytes (upper panels) and H&amp;E stained sections (lower panels) from inbred dnTGFβRII p40^−/− mice with lymphomatous lesions at age of 10 weeks. CD4 and CD8 double negative TCRβ⁺NK1.1⁺ cells are predominant in the liver of dnTGFβRII p40^−/−. Typical diffuse lymphomatous lesions were found in liver and spleen. B. Representative immunophenotype of liver infiltrating lymphocytes (upper panels) and H&amp;E stained sections from inbred dnTGFβRII mice with lymphomatous lesions at age of 12 weeks. C. Relative copy number of dnTGFβRII transgene detected by real-time PCR. D. The percentage of homozygous and hemizygous offspring from hemizygous TGFβRII parents.</p