Figure 10.

The pathogenetic role of NKT cells in concanavalin A-acute hepatitis. C57BL6 male mice have been pretreated with α-NK1.1 or α-IgG antibody (as a negative control) 200 μg/topo intraperitoneally the day before hepatitis induction. Then hepatitis was induced by the administration of Con A at the concentration of 15 mg/kg intravenously and the mice were sacrificed 24 hours later. (A) Histological evaluation, performed with hematoxylin and eosin staining, of necrosis area on liver sections of mice treated with Con A alone or in combination with α-NK1.1 or α-IgG antibody. Total RNA extracted from liver was used to evaluate by quantitative real-time PCR the relative mRNA expression of (B) proinflammatory genes Cd49b, IFN-γ, IL-6, IL-1β, and TNF-α. The data are normalized to GAPDH mRNA. To determine whether the difference in the severity of the hepatitis developed after the administration of Con A in GPBAR1^+/+and GPBAR1^–/– mice could depend on the absence of the GPBAR1 receptor in NKT cells, we performed a cell transfer experiment of liver-purified NKT cells obtained from GPBAR1^+/+ and GPBAR1^–/– 24 hours after the treatment with Con A 15 mg/kg. Subsequently, we transferred 4 × 10⁶ NKT cells resuspended in a saline solution with Con A, administered at a dose of 5 mg/kg. Data shown are (C, D) plasmatic levels of AST and ALT for each experimental group. (E) Histological evaluation, performed with hematoxylin and eosin staining, of necrosis area on liver sections. Total RNA extracted from liver was used to evaluate by quantitative real-time PCR the relative mRNA expression of (F) proinflammatory genes IFN-γ, IL-6, IL-1β, and TNF-α. Each point represents a single mouse. *P < .05.