Journal: Cellular and Molecular Immunology
Article Title: CD205-TLR9-IL-12 axis contributes to CpG-induced oversensitive liver injury in HBsAg transgenic mice by promoting the interaction of NKT cells with Kupffer cells
doi: 10.1038/cmi.2015.111
Figure Lengend Snippet: NKT cell-mediated liver injury is Kupffer cell- and IL-12-dependent in HBs-Tg mice after CpG injection. ( a , b ) Depletion of Kupffer cells was achieved by pretreatment with GdCl 3 24 h before CpG injection in HBs-Tg mice. Endogenous IL-12 was neutralized by pretreatment with anti-IL-12 mAb 12 h before CpG injection in HBs-Tg mice. Serum ALT levels were determined 24 h after CpG injection ( a ), and the expression levels of FasL and CD69 on hepatic NKT cells were determined by FACS analysis ( b ). ( c ) Kupffer cells were isolated from ODN control or CpG-ODN-treated HBs-Tg mice. After 48 h of culture, the supernatants were collected for IL-12 measurement by ELISA. ( d ) HBs-Tg mice were pretreated with GdCl 3 or saline. Serum IL-12 levels were determined 3 h after CpG injection. All values are shown as the mean±s.e.m. from 6–8 mice per group and are representative of two experiments. * P <0.05 versus control group by Student’s t -test. ALT, alanine aminotransferase; ELISA, enzyme-linked immunosorbent assay; FACS, fluorescence-activated cell sorting; HBs-Tg, hepatitis B surface antigen transgenic; IL, interleukin; mAb, antibody; NKT, natural killer T; ODN, oligodeoxynucleotide.
Article Snippet: Mouse CpG-ODN (HC4033: 5′-TCCATGACGTTCCTGATGCT-3′) and non-CpG-ODN control (HC4034: 5′-GCTTGATGACTCAGCCGGAA-3′) were purchased from HyCult Biotechnology (Uden, Netherlands) and were dissolved in pyrogen-free saline.
Techniques: Injection, Expressing, Isolation, Enzyme-linked Immunosorbent Assay, Fluorescence, FACS, Transgenic Assay