Figure 3
Activation of TLR3 and regulation of
Icam-1,
Vcam-1, and
Cxcl9.
(A–E) Mice were treated with poly(I:C). After 24 hours, RT-PCR analysis was performed in livers from C57BL/6 mice (n = 3) for Icam-1, Vcam-1, and Cxcl9 (A); Cxcl9 in livers from BALB/c and Tlr3–/– mice and Tlr3–/–→BALB/c and BALB/c→Tlr3–/– bone marrow chimeras (B); Cxcl9 (C) or Vcam-1 (D) in livers from Ifnar–/–, Ifngr–/–, and Tnfr1–/– mice plus corresponding wild-type controls (n = 3). (E) Eight hours after poly(I:C) treatment, blood and spleen cells of C57BL/6 mice were analyzed for surface expression of CXCR3 by flow cytometry. Histogram plots show CD8+ T cells gated for low CD44 expression (naive CD8+ T cells, gray shaded area) or high CD44 expression (memory CD8+ T cells, black line; 15%–20% of all CD8+ T cells). (F–J) 107 splenocytes from LCMV-gp33/H-2Db–specific TCR-Tg 318 mice were injected i.v. into C57BL/6 mice (F–I) or Alb-1 mice (J) on day –1. Recipients were then immunized with gp33 (1 mg in PBS) together with CpG on days 0 and 4. One group of mice was additionally treated with poly(I:C) on day 7. On day 8, livers were analyzed for Cxcl9 by RT-PCR (F) or VCAM-1 by histology (magnification, ×200) (G). Ten hours after poly(I:C) treatment, mice were analyzed for expression of CXCR3 on tet-gp33+ CD8+ splenocytes (H) and for absolute numbers of tet-gp33+ CD8+ T cells in the blood (I). (J) Livers of immunized Alb-1 mice were analyzed for the presence of autoreactive T cells using anti-CD90.1/Thy1.1 antibody (magnification, ×50, ×200). *P < 0.05.
