Type I Interferon Receptor Deficiency in Dendritic Cells Facilitates Systemic Murine Norovirus Persistence Despite Enhanced Adaptive Immunity
Fig 8
The adaptive immune response generated in persistently infected CD11c-Ifnar1-/- mice is functional in an Ifnar1-sufficient environment.
(A) CD11c-Ifnar1-/- mice were infected with CW3 (filled gray histograms) or CW3 engineered to express the SIINFELK peptide (Red-lined histograms). Three days after inoculation, splenocytes were isolated and stained with an antibody that recognizes H2-Kb with SIINFEKL bound in the peptide-binding groove. Overlain histograms from individual mice show fluorescence intensity of staining on T cells (CD3+, CD19-), B cells (CD19+, CD3-), Macrophages (CD3-, CD19-, F4/80+, CD11blow), or DCs (CD3-, CD19-, CD11c+, MHCII+) as indicated. (B) A portion of the MNoV VP1 gene was amplified by PCR from spleen tissue 21 days after inoculation and sequenced by Sanger sequencing. The shown chromatogram includes the region encoding the immunodominant CD8 T cell epitope and is representative of sequencing results from four mice. (C and D) Splenocytes were collected from CD11c-Ifnar1-/- and littermate controls 21 days after infection with CW3. 107 splenocytes were injected I.P. into Rag-/- mice that had been infected with CW3 21 days prior. Spleens and Ileums were collected from Rag-/- seven days after splenocyte transfer. Viral titers in tissues were determined by qPCR for viral genomes (C) and plaque assay (D). Data in C and D is combined from two experiments with individual mice shown. Statistical significance was determined by one-way ANOVA (A) or Kruskal-Wallis test (B) or. n.s = p>0.05, * = p≤0.05, ** = p≤0.01, *** = p≤0.001, **** = p≤0.0001.