Early Virus-Host Interactions Dictate the Course of a Persistent Infection
Figure 6
Priming with ArmΔGPC changes cellular tropism during Cl13 infection.
C57Bl6/J mice (n = 4/group) were primed and infected as in Fig. 5 and spleens were harvested at 72 hours post-infection. A) Spleens were incubated with 1 mg/mL collagenase D/100 µg DNaseI and manually dissociated through 100 µm nylon screens. Cells were identified by flow cytometry through staining and gating with the following fluorescently conjugated anti-mouse antibodies: DCS: lin-(CD90.2/CD19), CD11chigh; pDCs: lin-, CD11c+, SiglecH+, PDCA-1+, B220+; Eosinophils: lin-, CD11b+, Ly6G-, Ly6clow, SSC-Ahigh; Neutrophils: lin-, CD11b+, F4/80-, Ly6G+; Macs/Monocytes: lin-, CD11b+, Ly6G-, Ly6c+, F4/80+; B cells: B220+, CD3-, CD8-, NK1.1-; T cells: B220-, NK1.1-, CD3+, CD8+ or CD4+; NK cells: CD3low, B220-, NK1.1+; Endothelial cells: lin-, CD45-, gp38+, CD31+; FRCs: lin-, Cd45-, gp38+, CD31-, CD35low. Representative data from one of two independent experiments are shown with SEM calculated and displayed using GraphPad Prism software. Statistical analysis (unpaired t-test) is shown between mock primed and ArmΔGPC primed Cl13 infected groups. NS, not significant; *p<0.05; **p<0.005; **p<0.0005. B–D) Spleens treated as in Fig. 5 and 6 µm sections were stained with guinea pig serum to LCMV GPC (Green) and antibodies used to visualize mature macrophages (F4/80, red, B and D) and fibroblastic reticular cells (ER-TR7, red, C and D). Yellow arrows indicate areas of costaining. The two examples from each staining are from different mice. D) Spleens of naïve mice are shown for comparison and specificity of LCMV serum. White bars indicate 100 µm. All micrographs are representative examples of micrographs from two independent experiments.