Fig. 7: LCMV-Docile S-segment contributes to slow propagation thereby promoting chronic infection.
From: Slow viral propagation during initial phase of infection leads to viral persistence in mice
a Schematic of chimeric virus S(WE)/L(Clone 13) and S (Docile)/L(Clone 13) is shown. b BHK-21 cells were infected with the chimeric virus S(WE)/L(Clone 13) or S (Docile)/L(Clone 13) at MOI 0.5. At the indicated timepoints post-infection, monensin was added. Sixteen hours later, LCMV infected cells were quantified by anti-LCMV-NP staining (n = 5). c, d BHK-21 cells were infected with chimeric virus S(WE)/L(Clone 13) or S (Docile)/L(Clone 13) at MOI 0.01 (c), MOI 1 (d). At 24 h post-infection, BHK-21 cellular RNA was isolated and GP RNA, S-IGR RNA, and NP RNA were quantified by RT-PCR (n = 6). e GM-CSF induced BMDCs were infected with chimeric virus S(WE)/L(Clone 13) or S (Docile)/L(Clone 13) or WE WT virus at MOI 1. Twenty four hours post-infection, IFN-α levels were determined in cell supernatants (n = 6). f A schematic of chimeric virus S(WE-GP-Docile-NP)/L(Clone 13) and S (Docile-GP-WE-NP)/L(Clone 13) is shown. g–m C57BL/6 mice were infected with 2 × 105 pfu of chimeric virus S(WE)/L(Clone 13), S (Docile)/L(Clone 13), S(WE-GP-Docile-NP)/L(Clone 13), and S (Docile-GP-WE-NP)/L(Clone 13). g At day 1 and day 2 post-infection, serum IFN-α levels were determined (n = 6). h Numbers of tet-gp33+ CD8+ T cells were determined in blood (left panel), and spleen tissue (right panel) 7 days post-infection (n = 6–10). i Frequency of short-lived effector cells (SLEC, KLRG1+ IL-7R-) were quantified in blood (left panel), and spleen (right panel) from LCMV-specific CD8+ T cells 7 days post-infection (n = 6–10). j Expression of T cell surface molecules from spleen tet-gp33+ CD8+ T cells is shown (representative blots of n = 6–10 are shown, dotted line represents surface molecule expression of CD19+ cells from S(WE)/L(Clone 13) infected hosts). k Blood cells, and l single cell suspended splenocytes were re-stimulated with LCMV-specific CD8+ T cell epitopes as indicated or left untreated (negative control: n.c.) followed by staining for IFN-γ and TNF-α (n = 6). m Seven days post-infection, virus titers were determined in the spleen, liver, lung, and kidney tissue (n = 6–10). (Error bars show SEM, *p < 0.05, **p < 0.01, ***p < 0.001, and ns indicates statistically not significant between the indicated groups).
