We quantified the epitope-specific CD8+ T-cell response to RA59-gfp/gp33 in the spleen and in the CNS
after both i.c. and i.p. inoculation. Thus, 4-week-old B6 mice were inoculated either i.c. or i.p. and animals were sacrificed at day 7 postinfection (after i.c. inoculation) and on day 8 postinfection (after i.p. inoculation), the peak of inflammation for each route. Splenocytes were isolated from both sets of mice; CNS mononuclear cells were isolated from only those mice inoculated by the i.c. route. An intracellular IFN-γ assay was used to quantify the percentage of CD8+ T cells specific for the MHV spike protein CD8+ T-cell epitope S598 and for gp33 (19, 25). As shown in Fig. Fig.2,2, S598 and gp33 epitope-specific CD8+ T cells are detected in both the spleen and the CNS. Response in the spleen is similar whether inoculation is carried out either i.c. or i.p.; following i.c. infection, the levels of CD8+ T cells specific for both epitopes are higher in the CNS, the site of viral replication, than in the spleen, as observed previously
We quantified the epitope-specific CD8+ T-cell response to RA59-gfp/gp33 in the spleen and in the CNS after both i.c. and i.p. inoculation. Thus, 4-week-old B6 mice were inoculated either i.c. or i.p. and animals were sacrificed at day 7 postinfection (after i.c. inoculation) and on day 8 postinfection (after i.p. inoculation), the peak of inflammation for each route. Splenocytes were isolated from both sets of mice; CNS mononuclear cells were isolated from only those mice inoculated by the i.c. route. An intracellular IFN-γ assay was used to quantify the percentage of CD8+ T cells specific for the MHV spike protein CD8+ T-cell epitope S598 and for gp33 (19, 25). As shown in Fig. Fig.2,2, S598 and gp33 epitope-specific CD8+ T cells are detected in both the spleen and the CNS. Response in the spleen is similar whether inoculation is carried out either i.c. or i.p.; following i.c. infection, the levels of CD8+ T cells specific for both epitopes are higher in the CNS, the site of viral replication, than in the spleen, as observed previously
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