Sustained CD8+ T Cell Memory Inflation after Infection with a Single-Cycle Cytomegalovirus (original) (raw)
Figure 6
Contaminating spread-competent MCMV cannot account for CD8+ T cell memory inflation.
A) MEFs were infected with 1×105 pfu of ΔgL with or without 3 pfu of wild-type K181 MCMV. Shown are representative images of infected cultures taken 4 and 14 days after infection. Data is representative of at least 8 independent experiments. B) BALB/c-SCID mice were infected with the indicated amounts of wild-type K181 MCMV or 1×105 pfu of ΔgL MCMV and sacrificed at the indicated day post infection. Uninfected control samples were derived from naïve C57BL/6 mice. Viral DNA was quantified in the spleens and salivary glands by qPCR. Data was combined from 2 independent experiments. One mouse infected with a single pfu of wild-type virus displayed detectable viral DNA in the salivary gland, but not the spleen (indicated by the asterisks). A correlation was found between the salivary gland and spleen in all other mice. The limit of detection in these assays was estimated to be 30 copies of viral DNA per 500 ng of total DNA as measured by a standard curve of the viral E1 gene cloned into a plasmid. C) B6 mice were infected with 10 or 100 pfu of wild-type K181 MCMV and antigen-specific T cells were tracked in the peripheral blood by tetramer staining. Each box shows the T cells specific for m139, M38 and IE3 within a single mouse. The boxes marked with an asterisk indicate mice in which IE3-specific T cells failed to inflate despite accumulation of m139- and M38-specific T cells. Data was combined from 2 independent experiments.