Figure 2

Detecting perturbations in the memory T cell pool using an altered peptide approach. NP_324-332K^b+ and altered peptide amino acid sequences are shown in A. Spleen cells from mice infected with Sendai virus 1 month earlier were stimulated with 1μg of either NP_324-332K^b (B, top left) or 5HNP_324-332K^b (B, bottom left) peptide in the presence of BFA and IL-2 or were stained with NP_324-332K^b+ or 5HNP_324-332K^b+ tetramers, CD8 and CD44. Flow cytometric plots show intracellular IFNγ production (B, left) or frequency of tetramer (B, right) in CD8 gated T cells. C, Peripheral blood from mice with normal (1 month post infection) (left) and TCE (12–24 month post infection, mouse #s 13 and 197) (middle and right) memory CD8⁺ T cell pools. Flow cytometric plots show frequency of NP_324-332K^b+- (top) and 5HNP_324-332K^b+- (bottom) specific cells among CD8⁺ T cells. Numbers in the gate indicate frequency of cells among CD8⁺ T cells, numbers outside the gate represent frequency of cells relative to frequency of NP_324-332K^b+ cells. D, Data show frequency of 5HNP_324-332K^b+ relative to NP_324-332K^b+ cells from 1 month memory mice, (symbols indicate individual mice and bar represents mean value) and of 6 individual memory mice 12–24 months post infection exhibiting TCE (>20% NP_324-332K^b+cells). * Represents range % of CD8⁺ T cells binding to tetramer relative to NP_324-332K^b+ based on blood samples taken from 200 Sendai virus infected mice.