Functional and Phenotypic Characteristics of Salmonella-Specific Memory T Cells
B10.D2(R101) mice were i.p. immunized with S. typhimurium avirulent strain 247, and 3 weeks post-immunization total spleen cells of immunized mice were stimulated in vitro with increased doses of heat-inactivated S. typhimurium virulent strain IE 147 for 72 h. Total spleen cells of intact (non-immunized) B10.D2(R101) mice were used as the background control.
(A) Analysis of the salmonella-induced in vitro proliferative response.
(B–D) Flow cytometry analyses of lymphocytes 72 h post in vitro stimulation. (B) Cells were gated as follows: leukocytes were gated based on SSC-A vs. FSC-A, and then singlets were gated based on FSC-H vs. FSC-A, and dead cells were excluded by staining with PI. (C) The relative number (%) of T cells (CD3+) in the population of live singlet leukocytes. (D) CD4/CD8 T cells ratio in the population of CD3+ live singlet leukocytes.
Data are representative of 2 independent experiments and shown as mean ± SD (n = 3). ∗p < 0.03, ∗∗p < 0.01, ∗∗∗p < 0.001 (unpaired Student's t-test). NS, not significant. See also Figures S1 and S4.