TABLE 2.

Percentages of CD4+ and CD8+ T cells isolated prior to challenge (time zero) from the MLN, PP, IEL, and LP of naive, VP6-immunized, LT(R192G)-immunized, and VP6/LT(R192G)-immunized mice that were stimulated to produce cytokines as determined by FACSa

GroupCell type% CD4+ T cells% CD8+ T cells
IL-17+IFN-γ+IL-17+IFN-γ+IL-5+
NaiveIEL
LP0.30
MLN
PP
VP6/LT(R192G) immunizedIEL2.110.35
LP2.800.160.161.540.28
MLN0.14
PP0.71
VP6 immunizedIEL0.10
LP0.140.130.95
MLN
PP
LT(R192G) immunizedIEL
LP0.160.180.59
MLN
PP
  • a Lymphocytes were isolated and either unstimulated or stimulated with VP6 for 18 h, the last 4 h of culture in the presence of brefeldin A, and then stained with monoclonal antibodies to cell surface markers and intracellular cytokines. Cells were analyzed by gating on forward and side scatter to determine the lymphocyte population and then by gating on CD4+ or CD8+ T cells. The percentage of CD4+ or CD8+ cells staining for CD44high and the cytokine being measured was determined. Isotype control values were ≤0.03 and were subtracted from the cytokine-specific values. The percentage of unstimulated cells staining for each cytokine was subtracted from the percentage of VP6-stimulated cells. −, Values of <0.1%.