FIGURE 4.

Fut7 and functional selectin ligand expression in activated CD4 T cells in SN1 Tg mice. A, CD4 T cells were isolated, activated with plate-bound anti-CD3/CD28 without any added cytokine (none), with IL-12, or with TGFβ1, and subsequently cultured with IL-2 alone, IL-12 plus IL-2, or TGFβ1 plus IL-2 and analyzed by flow cytometry with selectin chimeras every 2 days. Depicted are the fractions that are positive for E-selectin ligands (top row) and P-selectin ligands (bottom row) for each genotype at each time point. Mean values ± S.D. from three experiments. B, qRT-PCR of Fut7 mRNA levels in activated CD4 T cells cultured with cytokines as above. Cells were harvested on day 6. As for neutrophils, values were normalized first to hprt levels. These values were then normalized to WT IL-2 only (=1) and expressed as -fold difference over WT IL-2 only. One of two similar experiments is shown. C, induction of E- and P-selectin ligands on splenic CD4 T cells in response to T. gondii infection. Spleens were harvested 7 days after intraperitoneal injection of T. gondii cysts. Plots are gated on CD4+ cells (gating not shown). Representative FACS plots of WT cells show that E- and P-selectin ligand-expressing cells are largely confined to the CD44-hi (activated) population. D, the fraction of CD44-hi CD4+ cells expressing ligands for either E- or P-selectin was calculated. Each symbol represents an individual mouse. E, total CD4+ PEC recovered on day 7 from each of the 4 genotypes of mice. For D and E: *, different (p < 0.01) from all other groups; **, no difference between groups.