Figure 1. In Vitro T Cell Proliferation and Activation is Augmented in Cmah−/− Mice.

(A and C) CFSE-labeled mouse splenocytes (A) or negatively isolated CD8+ T cells (C) were activated in culture for 3 or 5 days with α-CD3 α-CD28 beads. Proliferation was assayed by visualizing CFSE dilution (left most panels), while CD62L and CD25 expression was used to quantify activation differences (middle and right panels). Red-shaded lines represent either representative Cmah−/− mice (A) or Cmah−/− isolated CD8+ T cells (C), while blue-shaded lines represent either representative WT mice (A) or WT isolated CD8+ T cells (C). (B) CFSE-labeled mouse splenocytes were activated in culture for 5 days with α-CD3 α-CD28 beads. Proliferation indices and division indices on CD8 T cells were calculated based on analysis of CFSE dilution. (D) Mouse splenocytes were activated in culture for 5 hours with α-CD3 α-CD28 beads, and assayed for CD62L (left panel) and CD25 (right panel). (E) Mouse splenocytes were stimulated with PMA for 5 hours and assayed for IFNγ production. Black triangles – WT cells, red squares Cmah−/− cells. *p 0.05, #p<0.1, student’s t test. N=5 mice per group (A, B, D), pooled T cells isolated from 5 mice per group (C), and 4 mice per group (E). Results are representative of at least two experiments.