Figure 3. 2-DG blocks cytolytic activity in CD8⁺ effector T cells.

A. Effect of 50mM 2-DG on induction of perforin and granzyme C mRNA levels, as assessed by quantitative RT-PCR. Data represent average of two independent assays. B. Cytolytic activity of CD8⁺ effector T cells was determined in a 4-hour ⁵¹Cr-release assay, in the presence or absence of 50 mM 2-DG. Data represent the average of five independent experiments. Inhibition by 2-DG was statistically significant at each E:T ratio (P<0.01). C. Conjugate formation was assessed between 2C and P815-B7.1 targets, in the presence or absence of 50mM 2-DG. Calcein AM-labeled T cells (green) were combined with PKH-labeled P815-B7.1 cells (red) for indicated periods of time. Flow cytometry was used to determine the percentage of conjugates out of the total number of cells. Data represent the average + SD of three pooled experiments. D. MTOC reorientation in T cell/target cell conjugates in the presence or absence of 50mM 2-DG, as measured by tubulin staining (red). Data shown is a representative image of at least 20 conjugates analyzed. E. Formation of the cSMAC in T cell/target cell conjugates. Antibodies against PKCθ (green) and talin (red) were used. Target cells were stained with CMAC (blue). Images are representative of at least 20 conjugates analyzed.