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Figure 3 | Retrovirology

Figure 3

From: Isolated receptor binding domains of HTLV-1 and HTLV-2 envelopes bind Glut-1 on activated CD4+ and CD8+ T cells

Figure 3

TCR stimulation results in Glut-1 expression and concomitant glucose uptake in CD4 and CD8 lymphocytes: Induction of H1 RBD and H2 RBD binding. CD4+ and CD8+ T lymphocytes were isolated by negative selection and stimulated via the TCR using αCD3/αCD28 mAbs. (A) Non-activated and TCR-activated T cells were used for binding assays with mAb1418 followed by incubation with a FITC-conjugated αmouse IgG. Intracellular Glut-1 levels were monitored in permeabilized cells using the C-term Glut-1 polyclonal antibody followed by incubation with a FITC-conjugated sheep αrabbit IgG antibody. Filled histograms depict binding in the presence of the secondary FITC-conjugated antibody alone. Expression of the HTLV-1 Env receptor was detected by a 30 min incubation of the non-activated and TCR-activated cells with rabbit rFc-tagged H1RBD fusion protein at 37°C and binding was revealed by a 20 min incubation at 4°C with a FITC-conjugated sheep αrabbit IgG antibody. Binding to the H2RBD domain fused directly to EGFP (H2RBD-EGFP) was detected following a 30 min incubation at 37°C. (B) Glucose uptake was assayed by incubating non-activated and TCR-activated CD4 and CD8 T cells (1 × 106) with 2-deoxy-D [1-3H]glucose (2 μCi) for 45 min at 37°C. Uptake for each cell population is expressed as mean counts per minute (CPM) for triplicate samples, error bars indicate SD.

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