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Figure 5 | Retrovirology

Figure 5

From: Activation and detection of HTLV-I Tax-specific CTLs by Epitope expressing Single-Chain Trimers of MHC Class I in a rat model

Figure 5

Detection of Tax-specific CTLs by SCTs fused with EGFP. (A) The 293T cells transfected with pEF/RT1AlSCNLEnv371L-EGFP or pEF/RT1AlSCTax180L-EGFP were incubated with 4O1/C8 or control G14 cells. After 1 hour of mixed culture, cells were stained with PE-conjugated anti-rat CD8 antibody and EGFP expression on CD8+ cells were assessed by flow cytometric analysis. (B) Cells in the mixed culture of 4O1/C8 and EGFP-expressing 293T cells were attached on slide glasses by centrifugation, fixed with 4% paraformaldehyde for 15 min at room temperature and then stained with an anti-rat CD8 antibody in combination with a Cy3-conjugated goat anti-mouse IgG (H+L) antibody. Fluorescence and differential interference contrast (DIC) images were obtained with a confocal microscope system and a pair of GFP and CD8 images was overlaid (merge). Arrowheads indicate SCT-EGFP in 4O1/C8 cells. Arrows indicate co-localization of SCT-EGFP and CD8 at the contact site. Similar results were obtained in two independent experiments.

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