Figure 2.
Figure 2. Recently activated T cells exhibit an intrinsic defect in forming stable interactions with APCs. (A–D) Divided T cells do not form stable interactions with DCs. (A) Experimental set-up. GFP-expressing OT-I CD8+ T cells were labeled with SNARF and transferred into recipient mice. CFP-expressing DCs pulsed with OVAp were injected in the footpad. At 48 h, recipient mice were subjected to intravital imaging of the draining popliteal lymph node. (B) Representative time-lapse images showing that, in the same lymph node, undivided T cells (yellow) established long-lived interactions (arrowheads) with DCs (cyan), whereas divided T cells (green) failed to do so. Bar, 15 µm. (C) Duration of T cell–DC contacts from a representative video is graphed as a function of the GFP/SNARF ratio. Note that undivided T cells (low GFP/SNARF fluorescence ratio) formed interactions >10 min. Results are representative of three independent experiments. (D) The frequency of undivided or divided T cells forming interactions >5 min with DCs (stable contacts) is shown. Results are compiled from seven videos obtained in two independent experiments. (E–H) Recently activated T cells do not form stable interactions with DCs. (E) Experimental set-up. OT-I CD8+ T cells expressing a YFP reporter for IFN-γ (Yeti) were labeled with SNARF and transferred into recipient mice. GFP-expressing DCs pulsed with OVAp were injected in the footpad. At 24 h, recipient mice were subjected to intravital imaging of the draining popliteal lymph node. (F) Representative time-lapse images showing that, in the same lymph node, naive (red) but not activated (orange) T cells established long-lived interactions (arrowheads) with DCs (green). Bar, 15 µm. (G) Duration of T cell–DC contacts from a representative video is graphed as a function of the YFP/SNARF ratio. Note that only unactivated T cells (low YFP/SNARF fluorescence ratio) formed interactions >12 min. Representative of three independent experiments. (H) The frequency of unactivated (naive) or activated T cells forming interactions >5 min with DCs (stable contacts) is shown. Results are compiled from three videos obtained in a representative experiment out of three.

Recently activated T cells exhibit an intrinsic defect in forming stable interactions with APCs. (A–D) Divided T cells do not form stable interactions with DCs. (A) Experimental set-up. GFP-expressing OT-I CD8+ T cells were labeled with SNARF and transferred into recipient mice. CFP-expressing DCs pulsed with OVAp were injected in the footpad. At 48 h, recipient mice were subjected to intravital imaging of the draining popliteal lymph node. (B) Representative time-lapse images showing that, in the same lymph node, undivided T cells (yellow) established long-lived interactions (arrowheads) with DCs (cyan), whereas divided T cells (green) failed to do so. Bar, 15 µm. (C) Duration of T cell–DC contacts from a representative video is graphed as a function of the GFP/SNARF ratio. Note that undivided T cells (low GFP/SNARF fluorescence ratio) formed interactions >10 min. Results are representative of three independent experiments. (D) The frequency of undivided or divided T cells forming interactions >5 min with DCs (stable contacts) is shown. Results are compiled from seven videos obtained in two independent experiments. (E–H) Recently activated T cells do not form stable interactions with DCs. (E) Experimental set-up. OT-I CD8+ T cells expressing a YFP reporter for IFN-γ (Yeti) were labeled with SNARF and transferred into recipient mice. GFP-expressing DCs pulsed with OVAp were injected in the footpad. At 24 h, recipient mice were subjected to intravital imaging of the draining popliteal lymph node. (F) Representative time-lapse images showing that, in the same lymph node, naive (red) but not activated (orange) T cells established long-lived interactions (arrowheads) with DCs (green). Bar, 15 µm. (G) Duration of T cell–DC contacts from a representative video is graphed as a function of the YFP/SNARF ratio. Note that only unactivated T cells (low YFP/SNARF fluorescence ratio) formed interactions >12 min. Representative of three independent experiments. (H) The frequency of unactivated (naive) or activated T cells forming interactions >5 min with DCs (stable contacts) is shown. Results are compiled from three videos obtained in a representative experiment out of three.

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