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. Author manuscript; available in PMC: 2014 Sep 1.
Published in final edited form as: J Immunol. 2013 Aug 5;191(5):2372–2383. doi: 10.4049/jimmunol.1300107

Figure 6. The SKAP55 binding region in ADAP regulates stable contacts and late stage DC confinement.

Figure 6

Primary ADAP−/− DO11.10 hCAR T cells were transduced with control adenovirus expressing Thy1.1 or adenovirus expressing Thy1.1 and either wild-type ADAP or the ADAPΔ338 mutant as described in Methods. (A) ADAP−/− DO11.10 T cells transduced with control Thy1.1 adenovirus were labeled with CTO (red) and co-transferred with ADAP−/− DO11.10 T cells transduced with adenovirus expressing Thy1.1 and wild-type ADAP and labeled with CTV (blue) into mice primed with CFSE/IFA with or without OVA antigen (Ag). Graphs show average velocity and T-DC contact time from 3 independent movies at four hours post T cell transfer. (B) Three-dimensional images and corresponding kymograph of a single DC interacting with transferred T cells at 4 hours after transfer. Colored spots match cell images to contacts on the kymograph. (C) CTO-labeled ADAP−/− DO11.10 T cells transduced with ADAPΔ338 adenovirus (red) were co-transferred as in (A) with CTV-labeled ADAP−/− DO11.10 T cells transduced with wild-type ADAP adenovirus (blue). Graphs show average velocity and T-DC contact time from 3 independent movies at 4 hours (top) and 24 hours (bottom) post T cell transfer. (D) Pooled post-T cell-DC contact endpoint confinement ratio from 3 independent movies at 24 hours post-transfer for ADAP−/− DO11.10 T cells transduced with wild-type ADAP (blue) and ADAP−/− DO11.10 T cells transduced with ADAPΔ338 adenovirus (red). *, P<0.05 unpaired t test.