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. Author manuscript; available in PMC: 2017 Jul 15.
Published in final edited form as: J Immunol. 2016 Jun 6;197(2):644–654. doi: 10.4049/jimmunol.1600178

Figure 1. Superior JR209 TEM cytotoxic function compared with TCM is due to increased serial killing by TEM and a greater likelihood of induction of effector molecule polarization.

Figure 1

(A) Analysis of TCM and TEM phenotypic markers CD44, CD25 and CD62L by flow cytometry. (B) Cytotoxic properties of JR209 TCM (dashed) and TEM (solid) toward T2-A2Kb APCs loaded with 10 μM gp100209-217(2M) at effector:target ratios indicated (Left). Plot is representative of triplicate experiments. Specific lysis of TCM and TEM toward T2-A2Kb APCs loaded with 10 μMgp100209-217(2M) at 2:1 effector:target ratio (Right). Specific lysis calculated as described in methods (n=6, Two tailed t-test – p < 0.0001). (C) Expression of Fas ligand (FasL), granzyme B (GrnB), and perforin (Perf) by JR209 TCM (light) and TEM (dark) was determined by RT-PCR and presented as expression relative to naïve JR209 T cells (n = 3, Two tailed t-test – FasL p = 0.38, GrnB p = 0.002, Perf p = 0.84). (D) Serial killing by JR209 TCM and TEM of T2-A2Kb APCs loaded with 10μM gp100209-217(2M) at 1:3 T cell to APC ratio. Values represent the number of APCs killed as a percentage of total APCs, grouped by the number of APCs killed by each individual T cell. (E) Cytotoxic efficiency (the fraction of stable T cell/APC conjugates that resulted in APC lysis) of JR209 TCM and TEM when cultured with T2-A2Kb APCs loaded with 10μM gp100 at 1:1 T cell to APC ratio (n = 3, Two tailed t-test - p = 0.01, Difference in Proportions Test for each experiment - p < 0.03 for each).