Phenotype of CAR⁺ T cell.

<p>(A) Expression of CD19RCD28 CAR on T cells day after electroporation (culture day 1) and after 28 days of co-culture on aAPC clone #4 along with lack of CD19⁺ aAPC. (B) CAR expression by western blot analysis using CD3-ζ specific antibody. Whole cell lysates were run on SDS-PAGE under reducing conditions. Molecular weight marker (M), Parental Jurkat cells (Lane 1), CD19RCD28⁺ Jurkat cells (Lane 2), CAR^neg control T cells (Lane 3) and CD19RCD28⁺ T cells (Lane 4). (C) Percent expression of CD3⁺, CD4⁺CAR⁺ and CD8⁺CAR⁺ T cells with in a lymphocyte gate in cultures over time. Each symbol represents a separate experiment; the solid lines are mean of the three validation experiments. (D) Immunophenotype of memory/naïve, adhesion, activation, cytolytic and exhaustion markers on CAR⁺ T cells at the end (d28) of co-culture.