The anti-tumor efficacy of adoptively transferred T cells requires their in vivo persistence and memory polarization. It is unknown if human chimeric antigen receptor (CAR)-expressing T cells can also undergo memory polarization. We examined the functional status of CAR CD8+ T cells, re-directed to Lewis Y antigen (LeY-T), throughout a period of ex vivo expansion. Immediately before culture CD8+ T cells comprised a mixture of phenotypes including naive (CD45RA+/CCR7+/CD27+/CD28+/perforin−), central memory (CM, CD45RA−/CCR7 lo/CD27+/CD28+/perforin lo), effector memory (EM, CD45RA−/CCR7−/CD27+/CD28+/perforin mod) and effector (Eff, CD45RA+/CCR7−/CD27−/CD28−/perforin hi) cells. After transduction and expansion culture of peripheral blood mononuclear cells from normal donors or multiple myeloma patients, CD8+ LeY-T cells polarized to EM-and CM-like phenotype. CD8+ LeY-T cells differed from starting CD8+ CM and EM T cells in that CD27, but not CD28, was downregulated. In addition, CD8+ LeY-T cells expressed high levels of perforin, similar to starting CD8+ Eff. CD8+ LeY-T cells also showed hallmarks of both memory and Eff function, underwent homeostatic proliferation in response to interleukin (IL)-15, and showed interferon (IFN)-γ production and cytotoxicity in response to Le-Y antigen on OVCAR-3 (human ovarian adenocarcinoma) cells. This study confirms CD8+ LeY-T cells have a CM-and EM-like phenotype and heterogeneous function consistent with potential to persist in vivo after adoptive transfer.