title: Ex vivo culture of chimeric antigen receptor T cells generates functional CD8+ T cells with effector and central memory-like phenotype.
abstract: 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.
authors.affiliation: Hematology and Immunology Translational Research Lab, Peter MacCallum Cancer Center, Melbourne, Australia. paul.neeson@petermac.org
authors.forename: P
authors.initials: P
authors.lastname: Neeson
authors.forename: A
authors.initials: A
authors.lastname: Shin
authors.forename: K M
authors.initials: KM
authors.lastname: Tainton
authors.forename: P
authors.initials: P
authors.lastname: Guru
authors.forename: H M
authors.initials: HM
authors.lastname: Prince
authors.forename: S J
authors.initials: SJ
authors.lastname: Harrison
authors.forename: S
authors.initials: S
authors.lastname: Peinert
authors.forename: M J
authors.initials: MJ
authors.lastname: Smyth
authors.forename: J A
authors.initials: JA
authors.lastname: Trapani
authors.forename: M H
authors.initials: MH
authors.lastname: Kershaw
authors.forename: P K
authors.initials: PK
authors.lastname: Darcy
authors.forename: D S
authors.initials: DS
authors.lastname: Ritchie
mesh_terms: D018106:CD28 Antigens / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D049109:Cell Proliferation; D003602:Cytotoxicity, Immunologic; D006801:Humans; D007156:Immunologic Memory; D007371:Interferon-gamma / Q000378:metabolism; D017493:Leukocyte Common Antigens / Q000276:immunology; D010641:Phenotype; D011946:Receptors, Antigen / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7 / Q000276:immunology
chemical_list: D018106:CD28 Antigens; D011946:Receptors, Antigen; D011993:Recombinant Fusion Proteins; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7; D007371:Interferon-gamma; D017493:Leukocyte Common Antigens