title: Phenotypic and functional attributes of lentivirus-modified CD19-specific human CD8+ central memory T cells manufactured at clinical scale.
abstract: A key determinant of the therapeutic potency of adoptive T-cell transfer is the extent to which infused cells can persist and expand in vivo. Ex vivo propagated virus-specific and chimeric antigen receptor (CAR)-redirected antitumor CD8 effector T cells derived from CD45RA(-) CD62L(+) central memory (TCM) precursors engraft long-term and reconstitute functional memory after adoptive transfer. Here, we describe a clinical scale, closed system, immunomagnetic selection method to isolate CD8(+) T(CM) from peripheral blood mononuclear cells (PBMC). This method uses the CliniMACS device to first deplete CD14(+), CD45RA(+), and CD4(+) cells from PBMC, and then to positively select CD62L(+) cells. The average purity and yield of CD8(+) CD45RA(-) CD62L TCM obtained in full-scale qualification runs were 70% and 0.4% (of input PBMC), respectively. These CD8(+) T(CM) are responsive to anti-CD3/CD28 bead stimulation, and can be efficiently transduced with CAR encoding lentiviral vectors, and undergo sustained expansion in interleukin (IL)-2/IL-15 over 3-6 weeks. The resulting CD8(+) T(CM)-derived effectors are polyclonal, retain expression of CD62L and CD28, exhibit CAR-redirected antitumor effector function, and are capable of huIL-15-dependent in vivo homeostatic engraftment after transfer to immunodeficient NOD/Scid IL-2RgCnull mice. Adoptive therapy using purified T(CM) cells is now the subject of a Food and Drug Administration-authorized clinical trial for the treatment of CD19(+) B-cell malignancies, and 3 clinical cell products expressing a CD19-specific CAR for IND #14645 have already been successfully generated from lymphoma patients using this manufacturing platform.
authors.affiliation: Departments of Cancer Immunotherapeutics & Tumor Immunology, and Hematology and Hematopoietic Cell Transplantation, Beckman Research Institute, City of Hope National Medical Center, Duarte, CA, USA.
authors.forename: Xiuli
authors.initials: X
authors.lastname: Wang
authors.forename: Araceli
authors.initials: A
authors.lastname: Naranjo
authors.forename: Christine E
authors.initials: CE
authors.lastname: Brown
authors.forename: Cherrilyn
authors.initials: C
authors.lastname: Bautista
authors.forename: Chinglam W
authors.initials: CW
authors.lastname: Wong
authors.forename: Wen-Chung
authors.initials: WC
authors.lastname: Chang
authors.forename: Brenda
authors.initials: B
authors.lastname: Aguilar
authors.forename: Julie R
authors.initials: JR
authors.lastname: Ostberg
authors.forename: Stanley R
authors.initials: SR
authors.lastname: Riddell
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Forman
authors.forename: Michael C
authors.initials: MC
authors.lastname: Jensen
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D018941:Antigens, CD19 / Q000378:metabolism*; D018106:CD28 Antigens / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D045744:Cell Line, Tumor; D015925:Cryopreservation; D016207:Cytokines / Q000096:biosynthesis; D003602:Cytotoxicity, Immunologic; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007156:Immunologic Memory; D018189:Immunomagnetic Separation / Q000379:methods; D016130:Immunophenotyping; D019041:L-Selectin / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D008213:Lymphocyte Activation / Q000276:immunology; D008223:Lymphoma / Q000276:immunology / Q000628:therapy; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D010641:Phenotype; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D014161:Transduction, Genetic; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7 / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
chemical_list: D018941:Antigens, CD19; D018106:CD28 Antigens; D017252:CD3 Complex; D016207:Cytokines; D011948:Receptors, Antigen, T-Cell; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7; D019041:L-Selectin