title: Reprogramming CD19-specific T cells with IL-21 signaling can improve adoptive immunotherapy of B-lineage malignancies.
abstract: Improving the therapeutic efficacy of T cells expressing a chimeric antigen receptor (CAR) represents an important goal in efforts to control B-cell malignancies. Recently an intrinsic strategy has been developed to modify the CAR itself to improve T-cell signaling. Here we report a second extrinsic approach based on altering the culture milieu to numerically expand CAR(+) T cells with a desired phenotype, for the addition of interleukin (IL)-21 to tissue culture improves CAR-dependent T-cell effector functions. We used electrotransfer of Sleeping Beauty system to introduce a CAR transposon and selectively propagate CAR(+) T cells on CD19(+) artificial antigen-presenting cells (aAPC). When IL-21 was present, there was preferential numeric expansion of CD19-specific T cells which lysed and produced IFN-γ in response to CD19. Populations of these numerically expanded CAR(+) T cells displayed an early memory surface phenotype characterized as CD62L(+)CD28(+) and a transcriptional profile of naïve T cells. In contrast, T cells propagated with only exogenous IL-2 tended to result in an overgrowth of CD19-specific CD4(+) T cells. Furthermore, adoptive transfer of CAR(+) T cells cultured with IL-21 exhibited improved control of CD19(+) B-cell malignancy in mice. To provide coordinated signaling to propagate CAR(+) T cells, we developed a novel mutein of IL-21 bound to the cell surface of aAPC that replaced the need for soluble IL-21. Our findings show that IL-21 can provide an extrinsic reprogramming signal to generate desired CAR(+) T cells for effective immunotherapy.
authors.affiliation: Division of Pediatrics, Children's Cancer Hospital, Houston, Texas, USA.
authors.forename: Harjeet
authors.initials: H
authors.lastname: Singh
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Figliola
authors.forename: Margaret J
authors.initials: MJ
authors.lastname: Dawson
authors.forename: Helen
authors.initials: H
authors.lastname: Huls
authors.forename: Simon
authors.initials: S
authors.lastname: Olivares
authors.forename: Kirsten
authors.initials: K
authors.lastname: Switzer
authors.forename: Tiejuan
authors.initials: T
authors.lastname: Mi
authors.forename: Sourindra
authors.initials: S
authors.lastname: Maiti
authors.forename: Partow
authors.initials: P
authors.lastname: Kebriaei
authors.forename: Dean A
authors.initials: DA
authors.lastname: Lee
authors.forename: Richard E
authors.initials: RE
authors.lastname: Champlin
authors.forename: Laurence J N
authors.initials: LJ
authors.lastname: Cooper
mesh_terms: D000818:Animals; D018941:Antigens, CD19 / Q000096:biosynthesis* / Q000378:metabolism; D001402:B-Lymphocytes / Q000166:cytology*; D018106:CD28 Antigens / Q000378:metabolism; D019070:Cell Lineage; D018920:Coculture Techniques; D019337:Hematologic Neoplasms / Q000378:metabolism*; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007371:Interferon-gamma / Q000378:metabolism; D018664:Interleukin-12 / Q000378:metabolism*; D007378:Interleukins / Q000378:metabolism; D020014:K562 Cells; D019041:L-Selectin / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D050796:STAT3 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D000098546:Interleukin-21
chemical_list: D018941:Antigens, CD19; D018106:CD28 Antigens; D007378:Interleukins; D050796:STAT3 Transcription Factor; D019041:L-Selectin; D018664:Interleukin-12; D007371:Interferon-gamma; D000098546:Interleukin-21