title: Bispecific T-cells expressing polyclonal repertoire of endogenous γδ T-cell receptors and introduced CD19-specific chimeric antigen receptor.
abstract: Even though other γδ T-cell subsets exhibit antitumor activity, adoptive transfer of γδ Tcells is currently limited to one subset (expressing Vγ9Vδ2 T-cell receptor (TCR)) due to dependence on aminobisphosphonates as the only clinically appealing reagent for propagating γδ T cells. Therefore, we developed an approach to propagate polyclonal γδ T cells and rendered them bispecific through expression of a CD19-specific chimeric antigen receptor (CAR). Peripheral blood mononuclear cells (PBMC) were electroporated with Sleeping Beauty (SB) transposon and transposase to enforce expression of CAR in multiple γδ T-cell subsets. CAR(+)γδ T cells were expanded on CD19(+) artificial antigen-presenting cells (aAPC), which resulted in >10(9) CAR(+)γδ T cells from <10(6) total cells. Digital multiplex assay detected TCR mRNA coding for Vδ1, Vδ2, and Vδ3 with Vγ2, Vγ7, Vγ8, Vγ9, and Vγ10 alleles. Polyclonal CAR(+)γδ T cells were functional when TCRγδ and CAR were stimulated and displayed enhanced killing of CD19(+) tumor cell lines compared with CAR(neg)γδ T cells. CD19(+) leukemia xenografts in mice were reduced with CAR(+)γδ T cells compared with control mice. Since CAR, SB, and aAPC have been adapted for human application, clinical trials can now focus on the therapeutic potential of polyclonal γδ T cells.
authors.affiliation: The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: Drew C
authors.initials: DC
authors.lastname: Deniger
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: Lenka
authors.initials: L
authors.lastname: Hurton
authors.forename: Harjeet
authors.initials: H
authors.lastname: Singh
authors.forename: Helen
authors.initials: H
authors.lastname: Huls
authors.forename: Simon
authors.initials: S
authors.lastname: Olivares
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; D000938:Antigen-Presenting Cells / Q000276:immunology / Q000378:metabolism; D018941:Antigens, CD19 / Q000276:immunology* / Q000378:metabolism; D045744:Cell Line, Tumor; D018274:Electroporation; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007938:Leukemia / Q000628:therapy; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D018345:Mice, Knockout; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*; D019895:Transposases / Q000235:genetics / Q000378:metabolism
chemical_list: D018941:Antigens, CD19; D016692:Receptors, Antigen, T-Cell, gamma-delta; D019895:Transposases; C494471:sleeping beauty transposase, human