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Bispecific T-cells expressing polyclonal repertoire of endogenous ?? T-cell receptors and introduced CD19-specific chimeric antigen receptor.

ABSTRACT: Even though other ?? T-cell subsets exhibit antitumor activity, adoptive transfer of ?? Tcells is currently limited to one subset (expressing V?9V?2 T-cell receptor (TCR)) due to dependence on aminobisphosphonates as the only clinically appealing reagent for propagating ?? T cells. Therefore, we developed an approach to propagate polyclonal ?? 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 ?? T-cell subsets. CAR(+)?? T cells were expanded on CD19(+) artificial antigen-presenting cells (aAPC), which resulted in >10(9) CAR(+)?? T cells from <10(6) total cells. Digital multiplex assay detected TCR mRNA coding for V?1, V?2, and V?3 with V?2, V?7, V?8, V?9, and V?10 alleles. Polyclonal CAR(+)?? T cells were functional when TCR?? and CAR were stimulated and displayed enhanced killing of CD19(+) tumor cell lines compared with CAR(neg)?? T cells. CD19(+) leukemia xenografts in mice were reduced with CAR(+)?? 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 ?? T cells.

SUBMITTER: Deniger DC

PROVIDER: S-EPMC3589159 | biostudies-literature | 2013 Mar

REPOSITORIES: biostudies-literature

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Bispecific T-cells expressing polyclonal repertoire of endogenous γδ T-cell receptors and introduced CD19-specific chimeric antigen receptor.

Deniger Drew C DC Switzer Kirsten K Mi Tiejuan T Maiti Sourindra S Hurton Lenka L Singh Harjeet H Huls Helen H Olivares Simon S Lee Dean A DA Champlin Richard E RE Cooper Laurence J N LJ

Molecular therapy : the journal of the American Society of Gene Therapy 20130108 3

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 ...[more]

PMID: 23295945

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Project description:BACKGROUND:Chimeric antigen receptor (CAR) modified T cells targeting CD19 have shown activity in case series of patients with acute and chronic lymphocytic leukaemia and B-cell lymphomas, but feasibility, toxicity, and response rates of consecutively enrolled patients treated with a consistent regimen and assessed on an intention-to-treat basis have not been reported. We aimed to define feasibility, toxicity, maximum tolerated dose, response rate, and biological correlates of response in children and young adults with refractory B-cell malignancies treated with CD19-CAR T cells. METHODS:This phase 1, dose-escalation trial consecutively enrolled children and young adults (aged 1-30 years) with relapsed or refractory acute lymphoblastic leukaemia or non-Hodgkin lymphoma. Autologous T cells were engineered via an 11-day manufacturing process to express a CD19-CAR incorporating an anti-CD19 single-chain variable fragment plus TCR zeta and CD28 signalling domains. All patients received fludarabine and cyclophosphamide before a single infusion of CD19-CAR T cells. Using a standard 3?+?3 design to establish the maximum tolerated dose, patients received either 1?×?10(6) CAR-transduced T cells per kg (dose 1), 3?×?10(6) CAR-transduced T cells per kg (dose 2), or the entire CAR T-cell product if sufficient numbers of cells to meet the assigned dose were not generated. After the dose-escalation phase, an expansion cohort was treated at the maximum tolerated dose. The trial is registered with ClinicalTrials.gov, number NCT01593696. FINDINGS:Between July 2, 2012, and June 20, 2014, 21 patients (including eight who had previously undergone allogeneic haematopoietic stem-cell transplantation) were enrolled and infused with CD19-CAR T cells. 19 received the prescribed dose of CD19-CAR T cells, whereas the assigned dose concentration could not be generated for two patients (90% feasible). All patients enrolled were assessed for response. The maximum tolerated dose was defined as 1?×?10(6) CD19-CAR T cells per kg. All toxicities were fully reversible, with the most severe being grade 4 cytokine release syndrome that occurred in three (14%) of 21 patients (95% CI 3·0-36·3). The most common non-haematological grade 3 adverse events were fever (nine [43%] of 21 patients), hypokalaemia (nine [43%] of 21 patients), fever and neutropenia (eight [38%] of 21 patients), and cytokine release syndrome (three [14%) of 21 patients). INTERPRETATION:CD19-CAR T cell therapy is feasible, safe, and mediates potent anti-leukaemic activity in children and young adults with chemotherapy-resistant B-precursor acute lymphoblastic leukaemia. All toxicities were reversible and prolonged B-cell aplasia did not occur. FUNDING:National Institutes of Health Intramural funds and St Baldrick's Foundation.

Dataset Information

Bispecific T-cells expressing polyclonal repertoire of endogenous ?? T-cell receptors and introduced CD19-specific chimeric antigen receptor.

Publications

Bispecific T-cells expressing polyclonal repertoire of endogenous γδ T-cell receptors and introduced CD19-specific chimeric antigen receptor.

Similar Datasets

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