In vitro generation of mature, naive antigen-specific CD8+ T cells with a single T-cell receptor by agonist selection

Abstract

Peripheral blood T cells transduced with a tumor-specific T-cell receptor (TCR) face problems of auto-reactivity and lack of efficacy caused by cross-pairing of exogenous and endogenous TCR chains, as well as short term in vivo survival due to activation and growth factor-induced differentiation. We here studied an alternative strategy for the efficient generation of naive CD8(+) T cells with a single TCR. TCR-transduced human postnatal thymus-derived and adult mobilized blood-derived hematopoietic progenitor cells (HPCs) were differentiated to CD4(+)CD8(+) double-positive T cells using OP9-Delta-like 1 (OP9-DL1) cultures. Addition of the agonist peptide induced double positive cells to cross-present the peptide, leading, in the absence of co-stimulation, to cell cycle arrest and differentiation into mature CD8(+) T cells. Comprehensive phenotypic, molecular and functional analysis revealed the generation of naive and resting CD8(+) T cells through a process similar to thymic positive selection. These mature T cells show a near complete inhibition of endogenous TCRA and TCRB rearrangements and express high levels of the introduced multimer-reactive TCR. Upon activation, specific cytokine production and efficient killing of tumor cells were induced. Using this strategy, large numbers of high-avidity tumor-specific naive T cells can be generated from readily available HPCs without TCR chain cross-pairing.

Ginsenoside compound K suppresses the abnormal activation of T lymphocytes in mice with collagen-induced arthritis

“…Simultaneous recognition of the cognate MHC-peptide complex by the TCR (signal 1) and CD80 or CD86 by CD28 (signal 2) results in T cell activation, proliferation, and differentiation [10,11] . In addition to cytokine production, PD-1 and CTLA-4 are inducibly expressed on T cells following a TCR signal, and subsequent binding of the TCR to one of these co-inhibitors results in cell cycle arrest and termination of T cell activation [12][13][14] .…”

Generation of Tumor Antigen-Specific iPSC-Derived Thymic Emigrants Using a 3D Thymic Culture System

“…Current in vitro iPSC differentiation methods employ strong TCR signaling triggered by either anti-CD3 or anti-TCR antibodies (Nishimura et al, 2013; Vizcardo et al, 2013) or by agonist peptides (Snauwaert et al, 2014). However, T cells induced by TCR stimulation or high-affinity peptides are generally incompetent functionally or are driven into unconventional T cell lineages, including regulatory T cells, natural killer (NK) T cells, and CD8αα + T cells (Takada et al, 2017; Yamagata et al, 2004).…”

Antigen receptor-redirected T cells derived from hematopoietic precursor cells lack expression of the endogenous TCR/CD3 receptor and exhibit specific antitumor capacities

“…In addition, we have shown that in cultures initiated with HPCs transduced to express a TCRαβ, mature CD27 + CD1a − T cells are virtually absent, but addition of the agonist peptide in the presence of the restricting HLA antigen induces maturation 27 . Here, antigen-dependent maturation is unlikely as CEA expression analysis on these cultures with qPCR was consistently negative (data not shown).…”

“…We previously reported that mono-specific TCR-transgenic cells lacking endogenous rearrangements can be generated from cord blood hematopoietic progenitor cells (HPCs) 27 . This method is based on allelic exclusion, a phenomenon that occurs physiologically in the thymus and that ensures that each T cell successfully rearranges only a single TCRβ locus.…”