TGFβ Programs Central Memory Differentiation in<i>Ex Vivo</i>–Stimulated Human T Cells

Dahmani, Amina; Janelle, Valérie; Carli, Cédric; Richaud, Manon; Lamarche, Caroline; Khalili, Myriam; Goupil, Mathieu; Bezverbnaya, Ksenia; Bramson, Jonathan L.; Delisle, Jean-Sébastien

doi:10.1158/2326-6066.cir-18-0691

Cited by 21 publications

(20 citation statements)

References 49 publications

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“…We generated B-cell maturation antigen (BCMA)-specific CAR T cells ( 40 ) that were activated and expanded for three weeks with anti-CD3/CD28 beads and subsequently transduced with p16 INK4a - targeting or non-targeting shRNAs ( Figures 7E, F ). While exposure to repetitive stimulations with BCMA-expressing human KMS-11 multiple myeloma cells ( 64 ) led to the expression of inhibitory receptors such as PD-1 and KLRG1 ( Supplementary Figure 5 ), p16 INK4a knockdown increased the fraction of Ki67 + cells and restricted the development of SA-β-Gal-expressing CAR T cells ( Figures 7G, H ). In contrast with data obtained with p38i, directly modulating p16 INK4a expression increased the proportion of cytokine-secreting T cells in this setting ( Figure 7I ).…”

Section: Resultsmentioning

confidence: 99%

p16INK4a Regulates Cellular Senescence in PD-1-Expressing Human T Cells

et al. 2021

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T-cell dysfunction arising upon repeated antigen exposure prevents effective immunity and immunotherapy. Using various clinically and physiologically relevant systems, we show that a prominent feature of PD-1-expressing exhausted T cells is the development of cellular senescence features both in vivo and ex vivo. This is associated with p16INK4a expression and an impaired cell cycle G1 to S-phase transition in repeatedly stimulated T cells. We show that these T cells accumulate DNA damage and activate the p38MAPK signaling pathway, which preferentially leads to p16INK4a upregulation. However, in highly dysfunctional T cells, p38MAPK inhibition does not restore functionality despite attenuating senescence features. In contrast, p16INK4a targeting can improve T-cell functionality in exhausted CAR T cells. Collectively, this work provides insights into the development of T-cell dysfunction and identifies T-cell senescence as a potential target in immunotherapy.

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Section: Resultsmentioning

confidence: 99%

p16INK4a Regulates Cellular Senescence in PD-1-Expressing Human T Cells

et al. 2021

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“… 30 , 31 TGF-β1 is a cytokine with pleiotropic effects, notably immunoregulatory and antiproliferative actions. It participates in T-cell development and differentiation into regulatory and central memory T cells among others, 32 , 33 and also participates in the healing process after tissue injury by acting in autocrine and paracrine signaling pathways. Without adequate negative feedback, its action can prove harmful owing to the accumulation of extracellular matrix.…”

Section: Discussionmentioning

confidence: 99%

Clinical Use of Complement, Inflammation, and Fibrosis Biomarkers in Autoimmune Glomerulonephritis

Khalili

Bonnefoy

Genest

et al. 2020

Kidney International Reports

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Introduction: Complement activation, inflammation, and fibrosis play central roles in the mechanisms of injury in autoimmune glomerulonephritis (GN) but they are seldom assessed in epidemiologic studies. The measurement of urinary biomarkers of these pathways of injury could parallel disease activity and add clinical value beyond proteinuria. Methods: We performed a prospective cohort study of 100 patients with focal and segmental glomerulosclerosis (FSGS), membranous nephropathy (MN), IgA nephropathy (IgAN), lupus nephritis (LN), antineutrophil cytoplasmic autoantibody-associated vasculitis (AAV), and membranoproliferative GN (MPGN) followed for 33 (18-54) months. Repeated urinary samples were collected throughout their follow-up to determine proteinuria, urinary sC5b-9, monocyte chemoattractant protein-1 (MCP-1), and transforming growth factor-beta 1 (TGF-b1), expressed as creatinine ratios. We identified 177 periods of active and inactive disease based on current remission definitions for each disease. Results: Urinary sC5b-9, MCP-1, and TGF-b1 were present in each disease. In periods leading to a remission, the reduction of urinary sC5b-9 was 91%, greater than for proteinuria with 76%. During inactive periods, those who did not experience a relapse maintained lower levels of biomarkers compared with those who relapsed. At that time, the increase in urinary sC5b-9 was significantly greater than the rise in proteinuria (8.5-fold increase compared with 3.2-fold) and urinary MCP-1 and TGF-b1. Using current remission definitions for each disease, thresholds for each biomarker were determined using receiver operating characteristic curves. Individuals who averaged levels below these cutoffs during their follow-up had better renal outcomes. Conclusion: In autoimmune glomerular diseases, urinary sC5b-9, MCP-1, and TGF-b1 are present and parallel disease activity and outcomes. Urinary sC5b-9 appears to be a more discerning marker of immunologic remissions and relapses.

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“…Flow cytometry-based cytotoxicity assay was performed using either Cell Trace Violet (CTV) or Cell Trace Yellow (CTY) (Invitrogen Life Technologies) labeled LMP2 426-434 pulsed as target cells as described before 14 . Briefly, target cells were prepared by stimulating autologous PBMCs with the T-cell mitogen phytohemagglutinin (PHA) (3 × 10 6 PBMCs/mL were incubated in T-cell media with 20 μg/mL PHA) for 3 days at 37 °C and 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

Combined PD-L1 and TIM-3 blockade improves the expansion of fit human CD8+ antigen-specific T cells for adoptive immunotherapy

Lak

Janelle

Djedid

et al. 2021

Preprint

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Background. The stimulation and expansion of antigen-specific T cells ex vivo enables the targeting of a multitude of cancer antigens. However, clinical scale T-cell expansion from rare precursors requires repeated stimulations ex vivo leading to T-cell terminal effector differentiation and exhaustion that adversely impact therapeutic potential. We leveraged immune checkpoint blockade relevant to antigen-specific CD8+ human T cells to improve the expansion and function of T cells targeting clinically relevant antigens. Methods. A clinically-compliant protocol relying on peptide-pulsed monocyte-derived dendritic cells and cytokines was used to expand antigen-specific CD8+ targeting the oncogenic Epstein-Barr virus (EBV) and the tumor associated antigen (TAA) Wilms Tumor 1 (WT1) protein. The effects of antibody-mediated blockade of immune checkpoints applied to the cultures (T-cell expansion, phenotypes and function) were assessed at various time points. Genomic studies including single cell RNA (scRNA) sequencing and T-cell receptor sequencing was performed on EBV-specific T cells to inform about the impact of immune checkpoint blockade on the clonal distribution and gene expression of the expanded T cells. Results. Several immune checkpoints were expressed early by ex vivo expanded antigen-specific CD8+ T cells, including PD-1 and TIM-3 with co-expression matching evidence of T-cell dysfunction as the cultures progressed. The introduction of anti-PD-L1 (expressed by the dendritic cells) and anti-TIM-3 antibodies in combination (but not individually) to the culture led to markedly improved antigen-specific T-cell expansion based on cell counts, fluorescent multimer staining and functional tests. This was not associated with evidence of T-cell dysfunction when compared to T cells expanded without immune checkpoint blockade. Genomic studies largely confirmed these results, showing that double blockade does not impart particular transcriptional programs or patterns on TCR repertoires. However, our data indicate that combined blockade may nonetheless alter gene expression in a minority of clonotypes and have donor-specific impacts. Conclusions. The manufacturing of antigen-specific CD8+ T cells can be improved in terms of yield and functionality using blockade of TIM-3 and the PD-L1/PD-1 axis in combination. Overcoming the deleterious effects of multiple antigenic stimulations through PD-L1/TIM-3 blockade is a readily applicable approach for several adoptive-immunotherapy strategies.

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TGFβ Programs Central Memory Differentiation inEx Vivo–Stimulated Human T Cells

Cited by 21 publications

References 49 publications

p16INK4a Regulates Cellular Senescence in PD-1-Expressing Human T Cells

p16INK4a Regulates Cellular Senescence in PD-1-Expressing Human T Cells

Clinical Use of Complement, Inflammation, and Fibrosis Biomarkers in Autoimmune Glomerulonephritis

Combined PD-L1 and TIM-3 blockade improves the expansion of fit human CD8+ antigen-specific T cells for adoptive immunotherapy

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