Regulatory T-cell expansion and function do not account for the impaired alloreactivity ofex vivo-expanded T cells

Abstract Aims Inflammation has important roles in atherosclerosis. CD4+CD28null (CD28null) T cells are a specialized T lymphocyte subset that produce inflammatory cytokines and cytotoxic molecules. CD28null T cells expand preferentially in patients with acute coronary syndrome (ACS) rather than stable angina and are barely detectable in healthy subjects. Importantly, ACS patients with CD28null T-cell expansion have increased risk for recurrent acute coronary events and poor prognosis, compared to ACS patients in whom this cell subset does not expand. The mechanisms regulating CD28null T-cell expansion in ACS remain elusive. We therefore investigated the role of cytokines in CD28null T-cell expansion in ACS. Methods and results High-purity sorted CD4+ T cells from ACS patients were treated with a panel of cytokines (TNF-α, IL-1β, IL-6, IL-7, and IL-15), and effects on the number, phenotype, and function of CD28null T cells were analysed and compared to the control counterpart CD28+ T-cell subset. IL-7- and IL-15-induced expansion of CD28null T cells from ACS patients, while inflammatory cytokines TNF-α, IL-1β, and IL-6 did not. The mechanisms underlying CD28null T-cell expansion by IL-7/IL-15 were preferential activation and proliferation of CD28null T cells compared to control CD28+ T cells. Additionally, IL-7/IL-15 markedly augmented CD28null T-cell cytotoxic function and interferon-γ production. Further mechanistic analyses revealed differences in baseline expression of component chains of IL-7/IL-15 receptors (CD127 and CD122) and increased baseline STAT5 phosphorylation in CD28null T cells from ACS patients compared to the control CD28+ T-cell subset. Notably, we demonstrate that CD28null T-cell expansion was significantly inhibited by Tofacitinib, a selective JAK1/JAK3 inhibitor that blocks IL-7/IL-15 signalling. Conclusion Our novel data show that IL-7 and IL-15 drive the expansion and function of CD28null T cells from ACS patients suggesting that IL-7/IL-15 blockade may prevent expansion of these cells and improve patient outcomes.

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Effector T Cells Boost Regulatory T Cell Expansion by IL-2, TNF, OX40, and Plasmacytoid Dendritic Cells Depending on the Immune Context

The Journal of Immunology ◽

10.4049/jimmunol.1400504 ◽

2014 ◽

Vol 194 (3) ◽

pp. 999-1010 ◽

Cited By ~ 29

Author(s):

Audrey Baeyens ◽

David Saadoun ◽

Fabienne Billiard ◽

Angéline Rouers ◽

Sylvie Grégoire ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

T Cell ◽

Regulatory T Cell ◽

Cell Expansion ◽

Plasmacytoid Dendritic Cells ◽

Effector T Cells ◽

T Cell Expansion

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

10.1101/2021.08.30.21262835 ◽

2021 ◽

Author(s):

Shirin Lak ◽

Valérie Janelle ◽

Anissa Djedid ◽

Gabrielle Boudreau ◽

Ann Brasey ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Immune Checkpoint ◽

Cell Expansion ◽

Ex Vivo ◽

Immune Checkpoint Blockade ◽

Checkpoint Blockade ◽

Immune Checkpoints ◽

T Cell Expansion ◽

And Function

AbstractBackgroundThe 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.MethodsA 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 were 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.ResultsSeveral 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. Genomics studies largely confirmed these results, showing that double blockade does not impart specific 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.ConclusionsThe 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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Clinically-relevant T cell expansion protocols activate distinct cellular metabolic programs and phenotypes

10.1101/2021.08.24.457536 ◽

2021 ◽

Author(s):

Sarah MacPherson ◽

Sarah Keyes ◽

Marisa K Kilgour ◽

Julian Smazynski ◽

Jessica Sudderth ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cell Expansion ◽

Cell Function ◽

Ex Vivo ◽

Ex Vivo Expansion ◽

Metabolic Phenotypes ◽

T Cell Function ◽

T Cell Expansion ◽

And Function

Ex vivo expansion conditions used to generate T cells for immunotherapy are thought to adopt metabolic phenotypes that impede therapeutic efficacy in vivo. The comparison of five different culture media used for clinical T cell expansion revealed unique optima based on different output variables including proliferation, differentiation, function, activation and mitochondrial phenotypes. T cells adapted their metabolism to match their media expansion condition as shown by glucose and glutamine uptake, and patterns of glucose isotope labeling. However, adoption of these metabolic phenotypes was uncoupled to T cell function. Furthermore, T cell products cultured in ascites from ovarian cancer patients displayed suppressed mitochondrial activity and function irrespective of the ex vivo expansion media. In one case, culturing in ascites resulted in increased glucose uptake which was insufficient to rescue T cell function. Thus, ex vivo T cell expansion conditions have profound impacts on metabolism and function.

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Downregulation of IL-17-producing T cells is associated with regulatory T cell expansion and disease progression in chronic lymphocytic leukemia

Tumor Biology ◽

10.1007/s13277-012-0628-4 ◽

2012 ◽

Vol 34 (2) ◽

pp. 929-940 ◽

Cited By ~ 42

Author(s):

Farhad Jadidi-Niaragh ◽

Ghasem Ghalamfarsa ◽

Ali Memarian ◽

Hossein Asgarian-Omran ◽

Seyed Mohsen Razavi ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Chronic Lymphocytic Leukemia ◽

Disease Progression ◽

Regulatory T Cell ◽

Cell Expansion ◽

Lymphocytic Leukemia ◽

T Cell Expansion

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Characterization of regulatory T cell expansion for manufacturing cellular immunotherapies

Biomaterials Science ◽

10.1039/d0bm00622j ◽

2020 ◽

Vol 8 (15) ◽

pp. 4186-4198 ◽

Cited By ~ 2

Author(s):

David A. McBride ◽

Matthew D. Kerr ◽

Shinya L. Wai ◽

Yvonne Y. Yee ◽

Dora A. Ogbonna ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Regulatory T Cells ◽

Regulatory T Cell ◽

Cell Expansion ◽

Beta Cyclodextrin ◽

T Cell Expansion

Rapamycin encapsulated in mono-(6-amino-6-deoxy)-beta cyclodextrin efficiently expands regulatory T cells for cell-based immunotherapy.

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Downregulation of IL-17-producing T cells is associated with regulatory T cell expansion and disease progression in chronic lymphocytic leukemia

Frontiers in Immunology ◽

10.3389/conf.fimmu.2013.02.00084 ◽

2013 ◽

Vol 4 ◽

Author(s):

Jadidi-Niaragh Farhad ◽

Ghalamfarsa Ghasem ◽

Memarian Ali ◽

Asgarian-Omran Hossein ◽

Razavi Seyed Mohsen ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Chronic Lymphocytic Leukemia ◽

Disease Progression ◽

Regulatory T Cell ◽

Cell Expansion ◽

Lymphocytic Leukemia ◽

T Cell Expansion

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Faculty Opinions recommendation of Foxp3(+) regulatory T cell expansion required for sustaining pregnancy compromises host defense against prenatal bacterial pathogens.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.13085956.14411054 ◽

2011 ◽

Author(s):

Victor Nizet

Keyword(s):

T Cell ◽

Host Defense ◽

Regulatory T Cell ◽

Cell Expansion ◽

Bacterial Pathogens ◽

T Cell Expansion

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764 Expansion with IL-15 increases cytotoxicity of Vγ9Vδ2 T cells and is associated with higher levels of cytotoxic molecules and T-bet

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-sitc2020.0764 ◽

2020 ◽

Vol 8 (Suppl 3) ◽

pp. A812-A812

Author(s):

Pia Aehnlich ◽

Per Thor Straten ◽

Ana Micaela Carnaz Simoes ◽

Signe Skadborg ◽

Gitte Olofsson

Keyword(s):

T Cells ◽

T Cell ◽

Cell Expansion ◽

Ex Vivo ◽

Adoptive Cell Therapy ◽

Hematological Cancers ◽

Cytotoxic Molecules ◽

Vγ9vδ2 T Cells ◽

T Cell Expansion

BackgroundAdoptive cell therapy (ACT) is an approved treatment option for certain hematological cancers and has also shown success for some solid cancers. Still, benefit and eligibility do not extend to all patients. ACT with Vγ9Vδ2 T cells is a promising approach to overcome this hurdle.MethodsIn this study, we explored the effect of different cytokine conditions on the expansion of Vγ9Vδ2 T cells in vitro.ResultsWe could show that Vγ9Vδ2 T cell expansion is feasible with two different cytokine conditions: (a) 1000U/ml interleukin (IL)-2 and (b) 100U/ml IL-2+100U/ml IL-15. We did not observe differences in expansion rate or Vγ9Vδ2 T cell purity between the conditions; however, IL-2/IL-15-expanded Vγ9Vδ2 T cells displayed enhanced cytotoxicity against tumor cells, also in hypoxia. While this increase in killing capacity was not reflected in phenotype, we demonstrated that IL-2/IL-15-expanded Vγ9Vδ2 T cells harbor increased amounts of perforin, granzyme B and granulysin in a resting state and release more upon activation. IL-2/IL-15-expanded Vγ9Vδ2 T cells also showed higher levels of transcription factor T-bet, which could indicate that T-bet and cytotoxic molecule levels confer the increased cytotoxicity.ConclusionsThese results advocate the inclusion of IL-15 into ex vivo Vγ9Vδ2 T cell expansion protocols in future clinical studies.

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