Interleukin-7 and interleukin-15 drive CD4+CD28null T lymphocyte expansion and function in patients with acute coronary syndrome

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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874 PATIENTS WITH ACUTE CORONARY SYNDROME SHOW OLIGOCLONAL T-CELL EXPANSION AND LESIONAL RECRUITMENT OF DIFFERENT T LYMPHOCYTE SUBTYPES

Atherosclerosis Supplements ◽

10.1016/s1567-5688(11)70875-3 ◽

2011 ◽

Vol 12 (1) ◽

pp. 183

Author(s):

S. Wanqler ◽

T.J. Dengler ◽

J. Haas ◽

B. Wildemann ◽

D. Bockler ◽

...

Keyword(s):

Acute Coronary Syndrome ◽

T Cell ◽

T Lymphocyte ◽

Cell Expansion ◽

Coronary Syndrome ◽

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

Immunology ◽

10.1111/j.1365-2567.2008.02843.x ◽

2008 ◽

Vol 125 (3) ◽

pp. 320-330 ◽

Cited By ~ 2

Author(s):

Nicolas Montcuquet ◽

Patricia Mercier-Letondal ◽

Sylvain Perruche ◽

Anne Duperrier ◽

Mélanie Couturier ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Regulatory T Cell ◽

Cell Expansion ◽

T Cell Expansion ◽

And Function

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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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B cell depletion impairs vaccination-induced CD8+ T cell responses in a type I interferon-dependent manner

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2021-220435 ◽

2021 ◽

pp. annrheumdis-2021-220435

Author(s):

Theresa Graalmann ◽

Katharina Borst ◽

Himanshu Manchanda ◽

Lea Vaas ◽

Matthias Bruhn ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

B Cells ◽

B Cell ◽

Cell Expansion ◽

De Novo ◽

T Cell Responses ◽

Type I ◽

Cell Responses ◽

T Cell Expansion

ObjectivesThe monoclonal anti-CD20 antibody rituximab is frequently applied in the treatment of lymphoma as well as autoimmune diseases and confers efficient depletion of recirculating B cells. Correspondingly, B cell-depleted patients barely mount de novo antibody responses during infections or vaccinations. Therefore, efficient immune responses of B cell-depleted patients largely depend on protective T cell responses.MethodsCD8+ T cell expansion was studied in rituximab-treated rheumatoid arthritis (RA) patients and B cell-deficient mice on vaccination/infection with different vaccines/pathogens.ResultsRituximab-treated RA patients vaccinated with Influvac showed reduced expansion of influenza-specific CD8+ T cells when compared with healthy controls. Moreover, B cell-deficient JHT mice infected with mouse-adapted Influenza or modified vaccinia virus Ankara showed less vigorous expansion of virus-specific CD8+ T cells than wild type mice. Of note, JHT mice do not have an intrinsic impairment of CD8+ T cell expansion, since infection with vaccinia virus induced similar T cell expansion in JHT and wild type mice. Direct type I interferon receptor signalling of B cells was necessary to induce several chemokines in B cells and to support T cell help by enhancing the expression of MHC-I.ConclusionsDepending on the stimulus, B cells can modulate CD8+ T cell responses. Thus, B cell depletion causes a deficiency of de novo antibody responses and affects the efficacy of cellular response including cytotoxic T cells. The choice of the appropriate vaccine to vaccinate B cell-depleted patients has to be re-evaluated in order to efficiently induce protective CD8+ T cell responses.

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Optimizing interleukin-2 concentration, seeding density and bead-to-cell ratio of T-cell expansion for adoptive immunotherapy

BMC Immunology ◽

10.1186/s12865-021-00435-7 ◽

2021 ◽

Vol 22 (1) ◽

Author(s):

Sasan Ghaffari ◽

Monireh Torabi-Rahvar ◽

Sajjad Aghayan ◽

Zahra Jabbarpour ◽

Kobra Moradzadeh ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Growth Kinetics ◽

Cell Expansion ◽

Mononuclear Cells ◽

Adoptive Cell Therapy ◽

Interleukin 2 ◽

Seeding Density ◽

Healthy Donors ◽

T Cell Expansion

Abstract Background The successful ex vivo expansion of T-cells in great numbers is the cornerstone of adoptive cell therapy. We aimed to achieve the most optimal T-cell expansion condition by comparing the expansion of T-cells at various seeding densities, IL-2 concentrations, and bead-to-cell ratios. we first expanded the peripheral blood mononuclear cells (PBMCs) of a healthy donor at a range of 20 to 500 IU/mL IL-2 concentrations, 125 × 103 to 1.5 × 106 cell/mL, and 1:10 to 10:1 B:C (Bead-to-cell) ratios and compared the results. We then expanded the PBMC of three healthy donors using the optimized conditions and examined the growth kinetics. On day 28, CD3, CD4, and CD8 expression of the cell populations were analyzed by flow cytometry. Results T-cells of the first donor showed greater expansion results in IL-2 concentrations higher than 50 IU/mL compared to 20 IU/mL (P = 0.02). A seeding density of 250 × 103 cell/mL was superior to higher or lower densities in expanding T-cells (P = 0.025). Also, we witnessed a direct correlation between the B:C ratio and T-cell expansion, in which, in 5:1 and 10:1 B:C ratios T-cell significantly expanded more than lower B:C ratios. The results of PBMC expansions of three healthy donors were similar in growth kinetics. In the optimized condition, 96–98% of the lymphocyte population expressed CD3. While the majority of these cells expressed CD8, the mean expression of CD4 in the donors was 19.3, 16.5, and 20.4%. Conclusions Our methodology demonstrates an optimized culture condition for the production of large quantities of polyclonal T-cells, which could be useful for future clinical and research studies.

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Polyclonal expansion of TCR Vbeta 21.3+ CD4+ and CD8+ T cells is a hallmark of Multisystem Inflammatory Syndrome in Children

Science Immunology ◽

10.1126/sciimmunol.abh1516 ◽

2021 ◽

Vol 6 (59) ◽

pp. eabh1516

Author(s):

Marion Moreews ◽

Kenz Le Gouge ◽

Samira Khaldi-Plassart ◽

Rémi Pescarmona ◽

Anne-Laure Mathieu ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cell Expansion ◽

Healthy Children ◽

Antigenic Peptides ◽

T Cell Repertoire ◽

Cell Repertoire ◽

Hla Dr ◽

T Cell Expansion ◽

Inflammatory Syndrome

Multiple Inflammatory Syndrome in Children (MIS-C) is a delayed and severe complication of SARS-CoV-2 infection that strikes previously healthy children. As MIS-C combines clinical features of Kawasaki disease and Toxic Shock Syndrome (TSS), we aimed to compare the immunological profile of pediatric patients with these different conditions. We analyzed blood cytokine expression, and the T cell repertoire and phenotype in 36 MIS-C cases, which were compared to 16 KD, 58 TSS, and 42 COVID-19 cases. We observed an increase of serum inflammatory cytokines (IL-6, IL-10, IL-18, TNF-α, IFNγ, CD25s, MCP1, IL-1RA) in MIS-C, TSS and KD, contrasting with low expression of HLA-DR in monocytes. We detected a specific expansion of activated T cells expressing the Vβ21.3 T cell receptor β chain variable region in both CD4 and CD8 subsets in 75% of MIS-C patients and not in any patient with TSS, KD, or acute COVID-19; this correlated with the cytokine storm detected. The T cell repertoire returned to baseline within weeks after MIS-C resolution. Vβ21.3+ T cells from MIS-C patients expressed high levels of HLA-DR, CD38 and CX3CR1 but had weak responses to SARS-CoV-2 peptides in vitro. Consistently, the T cell expansion was not associated with specific classical HLA alleles. Thus, our data suggested that MIS-C is characterized by a polyclonal Vβ21.3 T cell expansion not directed against SARS-CoV-2 antigenic peptides, which is not seen in KD, TSS and acute COVID-19.

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Mast cells and T-cell expansion

Blood ◽

10.1182/blood-2007-12-128330 ◽

2008 ◽

Vol 111 (5) ◽

pp. 2497-2498

Author(s):

Susumu Nakae ◽

Keisuke Oboki ◽

Hirohisa Saito

Keyword(s):

T Cells ◽

Dendritic Cells ◽

T Cell ◽

Mast Cells ◽

Cell Expansion ◽

Memory T Cells ◽

T Cell Expansion

IgE/antigen-FcϵRI crosslinking promotes antigen internalization and apoptosis in mouse mast cells. Dendritic cells uptake the apoptotic mast cells carrying internalized antigens, and thus can efficiently present the antigens to memory T cells.

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