Intermittent calorie restriction alters T cell subsets and metabolic markers in people with multiple sclerosis

Abstract Background: Intermittent fasting or calorie restriction (CR) diets provide anti-inflammatory and neuroprotective advantages in models of multiple sclerosis (MS); data in humans are sparse. Methods: We conducted a randomized-controlled feeding study of different CR diets in 36 people with MS over 8 weeks. Patients were randomized to receive either: a daily CR diet (22% reduction in calories, 7 days/week), an intermittent CR diet (75% reduction, 2 days/week; 100%, 5 days/week), or a weight-stable diet (100%, 7 days/week). Untargeted metabolomics was performed on plasma samples at weeks 0, 4 and 8 at Metabolon Inc (Durham, NC). Flow cytometry of cryopreserved peripheral blood mononuclear cells at weeks 0 and 8 were used to identify CD4+ and CD8+ T cell subsets including effector memory, central memory, and naïve cells. Results: 31 (86%) completed the trial. Over time, individuals randomized to intermittent CR had significant reductions in CD4+CM -4.87%; 95%CI: -8.59%, -1.15%; p=0.01), CD4+EM (-3.82%; 95%CI: -7.44, -0.21; p=0.04), and CD8+EM (-6.96%; 95%CI: -11.96, -1.97; p=0.006) with proportional increases in naïve subsets (CD4+Naïve: 5.81%; 95%CI: -0.01, 11.63%; p=0.05; CD8+Naïve: 10.11%; 95%CI: 3.30, 16.92%; p=0.006). No changes were observed for daily CR or weight-stable diets. Larger within-person changes in lysophospholipid and lysoplasmalogen metabolites in intermittent CR were associated with larger reductions in memory T cell subsets and larger increases in naïve T cell subsets. Conclusions: In people with MS, an intermittent CR diet was associated with reduction in memory T cell subsets. The observed changes may be mediated by changes in specific classes of lipid metabolites. Trial Registration: This study is registered on Clinicaltrials.gov with identifier NCT02647502. Funding: National MS Society, NIH, Johns Hopkins Catalyst Award

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CD4+T cells in ageing-associated interstitial lung abnormalities show evidence of pro-inflammatory phenotypic and functional profile

Thorax ◽

10.1136/thoraxjnl-2020-215520 ◽

2020 ◽

pp. thoraxjnl-2020-215520

Author(s):

Carlos Machahua ◽

Ivette Buendia-Roldan ◽

Ranferi Ocaña-Guzman ◽

María Molina-Molina ◽

Annie Pardo ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cd4 T Cells ◽

Mononuclear Cells ◽

Killer Cell ◽

T Cell Subsets ◽

Effector Memory ◽

Peripheral Blood Mononuclear ◽

Lung Abnormalities ◽

Immune Deficiencies

BackgroundInterstitial lung abnormalities (ILA) occur in around 10% of subjects over 60 years, and are associated with a higher rate of all-cause mortality. The pathogenic mechanisms are unclear, and the putative contribution of alterations in the immune response has not been explored. Normal ageing is associated with immune deficiencies, including Naïve T-cell decrease and greater expression of the proliferative-limiting, co-inhibitory receptor killer-cell lectin-like receptor G1 (KLRG1).ObjectiveTo evaluate the frequency and activation state of different T-cell subpopulations in ILA subjects.MethodsPeripheral blood mononuclear cells were obtained from 15 individuals with ILA, 21 age-matched controls and 28 healthy young subjects. T-cells phenotype was characterised by flow cytometry, and proliferation and activation by stimulation with anti-CD3/anti-CD28 or phorbol myristate acetate/ionomycin; KLRG1 isoforms were evaluated by western blot and cytokines were quantified by ELISA and Multiplex.ResultsA significant increase of Naïve CD4+T cells together with a decrease of central and effector memory CD4+T cells was observed in ILA compared with age-matched controls. CD4+T cells from ILA subjects exhibited greater basal proliferation, which raised after anti-CD3/anti-CD28 stimulation. Additionally, a significant increase in the levels of interleukin-6 and interferon gamma was observed in isolated CD4+T cells and plasma of ILA subjects. They also displayed fewer KLRG1+/CD4+T cells with an increase of circulating E-cadherin, the ligand of KLRG1+. No changes were observed with CD8+T cell subsets.ConclusionCD4+T cells from ILA subjects are highly proliferative and show an excessive functional activity, likely related to the loss of KLRG1 expression, which may contribute to an inflammatory state and the development of ILA.

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Individual Immune-Modulatory Capabilities of MSC-Derived Extracellular Vesicle (EV) Preparations and Recipient-Dependent Responsiveness

International Journal of Molecular Sciences ◽

10.3390/ijms20071642 ◽

2019 ◽

Vol 20 (7) ◽

pp. 1642 ◽

Cited By ~ 13

Author(s):

Lambros Kordelas ◽

Esther Schwich ◽

Robin Dittrich ◽

Peter Horn ◽

Dietrich Beelen ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Mononuclear Cells ◽

Therapeutic Option ◽

T Cell Response ◽

Cytokine Profile ◽

T Cell Subsets ◽

Effector Memory ◽

Peripheral Blood Mononuclear ◽

Vitro Assay

Treatment with extracellular vesicles (EVs) derived from mesenchymal stem/stromal cells (MSCs) have been suggested as novel therapeutic option in acute inflammation-associated disorders due to their immune-modulatory capacities. As we have previously observed differences in the cytokine profile of independent MSC-EV preparations, functional differences of MSC-EV preparations have to be considered. To evaluate the immune-modulatory capabilities of specific MSC-EV preparations, reliable assays are required to characterize the functionality of MSC-EV preparations prior to administration to a patient. To this end, we established an in vitro assay evaluating the immune-modulatory capacities of MSC-EV preparations. Here, we compared the efficacy of four independent MSC-EV preparations to modulate the induction of T cell differentiation and cytokine production after phorbol 12-myristate 13-acetate (PMA)/Ionomycin stimulation of peripheral blood mononuclear cells (PBMC) derived from six healthy donors. Flow cytometric analyses revealed that the four MSC-EV preparations differentially modulate the expression of surface markers, such as CD45RA, on CD4+ and CD8+ T cells, resulting in shifts in the frequencies of effector and effector memory T cells. Moreover, cytokine profile in T cell subsets was affected in a MSC-EV-specific manner exclusively in CD8+ naïve T cells. Strikingly, hierarchical clustering revealed that the T cell response towards the MSC-EV preparations largely varied among the different PBMC donors. Thus, besides defining functional activity of MSC-EV preparations, it will be crucial to test whether patients intended for treatment with MSC-EV preparations are in principal competent to respond to the envisioned MSC-EV therapy.

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Both Naïve and Memory T-Cell Subsets Participate in Alloresponses to HLA-Mismatched Targets – Implications for Adoptive Transfer of Viral Antigen-Specific T Cells.

Blood ◽

10.1182/blood.v114.22.2439.2439 ◽

2009 ◽

Vol 114 (22) ◽

pp. 2439-2439

Author(s):

J. Joseph Melenhorst ◽

Phillip Scheinberg ◽

Ann Williams ◽

Keyvan Keyvanfar ◽

Melody Smith ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Viral Antigen ◽

Mononuclear Cells ◽

Memory T Cells ◽

T Cell Subsets ◽

Dna Virus ◽

Activated T Cells ◽

Peripheral Blood Mononuclear ◽

Memory T Cell

Abstract Abstract 2439 Poster Board II-416 It is generally assumed that T cell responses to HLA disparate targets arise from the naïve pool. However, the risk of graft-versus-host disease (GvHD) also increases if the donor has circulating T cells recognizing multiple persistent DNA viruses, suggesting that memory T cells may contribute to the overall alloresponse. We used a flow cytometric CFSE-based proliferation & activation (CD38 expression acquisition) assay to systematically assess the contribution of naïve and memory T cells to alloreactivity. In some experiments donor peripheral blood mononuclear cells (PBMC) were primed for 8 days with allogeneic, HLA mismatched PBMC (alloPBMC), after which the intracellular cytokine production (ICS) was examined upon rechallenge with autologous or allogeneic PBMC (alloPBMC). CFSE-labeled umbilical cord mononuclear cells (UCMC) from five donors were stimulated with an irradiated pool of HLA-disparate donor peripheral blood mononuclear cells (alloPBMC). By day 8, over 80% of the CD4+ and CD8+ T cells were CFSE[dim], i.e. had proliferated, and expressed the activation marker CD38, indicating that naïve T cells can mount an alloresponse. Next, PBMC from adult donors were separated into naïve (CD57-CD45RO-), memory (CD57-CD45RA- and CD57-CD62L- T cells), and effector (CD27-CD45RO-) T cells. We found that adult donor naïve T cells can also mount an alloresponse; importantly, both memory T cell subsets – and to a lesser extent, effector cells – were CFSE[dim]CD38+ at least to the same extent as naïve T cells, indicating their potential to respond to alloantigens in addition to their priming foreign antigen. These data were confirmed using a different approach: By first priming responder T cells with HLA-disparate donor PBMC for 8 days, followed by the enumeration of alloreactivity by ICS after stimulation with the original stimulator PBMC. To exclude reactivity of responder T cells with viral antigens present in PBMC (EBV; CMV), the experiment was repeated using activated T cells (T-APC) as antigen presenting cells, and yielded identical results. The direct ex vivo alloreactivity of T cell subsets was next tested by stimulating donor PBMC with HLA mismatched donor PBMC or activated T cells as APC in an 18 hour ICS. This experiment confirmed the rapid kinetics of alloreactivity and dominance of memory T cells in the alloresponse. Since a prominent role of memory T cells was apparent from our experiments we next tested T cells specific for DNA virus-derived antigens. EBV- and CMV-specific T cells, tested against a panel of 30 T-APC with a broad coverage of the most prominent HLA, displayed exquisite specificity for certain mismatched HLA alleles, indicating that DNA virus antigen-specific T cells may indeed cross-react with cellular antigens presented in the context of mismatched HLA class I and II proteins. Collectively our data demonstrate that both naïve and memory T cells mount an alloresponse, but that memory T cells are more rapidly and strongly recruited by alloantigen stimulation. These findings indicate that in man alloresponses are not confined to particular subsets of post-thymic T cells and that donor-derived viral antigen-specific T cells in an unrelated recipient may cross-react with peptide-MHC complexes against which they were not negatively selected. Disclosures: No relevant conflicts of interest to declare.

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Initiating antiretroviral treatment early in infancy has long-term benefits on the HIV reservoir in late childhood and adolescence

Clinical Infectious Diseases ◽

10.1093/cid/ciaa1931 ◽

2021 ◽

Author(s):

Véronique Avettand-Fenoel ◽

Jérôme Lechenadec ◽

Mariama Sadjo Diallo ◽

Marine Fillion ◽

Adeline Melard ◽

...

Keyword(s):

Mononuclear Cells ◽

Ex Vivo ◽

T Cell Subsets ◽

Effector Memory ◽

Childhood And Adolescence ◽

Older Children ◽

Peripheral Blood Mononuclear ◽

Hiv Reservoir ◽

Hiv Dna ◽

Cart Initiation

Abstract Background Early combined antiretroviral therapy (cART) limits the total HIV-DNA load in children. However, data on its impact in older children and adolescents remain scarce. This study aims to compare HIV reservoirs in children (5-12 years) and adolescents (13-17 years) who started cART before 6 months (early (E-)group) or after 2 years old (late (L-)group). Methods The ANRS-EP59-CLEAC study prospectively enrolled 76 HIV-1 perinatally-infected patients who reached HIV-RNA<400 copies/mL less than 24 months after cART initiation, regardless of subsequent viral suppression (E-group: 27 children, 9 adolescents; L-group: 19 children, 21 adolescents). Total and integrated HIV-DNA were quantified in blood and in CD4+ T cell subsets. A substudy assessed HIV reservoir inducibility after ex vivo peripheral blood mononuclear cells (PBMCs) stimulation. Results Total HIV-DNA levels were lower in early- than late-treated patients (Children: 2.14 vs 2.87 log cp/million PBMCs, p<0.0001; Adolescents: 2.25 vs 2.74log, p<0.0001). Low reservoir was independently associated with treatment precocity, protective HLA and low cumulative viremia since cART initiation. The 60 participants with undetectable integrated HIV-DNA started cART earlier than the other patients (4 vs 54 months, p=0.03). In those with sustained virological control, transitional memory and effector memory CD4+T cells were less infected in the E-group than in the L-group (p=0.03 and 0.02, respectively). Viral inducibility of reservoir cells after normalization to HIV-DNA levels was similar between the groups. Conclusions Early cART results in a smaller blood HIV reservoir until adolescence, but all tested participants had an inducible reservoir. This deserves cautious consideration for HIV remission strategies.

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The Repertoires of Circulating Human CD8+ Central and Effector Memory T Cell Subsets Are Largely Distinct

Immunity ◽

10.1016/s1074-7613(03)00020-7 ◽

2003 ◽

Vol 18 (2) ◽

pp. 193-204 ◽

Cited By ~ 116

Author(s):

Véronique Baron ◽

Cécile Bouneaud ◽

Ana Cumano ◽

Annick Lim ◽

T.Petteri Arstila ◽

...

Keyword(s):

T Cell ◽

T Cell Subsets ◽

Effector Memory ◽

Memory T Cell

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CD4+ T cells are essential for the development of destructive thyroiditis induced by anti–PD-1 antibody in thyroglobulin-immunized mice

Science Translational Medicine ◽

10.1126/scitranslmed.abb7495 ◽

2021 ◽

Vol 13 (593) ◽

pp. eabb7495

Author(s):

Yoshinori Yasuda ◽

Shintaro Iwama ◽

Daisuke Sugiyama ◽

Takayuki Okuji ◽

Tomoko Kobayashi ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Lymph Nodes ◽

Mononuclear Cells ◽

Critical Role ◽

T Cell Subsets ◽

Effector Memory ◽

Histopathological Analysis ◽

Activated T Cells ◽

Cervical Lymph Nodes

Immune-related adverse events induced by anti–programmed cell death–1 antibodies (PD-1-Ab), including destructive thyroiditis (thyroid-irAE), are thought to be caused by activated T cells. However, the T cell subsets that are directly responsible for damaging self-organs remain unclear. To clarify which T cell subsets are involved in the development of thyroid-irAE, a mouse model of thyroid-irAE was analyzed. PD-1-Ab administration 2.5 months after immunization with thyroglobulin caused destructive thyroiditis. Thyroiditis was completely prevented by previous depletion of CD4+ T cells and partially prevented by depleting CD8+ T cells. The frequencies of central and effector memory CD4+ T cell subsets and the secretion of interferon-γ after stimulation with thyroglobulin were increased in the cervical lymph nodes of mice with thyroid-irAE compared with controls. Histopathological analysis revealed infiltration of CD4+ T cells expressing granzyme B in thyroid glands and major histocompatibility complex class II expression on thyrocytes in mice with thyroid-irAE. Adoptive transfer of CD4+ T cells from cervical lymph nodes in mice with thyroid-irAE caused destruction of thyroid follicular architecture in the irradiated recipient mice. Flow cytometric analyses showed that the frequencies of central and effector memory CD4+ T cells expressing the cytotoxic marker CD27 were higher in peripheral blood mononuclear cells collected from patients with thyroid-irAE induced by PD-1-Ab versus those without. These data suggest a critical role for cytotoxic memory CD4+ T cells activated by PD-1-Ab in the pathogenesis of thyroid-irAE.

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Reconstitution of the peripheral immune repertoire following withdrawal of fingolimod

Multiple Sclerosis Journal ◽

10.1177/1352458517713147 ◽

2017 ◽

Vol 23 (9) ◽

pp. 1225-1232 ◽

Cited By ~ 17

Author(s):

Mahtab Ghadiri ◽

Leslie Fitz-Gerald ◽

Ayman Rezk ◽

Rui Li ◽

Mukanthu Nyirenda ◽

...

Keyword(s):

T Cell ◽

Clinical Laboratory ◽

Alternative Therapies ◽

T Cell Subsets ◽

Effector Memory ◽

Immune Repertoire ◽

Drug Induced ◽

Memory T Cell ◽

Cell Counts ◽

Pre Treatment

Background: Following fingolimod cessation, immune reconstitution or lack thereof may have consequences for disease rebound or safety of commencing alternative therapies. Objective: To examine the degree and profile of peripheral blood lymphocyte reconstitution following fingolimod withdrawal. Methods: Total lymphocyte counts (TLC) and CD4+/CD8+ T-cell counts were measured in 18 multiple sclerosis (MS) patients pre-treatment, on fingolimod, and up to 8–9 months post-cessation. T-cell subsets were analyzed using flow cytometry. Results: At 2-week post-fingolimod cessation, TLC reconstitution was variable and not correlated with age, treatment duration, pre-, or on-treatment TLC. Despite normalization of TLC and CD4+:CD8+ ratios over months, naive subsets remained lower and effector memory subsets higher in frequency compared with pre-treatment. Drug-induced increases in ratios of regulatory to pathogenic Th17-containing central memory populations appeared to rapidly return to baseline. Conclusion: Early peripheral lymphocyte reconstitution after fingolimod withdrawal remains partial and heterogeneous. Relative frequencies of circulating naive and memory T-cell subsets may not recover for many months, even when clinical laboratory tests have normalized. Analyzing specific components of the peripheral immune repertoire helps define the overall immune status of patients. To be determined is whether assessment of such immune measures will have implications for the timing and safety of commencing alternative therapies.

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Proliferation and differentiation potential of human CD8+ memory T-cell subsets in response to antigen or homeostatic cytokines

Blood ◽

10.1182/blood-2002-11-3577 ◽

2003 ◽

Vol 101 (11) ◽

pp. 4260-4266 ◽

Cited By ~ 361

Author(s):

Jens Geginat ◽

Antonio Lanzavecchia ◽

Federica Sallusto

Keyword(s):

T Cell ◽

Receptor Expression ◽

T Cell Subsets ◽

Brdu Incorporation ◽

Effector Memory ◽

Differentiation Potential ◽

Memory T Cell ◽

Proliferation And Differentiation ◽

Homeostatic Cytokines

Abstract Four human CD8+ T-cell subsets, naive (CCR7+CD45RA+), central memory (TCM, CCR7+CD45RA–), effector memory (TEM, CCR7–CD45RA–), and CD45RA+ effector memory cells (TEMRA, CCR7–CD45RA+) were compared for their capacity to proliferate and differentiate in response to antigen or homeostatic cytokines. Cytokine responsiveness and interleukin-15 receptor expression were low in naive T cells and progressively increased from TCM to TEM and TEMRA. In contrast, the capacity to accumulate in response to T-cell receptor (TCR) or cytokine stimulation showed a reciprocal pattern and was associated with resistance to cell death and Bcl-2 expression. Whereas all TCR-stimulated cells acquired a CD45RA–CCR7– phenotype, cytokine-stimulated cells maintained their phenotype with the exception of TCM cells, which expressed CCR7, CD45RA, and perforin in various combinations. Single CD8+ TCM cells, but not TEM cells, could be expanded with cytokines, and the obtained clones displayed several distinct phenotypes, suggesting that TCM cells are heterogeneous. Consistently, CCR4 expression in the CD8+ TCM pool discriminated CCR4+ type 2 polarized cells (Tc2) and CCR4–CTL precursors. Finally, ex vivo bromodeoxyuridine (BrdU) incorporation experiments revealed that memory subsets have different in vivo proliferation rates, with CCR4–TCM having the highest turnover and TEMRA the lowest. These results show that human CD8+ memory T-cell subsets have different proliferation and differentiation potentials in vitro and in vivo. Furthermore, they suggest that TEMRA cells are generated from a TCM subset upon homeostatic proliferation in the absence of antigen.

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Application of a Standardized Flow Cytometry Panel for Defining and Monitoring the Immunophenotype of CAR-T Cells

Blood ◽

10.1182/blood-2019-128745 ◽

2019 ◽

Vol 134 (Supplement_1) ◽

pp. 5626-5626

Author(s):

Irene Scarfò ◽

Kathleen Gallagher ◽

Marcela V. Maus ◽

Rebecca Larson ◽

Maegan Sheehan ◽

...

Keyword(s):

T Cells ◽

Stem Cell ◽

T Cell ◽

Unmet Need ◽

Central Memory ◽

T Cell Subsets ◽

Effector Memory ◽

Cell Phenotype ◽

Memory T Cell ◽

Car T

Chimeric antigen receptor T-cells (CAR-T) have emerged as an extremely promising therapy for hematological malignancies. The immunophenotype of apheresis material and the CAR-T cell product is known to be predictive of the likelihood of response to treatment of certain malignancies. Central memory and stem cell-like memory T cell phenotypes are associated with a more sustained proliferative response and long-term CAR-T persistence (Fraietta et al, Nature Medicine, 2018). There is an unmet need for standardized methods and reagents to reliably profile the memory phenotype of CAR-Ts to better evaluate product quality, and support improvements in CAR-T manufacturing. The BD Biosciences dried memory T-cell panel contains a pre-validated mixture of 7 antibodies for the identification of naïve, stem cell memory, central memory and effector memory CD4+ and CD8+ T cell subsets. The pre-mixed dried antibody tube offers consistency in staining profiles over time and reduces the risk of operator errors. Additional drop-in antibodies can complement the panel and enable more in-depth evaluation of the T cell phenotype. Here we demonstrate the use of this panel with drop-in markers to monitor changes in expression of PD-1, TIM-3, LAG-3, HLA-DR, CD45RO, and CXCR3 on T cells transduced to express our novel anti-CD37 CAR. Cells were stained at day 0 prior to transduction, day 7, and following resting and re-stimulation, and acquired on a 12 color BD FACS Lyric. The use of a standardized memory T-cell panel will allow us to more accurately evaluate how T-cell phenotype impacts on the efficacy and longevity of response in patients receiving CAR-T therapies. Disclosures Maus: INFO PENDING: Other: INFO PENDING. Bornheimer:BD Biosciences: Employment. Hanley:BD Biosciences: Employment. Frigault:Novartis: Patents & Royalties: Royalty; Arcellx, Celgene, Foundation Medicine, Kite/Gilead, Nkarta, Novartis, and Xenetic: Consultancy.

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The differential immune responses to COVID-19 in peripheral and lung revealed by single-cell RNA sequencing

10.1101/2020.08.15.20175638 ◽

2020 ◽

Author(s):

Gang Xu ◽

Furong Qi ◽

Hanjie Li ◽

Qianting Yang ◽

Haiyan Wang ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Single Cell ◽

Mononuclear Cells ◽

Immune Cell ◽

Close Association ◽

Suppressor Cells ◽

T Cell Subsets ◽

Peripheral Blood Mononuclear ◽

Peripheral T Cells

Understanding the mechanism that leads to immune dysfunction induced by SARS-CoV2 virus is crucial to develop treatment for severe COVID-19. Here, using single cell RNA-seq, we characterized the peripheral blood mononuclear cells (PBMC) from uninfected controls and COVID-19 patients, and cells in paired broncho-alveolar lavage fluid (BALF). We found a close association of decreased dendritic cells (DC) and increased monocytes resembling myeloid-derived suppressor cells (MDSC) which correlated with lymphopenia and inflammation in the blood of severe COVID-19 patients. Those MDSC-like monocytes were immune-paralyzed. In contrast, monocyte-macrophages in BALFs of COVID-19 patients produced massive amounts of cytokines and chemokines, but secreted little interferons. The frequencies of peripheral T cells and NK cells were significantly decreased in severe COVID-19 patients, especially for innate-like T and various CD8+ T cell subsets, compared to health controls. In contrast, the proportions of various activated CD4+ T cell subsets, including Th1, Th2 and Th17-like cells were increased and more clonally expanded in severe COVID-19 patients. Patients' peripheral T cells showed no sign of exhaustion or augmented cell death, whereas T cells in BALFs produced higher levels of IFNG, TNF, CCL4 and CCL5 etc. Paired TCR tracking indicated abundant recruitment of peripheral T cells to the patients' lung. Together, this study comprehensively depicts how the immune cell landscape is perturbed in severe COVID-19.

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