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Schematic representation of the main altered metabolic pathways in Treg cel...
Published: 15 June 2024
FIGURE 1. Schematic representation of the main altered metabolic pathways in Treg cells from multiple sclerosis (MS) subjects. Treg cell from MS subjects (right) is characterized by different metabolic alterations as compared with a Treg cell from healthy subjects (left), such as 1) hyperactivatio... More about this image found in Schematic representation of the main altered metabolic pathways in Treg cel...
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Schematic representation of the main Treg cell metabolic alterations in sys...
Published: 15 June 2024
FIGURE 2. Schematic representation of the main Treg cell metabolic alterations in systemic lupus erythematosus. (1) Treg cells from systemic lupus erythematosus (SLE) subjects are characterized by giant mitochondria, increased generation of oxidation products, and enhanced ATP consumption associated with DNA damage and Treg cell death; (2) inhibition of calcium/calmodulin-dependent protein kinase 4 (CaMK4) enhances Treg cell immunosuppressive function through modulation of intracellular metabolism; and (3) conditional deletion of the SLE susceptibility gene Esrrg in Treg cells results in increased ROS generation, impaired oxygen consumption and ATP and NAD+ production, reduced expression of IL2ra and Trbc1, and impaired TGF-β signaling and increased mTOR activation. More about this image found in Schematic representation of the main Treg cell metabolic alterations in sys...
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Biphasic decline of VV-specific Abs after revaccination.   ( A ) We analyze...
Published: 12 June 2024
FIGURE 1. Biphasic decline of VV-specific Abs after revaccination. ( A ) We analyzed the kinetics of Ab titers in four long-term blood donors from subjects 1–4 shown in Fig. 2 of Amanna et al. ( 8 ). These individuals were followed up for 20+ years during which they had been revaccinated with VV. ( B ) We fitted a mathematical model of humoral immune response (Eq. 1 ) to subsets of the data that include revaccination and estimated the rate of Ab expansion (ρ), the proportion of Ab conversion into a long-lived population (fm), and the half-life of the humoral immunity (T1/2, B and see Table I for estimated model parameters). Supplemental Fig. 1C – 1F shows dynamics of individual subpopulations of ASCs predicted by the model. ( C ) Kinetics of Ab response following VV revaccination in one volunteer (donor 97) suggests infinite half-life of the long-term memory [see main text for best fit parameters; these data were published previously ( 10 )]. In (B) and (C), markers denote the data, and lines are predictions of the mathematical model. ( D ) Sparse measurements of Ab titers after vaccination allow alternative mathematical models (Eq. 1 with different parameter sets) with drastically different predicted longevities of Abs. Here, the markers are average Ab titers from Fig. 1Ciii of Arunachalam et al. ( 4 ), and lines are predictions of two alternative mathematical models with different assumed subpopulations of ASCs. More about this image found in Biphasic decline of VV-specific Abs after revaccination. ( A ) We analyze...
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The approach to stable Ab levels takes months to years after revaccination ...
Published: 12 June 2024
FIGURE 2. The approach to stable Ab levels takes months to years after revaccination with VV or tetanus vaccine. We fitted several alternative mathematical models for Ab decay (Eq. 2 ) to data by varying the number of subpopulations for four volunteers revaccinated with VV ( A ) or tetanus ( B ) vaccines. In the analysis, we included only the data from the peak Ab response at the first instance after an apparent revaccination (noted by the horizontal dashed line). In both cases, the models with two subpopulations fitted the data better than the model with a single subpopulation, but a larger model (n = 3) did not improve the model fit quality but did provide longer half-life times for persisting Abs. For each fit, we provide the estimated half-life time (T1/2) based on the slowest decay rate. In both panels, markers denote the data, and lines are predictions of the mathematical model. Parameters of the best fit models are shown in Table II . Note the different scales of the follow-up for boosting with two different vaccines. EU, ELISA units. More about this image found in The approach to stable Ab levels takes months to years after revaccination ...
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Fold expansion, viability, and transduction efficiency.   ( A ) Schematic o...
Published: 12 June 2024
FIGURE 1. Fold expansion, viability, and transduction efficiency. ( A ) Schematic of 7-d culture used in this study. ( B ) Fold expansion for each day of culture was calculated by dividing the initial total nucleated cells (TNC) on the subsequent day by the final TNCs on the previous day. Untransduced and transduced groups are shown for all donors. ( C ) Viability is shown for day 7 for all groups and donors. ( D ) A vector MOI of 80 was used for donor C compared with 40 for donors A and B. For all sections, *p ≤ 0.05. More about this image found in Fold expansion, viability, and transduction efficiency. ( A ) Schematic o...
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T cell activation and exhaustion markers.   ( A ) Percent CD25<sup>+</sup>,...
Published: 12 June 2024
FIGURE 2. T cell activation and exhaustion markers. ( A ) Percent CD25+, PD1+, LAG3+ of CD3+ cells by flow cytometry is shown for the duration of culture for all transduced groups and the unactivated control group. Dotted line is shown at 50% for ease of viewing differences in PD1 and LAG3 expression. ( B ) Averages from (A) are shown overlaid for each day. Significant differences are shown for PD1+% on day 2 and LAG3+% on days 2 and 4. For all sections, *p ≤ 0.05. More about this image found in T cell activation and exhaustion markers. ( A ) Percent CD25+,...
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Effects of activators on T cell subsets.   ( A ) CD4/CD8 ratio was calculat...
Published: 12 June 2024
FIGURE 3. Effects of activators on T cell subsets. ( A ) CD4/CD8 ratio was calculated for all donors and groups on days 0 and 7 by dividing percent CD4+ by CD8+ of CD3+. ( B ) Day 7 frequencies of CD4+ (left) and CD8+ (right) of CD3+ cells are shown. ( C ) Frequencies of cells expressing markers associated with naive (upper left), central memory (upper right), effector memory (lower left), or T effector (lower right) cell populations on day 7. For all sections, *p ≤ 0.05, **p ≤ 0.01. More about this image found in Effects of activators on T cell subsets. ( A ) CD4/CD8 ratio was calculat...
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T cell functional assays.   ( A ) The tumor cell viable total nucleated cel...
Published: 12 June 2024
FIGURE 4. T cell functional assays. ( A ) The tumor cell viable total nucleated cells (TNCs) are shown at the 24-h time point at an E:T cell ratio of 1:1 (left). The seeding tumor cell TNCs are represented with the dotted line at 5 × 104. At the 48-h time point at 1:1 E:T, very few tumor cells remained (right). ( B ) Cytokine protein concentrations were analyzed using the Bio-Plex 200 for 1:1 E:T ratio killing assays. More about this image found in T cell functional assays. ( A ) The tumor cell viable total nucleated cel...
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Gene expression profile using the CAR T characterization panel from NanoStr...
Published: 12 June 2024
FIGURE 5. Gene expression profile using the CAR T characterization panel from NanoString nCounter. ( A ) Pathways are listed on the y-axis, and activator groups are listed on the x-axis. Yellow indicates high scores; black indicates low scores. Scores are displayed on the same scale via a Z-transformation. ( B ) Pathways from (A) with significant differences on the pathway scores among four activator groups. x-Axis represents pathways; y-axis indicates pathway score. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. ( C ) Principal-component analysis loading plot indicates the distribution of the analyzed genes for each donor/method pair. Percentages represent variance captured by each principal component, 1 and 2. Controls are shown as ▲ (nonstimulated) and + (day 0), and donors are colored differently. ( D ) Gene expression data of 780 genes are shown as a heatmap for each sample. Vertical axis represents differentially expressed genes (DEGs) with log2 gene expression intensity values. Red and green represent upregulated and downregulated genes, respectively. Heatmap is clustered using both gene categories and samples (methods), and gene expression data were scaled by each sample. More about this image found in Gene expression profile using the CAR T characterization panel from NanoStr...
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