Journal: Cell Reports Medicine

Article Title: Delayed viral rebound post-ART interruption in infant macaques given SIV-specific neutralizing antibodies

doi: 10.1016/j.xcrm.2026.102636

Figure Lengend Snippet: SIV-specific T cell responses and correlation with viral rebound AUC (A) SIVgag-specific CD8 + T cell responses were quantified by AIM assay, using Boolean gating to show the frequencies of CD69 + CD25 + , CD69 + OX40 + , or CD25 + OX40 + cells following stimulation with an SIVgag peptide pool after background subtraction. The pre-ATI (and pre-RhmAb for that group) time point was compared to the rebound peak post-ATI. Nonparametric Wilcoxon matched-pairs signed rank test was used for statistics. N = 4 and N = 5 for controls at each time point based on sample availability. (B) Graph shows the frequency of SIVgag-specific CD8 + T cells at peak rebound in the RhmAb group and rebound viral load AUC. Statistical analysis was performed by calculating the nonparametric Spearman’s rank correlation coefficient (r) that was compared to zero.

Article Snippet: PE-Vio770 Mouse Anti-Human CD25 , Miltenyi Biotec , 130-113-287; RRID:AB_2784092.

Techniques:

Journal: bioRxiv

Article Title: Coordinated IL-2 and TGF-β Signaling via a Novel Fusion Protein Selectively Expands and Activates Regulatory T Cells

doi: 10.64898/2026.03.09.709648

Figure Lengend Snippet: ( A ). Flow cytometric analysis shows the percentage of CD4 + Foxp3 + expressing cells following treatment of mice with PBS or CUE-401. CD4⁺Foxp3⁺ Tregs were identified, and UMAP projections were generated using OMIQ software to cluster cells based on protein expression. The feature plot shows CD25 expression within the Foxp3⁺ populations. ( B ). Cells were gated on the CD4⁺Foxp3⁺ populations to assess surface expression of GITR, CD25, CTLA-4 and CD103. ( C ). Histograms display the expression of GITR, CD25, CTLA-4, and CD103 within the Treg compartment of PBS-treated controls compared to CUE-401–treated mice. ( D ). Heatmap from bulk RNA sequencing of sorted Tregs, highlighting differentially expressed genes between PBS and CUE-401 treated mice. Genes included had a log₂ fold change > 2 and adjusted p-value < 0.05, using the Benjamini–Hochberg correction to control false discovery rate (FDR). For all flow cytometry plots, data are representative of individual spleens from a single experiment with 3 mice per treatment group. PBS-treated, age-matched mice served as controls in all analyses.

Article Snippet: To deplete CD25 + cells, cells were incubated for 10 min with anti-CD25-PE (PC61), washed in MACS buffer, and incubated for an additional 10 min with anti-PE MACS microbeads (Miltenyi Biotec, Bergisch Gladbach) then subjected to autoMACS purification (Miltenyi Biotec).

Techniques: Expressing, Generated, Software, RNA Sequencing, Control, Flow Cytometry

Journal: bioRxiv

Article Title: Coordinated IL-2 and TGF-β Signaling via a Novel Fusion Protein Selectively Expands and Activates Regulatory T Cells

doi: 10.64898/2026.03.09.709648

Figure Lengend Snippet: (A). Representative flow cytometric analysis showing the percentage of CD4 + gated cells expressing Foxp3 six days after treatment with PBS (left), CUE-401ΔTGF-β3 (middle), or CUE-401 (right) in vivo. (B). Combined data showing the percentages of CD4 + Foxp3 + Tregs in PBS, CUE-401ΔTGF-β3, or CUE-401 treated mice. ( C ). Representative flow cytometric analysis comparing GITR by CD38 expression in CD4 + Foxp3 + Tregs following the indicated treatments. ( D ). Combined data showing the percentage of GITR + CD38 + Tregs following PBS, CUE-401ΔTGF-β3, or CUE-401 treatment. ( E ). Representative flow cytometric analysis comparing CD25 by CD103 of CD4 + Foxp3 - gated cells. ( F ). Combined data showing the percentage of CD25 + CD4 + Foxp3 - cells and CD103 + CD4 + Foxp3 - cells six days after treatment. Data shown are representative of two independent experiments (N = 9 per group). Bars represent the median ± 95% confidence interval. ( G , H ). Heatmaps of gene expression profiles in sorted CD4⁺Foxp3.eGFP⁺ Treg cells or CD4 + Foxp3.eGFP - Tconv cells, derived from a subset of control or treated mice (N = 2,3 per group, respectively). P values for flow results were determined utilizing an ordinary one-way ANOVA with Tukey’s multiple comparisons test with * p = 0.0332, ** p = 0.0021, *** p = 0.0002, and **** p < 0.0001.

Article Snippet: To deplete CD25 + cells, cells were incubated for 10 min with anti-CD25-PE (PC61), washed in MACS buffer, and incubated for an additional 10 min with anti-PE MACS microbeads (Miltenyi Biotec, Bergisch Gladbach) then subjected to autoMACS purification (Miltenyi Biotec).

Techniques: Expressing, In Vivo, Gene Expression, Derivative Assay, Control

Journal: bioRxiv

Article Title: Coordinated IL-2 and TGF-β Signaling via a Novel Fusion Protein Selectively Expands and Activates Regulatory T Cells

doi: 10.64898/2026.03.09.709648

Figure Lengend Snippet: (A). Representative flow cytometric analysis of CD4 gated Foxp3 + cells in PBS and CUE-401 treated BALB/c nude mice 7 days after splenocyte transfer. ( B ). Cumulative data showing the percentages of CD4 + Foxp3 + Tregs in PBS and CUE-401 treated mice 6 days after a single treatment. ( C ). CD4⁺Foxp3⁺ Tregs were identified in each treatment condition, and UMAP projections generated distinct clusters of cells based on protein expression. ( D ). Representative flow cytometric analysis of CD4 + Foxp3 + gated, CD25 expressing cells. ( E ). Combined data showing the percentages of CD25 expressing Tregs in PBS and CUE-401 treated mice. ( F ). Single cell RNA sequencing clustering analysis of T cells. T cell populations were subset and Foxp3 expression identified the Treg populations in PBS and CUE-401 treated mice. ( G ). Foxp3 expressing T cells were re-clustered and signatures were generated based on IL-2/STAT5 signaling transcripts that were identified through gene set enrichment analysis ( H ). Heatmap that highlights differentially expressed genes in single Foxp3 + cells between PBS and CUE-401 treatment. Differential expression analysis between CUE-401 and PBS Tregs used the Wilcoxon rank-sum test implemented in Seurat. P-values were adjusted for multiple comparisons using the Benjamini–Hochberg method to control false discovery rate (FDR). Representative images in (A) from 3 independent experiments with 3-5 age matched female mice per experiment. P -values were determined utilizing Mann-Whitney U test with p < 0.05 as significant for the flow cytometry analyses. Single cell analysis in (F-H) generated cells of 8 total mice in 2 independent experiments (4 treated and 4 untreated).

Article Snippet: To deplete CD25 + cells, cells were incubated for 10 min with anti-CD25-PE (PC61), washed in MACS buffer, and incubated for an additional 10 min with anti-PE MACS microbeads (Miltenyi Biotec, Bergisch Gladbach) then subjected to autoMACS purification (Miltenyi Biotec).

Techniques: Generated, Expressing, Single Cell, RNA Sequencing, Quantitative Proteomics, Control, MANN-WHITNEY, Flow Cytometry, Single-cell Analysis