Journal: Physiological Reports

Article Title: Sex differences in circulating platelet‐derived CD41 + extracellular vesicles in healthy adults

doi: 10.14814/phy2.70932

Figure Lengend Snippet: Nano flow cytometry measures of MV phenotype in males and females. (a) CD3+ lymphocyte‐derived MVs. (b) CD14+ monocyte‐derived MVs. (c) CD16+ neutrophil‐derived MVs. (d) CD45+ leukocyte‐derived MVs. (e) CD31+ endothelial cell‐derived MVs. (f) CD62E+ endothelial activation‐derived MVs. (g) CD41+ platelet‐derived MVs. (h) MVs expressing Annexin A5. Individual data points are presented as MV counts/μL. Clear circles represent females and dark filled circles represent males. Statistical comparisons between sexes were performed using linear models adjusted for age and BMI. Data are presented as untransformed values for visualization following outlier removal using a 3*SD criterion. Statistical analyses were performed on log‐transformed data where appropriate.

Article Snippet: Panel 2 received 2 μL each of CD3 PE (130‐114‐519, Miltenyi Biotec), CD14 PerCP‐Vio700 (130‐110‐523), CD16 PE‐Vio615 (130‐119‐995), CD31 FITC (130‐110‐668), CD41 APC (130‐123‐301), and 10 μL of CD62E PE (130‐104‐643).

Techniques: Flow Cytometry, Derivative Assay, Activation Assay, Expressing, Transformation Assay

Journal: Nature Medicine

Article Title: Multi-antigen-targeting T cells in pediatric central nervous system tumors: a phase 1 trial

doi: 10.1038/s41591-026-04449-9

Figure Lengend Snippet: Flow immunophenotyping release testing data are presented in A-G and IFNγ ELISpot data presented in H-K (Arm A: n = 12; Arms B/C: n = 27). Each data point represents a separate TAA-T cell product infused into patients (biological replicates). Lower whisker is defined as the minima, upper whisker is defined as the maxima, center is defined as median, lower bound of box is defined as Q1 (25th percentile), and upper bound of box is defined as Q3 (75th percentile). a ) % Total T cells (CD3 + ). Arm A: Minima=51, Maxima=99, Median=97, 25th Percentile=95, 75th Percentile=99; Arms B/C: Minima=31, Maxima=100, Median=97, 25th Percentile=94, 75th Percentile=98. b ) % CD4 + T cells (CD3 + CD4+ of CD45 + ). Arm A: Minima=0.50, Maxima=88, Median=16, 25th Percentile=2.6, 75th Percentile=38; Arms B/C: Minima=0.40, Maxima=55, Median=9.9, 25th Percentile=6.1, 75th Percentile=17. c ) %CD8 + T cells (% CD3 + CD8+ of CD45 + ). Arm A: Minima=4.3, Maxima=88, Median=37, 25th Percentile=22, 75th Percentile=47; Arms B/C: Minima=19, Maxima=91, Median=51, 25th Percentile=36, 75th Percentile=71. d ) % αβ T cells (% TCRαβ+ of CD3 + ). Arm A: Minima=8.2, Maxima=94, Median=69, 25th Percentile=28, 75th Percentile=86; Arms B/C: Minima=32, Maxima=98, Median=71, 25th Percentile=48, 75th Percentile=84. e ) % γδ T cells (% TCRγδ+ of CD3 + ). Arm A: Minima=5.1, Maxima=79, Median=28, 25th Percentile=8.4, 75th Percentile=67; Arms B/C: Minima=1.1, Maxima=83, Median=25, 25th Percentile=10, 75th Percentile=53. f ) % CD3 + CD16 + CD56+ of CD45 + . Arm A: Minima=2.8, Maxima=25, Median=11, 25th Percentile=7.2, 75th Percentile=17; Arms B/C: Minima=3.4, Maxima=64, Median=16, 25th Percentile=8.1, 75th Percentile=28. g ) % NK cells (CD16/CD56 + CD3- of CD45 + ). Arm A: Minima=0.40, Maxima=48, Median=1.0, 25th Percentile=0.48, 75th Percentile=4.6; Arms B/C: Minima=0.10, Maxima=62, Median=1.6, 25th Percentile=0.60, 75th Percentile=6.1. h - i ) IFNγ ELISpot with background correction for Arm A ( h ) and Arms B/C ( i ). Negative values after background normalization have been normalized to zero for visual representation.

Article Snippet: Antibodies for the T/NK panel included CD8 FITC (Miltenyi Biotec, cat. no. 130-110-677), CD16 PE (Miltenyi Biotec, cat. no. 130-113-393), CD56 PE (Miltenyi Biotec, cat. no. 130-113-312), CD3 Per CP Vio-770 (Miltenyi Biotec, cat. no. 130-113-141), CD4 PE-Vio-770 (Miltenyi Biotec, cat. no. 130-113-227) and CD45 APC (Miltenyi Biotec, cat. no. 130-110-633).

Techniques: Enzyme-linked Immunospot, Whisker Assay

Journal: International Journal of Molecular Sciences

Article Title: Proinflammatory Cytokine Preconditioning Enhances the Therapeutic Potency of Different Types of MSCs in Inflammation

doi: 10.3390/ijms27094090

Figure Lengend Snippet: The effects of naïve and Cytomix-stimulated MSCs on T cell proliferation. The gating strategy is shown in . ( A – C ) BM-MSCs, iMSCs WT and iMSCs B2M KO inhibition on the expansion of CD3+, CD3+CD4+ and CD3+CD8+ T cells via CM. Cytomix enhanced the three types of MSCs’ inhibition on the EI of T cells. ( D – F ) All types of MSCs significantly inhibited the division of CD3+, CD3+CD4+ and CD3+CD8+ T cells when co-cultured with PBMCs, in a dose-dependent manner. ( G – I ) All of the MSCs, naïve or Cytomix stimulated, significantly reduced the EI of CD3+, CD3+CD4+ or CD3+CD8+ T cells, at the lowest ratio of MSCs, with iMSCs WT both naïve and Cytomix-stimulated presenting the strongest inhibition. ( J ) Co-culturing all types of MSCs, naïve or Cytomix-stimulated, increased the production of G-CSF in the media, with a dose-dependent effect. ( K ) All types of MSCs significantly decreased the level of IFN-γ at the ratios of 1:20 and 1:10, while the three types of naïve MSCs significantly decreased it at 1:50. ( L ) Naïve and Cytomix-stimulated iMSC WTs significantly increased the level of TNF-α at the ratios of 1:50, 1:20 and 1:10, while Cytomix-stimulated iMSC B2M KO significantly increased it at the ratio of 1:50 and 1:20. ( M ) Naïve BM-MSCs significantly increased the level of IL-10 production at the ratios of 1:20 and 1:10. Representative results of four independent experiments are shown. Results are shown as mean ± SD ( n = 3 in CM; n = 4 in co-culture, each group), and analysed by two-way ANOVA with Šídák’s multiple comparisons test to compare naïve vs. Cytomix in each type of MSCs (no significance in all of the settings), or Tukey’s multiple comparisons test to compare different ratios of MSCs to PBMCs in each MSC group, or Dunnett’s multiple comparisons test to compare different MSC groups vs. vehicle in the setting of the same ratio. */**/***/**** represent comparisons of each type of MSC vs. vehicle; #/## represent comparisons of different ratios of each cell type with different levels of p < 0.05/0.01. EI: expansion index; DP: division percentage.

Article Snippet: The PBMCs were stained with anti-human CD3-APC-Vio770, CD4-APC and CD8-PE-Vio770 antibodies (Cat# 130-113-136, 130-113-222, 130-110-680; Miltenyi Biotec) for 30 min on ice in the dark.

Techniques: Inhibition, Cell Culture, Co-Culture Assay