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: Science Advances

Article Title: Harnessing skin-resident γδ T cells for immunotherapy in cutaneous squamous cell carcinoma

doi: 10.1126/sciadv.aec7215

Figure Lengend Snippet: ( A ) Expression of indicated chemokine receptors by in vitro expanded, live gated CD3 + Vδ1 + γδ T cells. Blood-derived αβ T cells were used as staining control. ( B ) As (A), graphical summary of the percentage of Vδ1 + γδ T cell expressing the chemokine receptors ( n = 5 skin donors). Error bars represent mean ± SD. ( C ) 10 × 10 6 human skin–derived T cells, containing approximately 7% of Vδ1 + γδ T cells, were injected intravenously (i.v.) into NSG mice carrying a xSCC of a volume ranging from 100 to 200 mm 3 [reached approximately 60 to 80 days (d) post–i.d. injection of SCC-13 cells]. Each mouse was injected intraperitoneally (i.p.) with recombinant IL-2 and IL-15 daily until the harvest day. Figure created in BioRender. I. Gratz (2026) https://BioRender.com/zmnd28b . ( D ) Representative plot of the percentage of Vδ1 + γδ T cells engrafted in the spleen, blood, xSCC and murine skin 2, 7, or 14 days posttransfer. ( E to G ) Bar graphs show the absolute numbers of ingoing Vδ1 + γδ T cells, Vδ1 + γδ T cells engrafting spleen and blood normalized to mouse weight (grams), and xSCC and murine skin normalized to tissue weight (grams). (E) n = 7 mice per group; pool of two independent experiments. (F) n = 12 mice per group; pool of four independent experiments; (G) n = 12 mice per group; pool of two independent experiments. Each symbol represents one skin donor. Error bars represent mean ± SD. Statistical significance was determined using the Kruskal-Wallis test with Dunn’s multiple comparisons test. All data points, including extreme values, are shown. ( H ) Representative immunofluorescent staining of colocalized TCRδ/DAPI in HD skin and xSCC 7 days after γδ transfer. Scale bars, 100 μm. Staining controls are shown in fig. S4.

Article Snippet: Human skin–derived T lymphocytes were labeled with Cell Proliferation Dye eFluor 450 (10 μM, Thermo Fisher Scientific, catalog no. 65- 0842-85), washed with PBS, and subsequently stimulated for 6 days with rhuIL-2 (100 IU/ml; BioLegend, catalog no. 589108), rhuIL-15 (20 ng/ml; BioLegend, catalog no. 570603), in addition with mouse anti-human CD3 monoclonal antibody (1 μg/ml; Miltenyi Biotec, catalog no. 130-093-387, RRID:AB_1036144) alone or with rhuIL-1α (9 ng/ml; BioLegend, catalog no. 570004) and rhuIL-18 (9 ng/ml, BioLegend, catalog no. 592102) applied individually or in combination in cTexMacs medium.

Techniques: Expressing, In Vitro, Derivative Assay, Staining, Control, Injection, Recombinant

Journal: Science Advances

Article Title: Harnessing skin-resident γδ T cells for immunotherapy in cutaneous squamous cell carcinoma

doi: 10.1126/sciadv.aec7215

Figure Lengend Snippet: ( A ) Levels of cytokines (pg/mg tissue) produced by HD skin, huSCC, ES, and xSCC. Heatmap bars represent the mean of n = 5 HD skin and huSCC donors, and mean n = 5 of xenograft mice. ( B ) Representative gating strategy and bar graphs of the human skin–derived and ex vivo expanded Vδ1 + γδ T cells expressing IL-1RAcP and IL-18Rα. Peripheral blood αβ T cells and ex vivo expanded skin-derived αβ T cells were used as staining controls. Mean of n = 5 skin donors. ( C ) eFluor450-labeled γδ T cells were cultured under basal conditions [unstimulated or with IL-2 (100 IU/ml) and IL-15 (20 ng/ml)] or stimulated with anti-CD3 (1 μg/ml) and/or IL-1α and IL-18 (9 ng/ml) for 6 days. Proliferation was assessed by the median fluorescence intensity (MFI) of eFluor450 in Vδ1 + by flow cytometry. The representative histograms show the eFluor450 dilution in Vδ1 + T cells in the different conditions. Cell counts were normalized to unit area. ( D ) Bar graphs show the fold change of efluor450 MFI of Vδ1 + treated with anti-CD3, IL-1α, and IL-18 relative to IL-2 and IL-15. Mean of n = 6 skin donors. Statistical analysis was performed using a Friedman test followed by Dunn’s multiple comparisons test. Data in bar graphs (B) and (D) are shown as mean ± SD.

Article Snippet: Human skin–derived T lymphocytes were labeled with Cell Proliferation Dye eFluor 450 (10 μM, Thermo Fisher Scientific, catalog no. 65- 0842-85), washed with PBS, and subsequently stimulated for 6 days with rhuIL-2 (100 IU/ml; BioLegend, catalog no. 589108), rhuIL-15 (20 ng/ml; BioLegend, catalog no. 570603), in addition with mouse anti-human CD3 monoclonal antibody (1 μg/ml; Miltenyi Biotec, catalog no. 130-093-387, RRID:AB_1036144) alone or with rhuIL-1α (9 ng/ml; BioLegend, catalog no. 570004) and rhuIL-18 (9 ng/ml, BioLegend, catalog no. 592102) applied individually or in combination in cTexMacs medium.

Techniques: Produced, Derivative Assay, Ex Vivo, Expressing, Staining, Labeling, Cell Culture, Fluorescence, Flow Cytometry