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 TCR/DC panel included TCRγδ Vio Blue (Miltenyi Biotec, cat. no.1 30-119-619), TCRαβ FITC (Miltenyi Biotec, cat. no. 130-113-538), CD83 PE (Miltenyi Biotec, cat. no. 130-110-503), CD3 Per CP Vio-770 (Miltenyi Biotec, cat. no. 130-113-141) and CD45 APC (Miltenyi Biotec, cat. no. 130-110-633).

Techniques: Enzyme-linked Immunospot, Whisker Assay

Journal: International Journal of Molecular Sciences

Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

doi: 10.3390/ijms27114875

Figure Lengend Snippet: Positive and negative control cells for both marker panels. ( A ) HeLa (white arrows): in the first panel positive for cytokeratin (CK), Vimentin (Vim), VEGF and in the second panel positive for p16INK4A, negative for PD-L1 and CD45. ( B ) T98G (red arrows): positive for Vim. ( C ) CaSki (yellow arrows): positive for CK, Vim, VEGF, PD-L1 and p16INK4A. ( D ) MCF-7 (green arrows): positive for CK. All cell lines were negative for CD45. Only hematopoietic cells showed CD45 positive staining. Scale bar = 10 μm. Images were processed using Zeiss ZEN 3.7 software and the figure was assembled using Microsoft PowerPoint.

Article Snippet: In the second round of immunofluorescence staining, anti-human PD-L1 conjugated with AF 647 antibody (Cat. No. ab209960, abcam, Cambridge, UK), anti-human p16INK4a conjugated with AF488 antibody (Cat. No. ab199756, abcam, Cambridge, UK), and, for counterstaining, anti-human CD45 antibody conjugated with PE (Cat. No. 130-110-770, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied, followed by image acquisition using the excitation filters mentioned above.

Techniques: Negative Control, Marker, Staining, Software

Journal: International Journal of Molecular Sciences

Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

doi: 10.3390/ijms27114875

Figure Lengend Snippet: Representative images of patient-derived DTCs with three different profiles. ( A ) DTC positive for Vim, VEGF and p16, negative for CK, PD-L1 and CD45. ( B ) CK and Vim positive while negative for VEGF, PD-L1, p16 and CD45. ( C ) DTC positive for Vim and PD-L1, negative for all other markers. Scale bar = 10 μm. Images were processed using Zeiss ZEN 3.7 software and figure was assembled using Microsoft PowerPoint.

Article Snippet: In the second round of immunofluorescence staining, anti-human PD-L1 conjugated with AF 647 antibody (Cat. No. ab209960, abcam, Cambridge, UK), anti-human p16INK4a conjugated with AF488 antibody (Cat. No. ab199756, abcam, Cambridge, UK), and, for counterstaining, anti-human CD45 antibody conjugated with PE (Cat. No. 130-110-770, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied, followed by image acquisition using the excitation filters mentioned above.

Techniques: Derivative Assay, Software

Journal: International Journal of Molecular Sciences

Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

doi: 10.3390/ijms27114875

Figure Lengend Snippet: Sequential multi-parameter immunofluorescent staining. After applying the first antibody panel (CK, Vim and VEGF), the slides were scanned and DTC profiles were assessed. Subsequently, releasable fluorochrome-conjugates were digested and the slides were subjected to the second antibody panel (PD-L1, p16, CD45) followed by scanning and DTC detection. Created with BioRender.com.

Article Snippet: In the second round of immunofluorescence staining, anti-human PD-L1 conjugated with AF 647 antibody (Cat. No. ab209960, abcam, Cambridge, UK), anti-human p16INK4a conjugated with AF488 antibody (Cat. No. ab199756, abcam, Cambridge, UK), and, for counterstaining, anti-human CD45 antibody conjugated with PE (Cat. No. 130-110-770, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied, followed by image acquisition using the excitation filters mentioned above.

Techniques: Staining

Journal: International Journal of Molecular Sciences

Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

doi: 10.3390/ijms27114875

Figure Lengend Snippet: CaSki cells before and after the release step. Representative immunofluorescence image of two CaSki cells spiked into bone marrow cells ( A ) before and ( B ) after the release step. The PE channel shows VEGF staining before staining with CD45, while nuclei were stained with DAPI (blue). Images were processed using Zeiss ZEN software and figure was assembled using Microsoft PowerPoint.

Article Snippet: In the second round of immunofluorescence staining, anti-human PD-L1 conjugated with AF 647 antibody (Cat. No. ab209960, abcam, Cambridge, UK), anti-human p16INK4a conjugated with AF488 antibody (Cat. No. ab199756, abcam, Cambridge, UK), and, for counterstaining, anti-human CD45 antibody conjugated with PE (Cat. No. 130-110-770, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied, followed by image acquisition using the excitation filters mentioned above.

Techniques: Immunofluorescence, Staining, Software