Journal: Frontiers in Immunology

Article Title: Characterizing Foxp3 + and Foxp3 - T cells in the homeostatic state and after allo-activation: resting CD4 + Foxp3 + Tregs have molecular characteristics of activated T cells

doi: 10.3389/fimmu.2024.1292158

Figure Lengend Snippet: Antibodies and viability dyes used in flow cytometry.

Article Snippet: Live/dead , n/a , LIVE/DEADTM Fixable Near IR (780) Viability Kit (NIR) , eBioscience , L34994 , 1/1000.

Techniques: Cytometry, Staining

Journal: eBioMedicine

Article Title: Enhancing the efficacy of near-infrared photoimmunotherapy through intratumoural delivery of CD44–targeting antibody–photoabsorber conjugates

doi: 10.1016/j.ebiom.2025.105566

Figure Lengend Snippet: Intratumoural delivery of CD44-IR700 enhanced efficacy of photoimmunotherapy in vivo . (a) Schematic of experimental schedule. Mice bearing subcutaneous Lewis lung carcinoma tumours were treated with either no CD44-IR700 (Ctrl), CD44-IR700 via IV injection (CD44-IR700_IV), or CD44-IR700 via IT injection (CD44-IR700_IT), followed by NIR light irradiation on subsequent days. (b) The ratio of tumour-infiltrating CD8-positive T cells in live cells, as determined by flow cytometry on day 7 post-treatment, from the CD44-IR700_IV and CD44-IR700_IT groups (n = 6 per group). (c) Immunohistochemical staining of tumours 7 days after photoimmunotherapy. CD8-positive T cells were stained. The scale bar represents 50 μm. The numbers of CD8-positive T cells per view field were counted (n = 5 mice for each group). (d) Cytokine expression in tumours, as determined by quantitative PCR analysis on day 3 post-treatment with photoimmunotherapy, from the CD44-IR700_IV and CD44-IR700_IT groups (n = 5 per group). (e) Tumour volume monitoring. Tumour size was measured starting from the day of CD44-IR700 administration (day 0), and the change in size was expressed as a ratio relative to the initial volume on day 0 (n = 12 per group). (f) Tumour volume ratio on day 7. The graph shows the fold increase in tumour volume on day 7 compared with that on day 0 (n = 12 per group). (g) Representative images displaying tumours from each treatment group on day 7 post-administration. (h) Representative images of tumours excised on day 7 post-treatment from each therapeutic group. Data are presented as mean + standard error of the mean. Statistical difference was assessed using Mann–Whitney U tests, with significance denoted by asterisks (∗ p < 0.05 and ∗∗∗∗ p < 0.0001). CD44-IR700, IR700-conjugated anti-CD44 monoclonal antibody; CD, cluster of differentiation; NIR, near-infrared; Ctrl, control; IV, intravenous; IT, intratumoural; Ifnγ, interferon-γ; and Tnf, tumour necrosis factor-alpha.

Article Snippet: To assess the viability of LLC cells, 1 h after NIR irradiation, the cell counting kit-8 reagent (CK04, Dojindo) was added to each well and incubated at 37 °C for 1 h. The absorbance of the cell medium was measured at 450 nm using a Spectramax i3X microplate reader (Molecular Devices).

Techniques: In Vivo, IV Injection, Injection, Irradiation, Flow Cytometry, Immunohistochemical staining, Staining, Expressing, Real-time Polymerase Chain Reaction, MANN-WHITNEY, Control

Journal: eBioMedicine

Article Title: Enhancing the efficacy of near-infrared photoimmunotherapy through intratumoural delivery of CD44–targeting antibody–photoabsorber conjugates

doi: 10.1016/j.ebiom.2025.105566

Figure Lengend Snippet: The interval between intratumoural administration of CD44-IR700 and NIR irradiation, whether 30 min or 24 h, yields the same effect . (a) Schematic of experimental schedule. Lewis Lung carcinoma cell-induced subcutaneous tumours were established in mice. The mice were divided into three groups for the following treatments: CD44-IR700_IV_24h: IV administration of CD44-IR700 and IT of PBS, with NIR light irradiation 24 h after administration; CD44-IR700_IT_24h: IT administration of CD44-IR700 and IV injection of PBS, followed by NIR light irradiation 24 h later; and CD44-IR700_IT_30m: IT administration of CD44-IR700 and IV injection of PBS, with NIR light irradiation conducted 30 min post-administration. (b) Tumour volume measurement. The tumour volume was monitored from the day of CD44-IR700 administration (designated as day 0), and the change in tumour size was presented as a ratio to the volume on the day of administration (n = 14–18 per group). (c) Tumour volume ratio on day 7 relative to day 0 (n = 14–18 per group). Data are expressed as mean + standard error of the mean. Statistical difference was assessed using Kruskal–Wallis test, with asterisks denoting the levels of significance (∗ p < 0.05). ‘ns’ indicates a lack of statistical significance. CD44-IR700, IR700-conjugated anti-CD44 monoclonal antibody; PBS, phosphate-buffered saline; CD, cluster of differentiation; NIR, near-infrared; Ctrl, control; IV, intravenous; and IT, intratumoural.

Article Snippet: To assess the viability of LLC cells, 1 h after NIR irradiation, the cell counting kit-8 reagent (CK04, Dojindo) was added to each well and incubated at 37 °C for 1 h. The absorbance of the cell medium was measured at 450 nm using a Spectramax i3X microplate reader (Molecular Devices).

Techniques: Irradiation, IV Injection, Saline, Control