Journal: Cell Reports Medicine

Article Title: Potentiating immunotherapy in “immune-cold” solid tumors through orchestrating T cell immunity via tumor-specific genetic engineering

doi: 10.1016/j.xcrm.2025.102510

Figure Lengend Snippet: P αCD3&LIGHT improved anti-melanoma efficacy of CAR-T cells without obvious systemic toxicity (A) Therapeutic scheme of P αCD3&LIGHT in combination with CAR-T cells. hCD19-B16 melanoma-bearing mice were treated with four intravenous injections of P αCD3&LIGHT or other controls and two intravenous injections of CAR-T cells at different doses (0–4 × 10 6 cells per mouse). (B) Experimental timeline of blood serum collection for evaluating severe cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). (C) Flow cytometry analysis of CAR-T cells in tumor tissues of hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + CAR-T cells or other controls. n = 8. (D) Tumor weights of hCD19-B16 melanoma-bearing mice treated with 0–4 × 10 6 CAR-T cells. n = 9 or 10. (E–G) ELISA of SAA (E), IL-6 (F), and IL-1β (G) in blood serum of hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + CAR-T cells or other controls. n = 8. (H and I) Body temperature (H) and body weight (I) of hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + CAR-T cells or other controls. n = 10. (J) Radar map of fold changes of five CRS-related markers. The larger the area enclosed by the five markers, the more severe the CRS-related symptoms. (K) Representative immunohistochemistry images of CD11b-positive staining to indicate vascular leakage in lung, spleen, and liver tissues from hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + CAR-T cells or other controls. Scale bar: 100 μm. (L) Representative immunohistochemistry images of H&E staining to show the thickness of meninges in brain tissues from hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + CAR-T cells or other controls. Scale bar: 100 μm. (M) Therapeutic scheme of P αCD3&LIGHT in combination with 1.5 × 10 6 CAR-T cells. hCD19-B16 melanoma-bearing mice were treated with four intravenous injections of P αCD3&LIGHT or other controls and two intravenous injections of PBS or total 1.5 × 10 6 CAR-T cells. (N) Representative IVIS spectrum images of DiR-labeled CAR-T cells at tumor sites in hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells or other controls. (O–Q) Flow cytometry analysis of CD69-positive activated CAR-T cells (O), TCF-1-positive stem cell-like (P), and Ki67-positive self-renewing CAR-T cells (Q) in tumor tissues from hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells or other controls. n = 8. (R–T) ELISA of IFN-γ (R), TNF-α (S), and Gzm-B (T) in tumor tissues from hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells or other controls. n = 8. (U) Flow cytometry analysis of cleaved caspase-3-positive apoptotic hCD19-B16 tumor cells in tumor tissues from hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells or other controls. n = 8. (V–X) Individual (V) and average (W) tumor growth curves and survival curves (X) of hCD19-B16 melanoma-bearing mice after treatment with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells or other controls. n = 8–10. (Y) Magnetic resonance imaging (MRI) images of complete cured melanoma-bearing mice treated with P αCD3&LIGHT + 1.5 × 10 6 CAR-T cells. Data are represented as mean ± SD (error bars) from biological replicates. p values were determined by unpaired two-tailed Student’s t test for (C) and one-way ANOVA with Tukey’s test for (D), (O)–(U), and (W). n.s., not significant; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001. See also .

Article Snippet: Rabbit Anti-mouse CD11b Polyclonal Antibody , Servicebio , Cat# GB115689.

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Labeling, Magnetic Resonance Imaging, Two Tailed Test

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Longitudinal imaging evaluation of the inflammatory role of purinergic A 2 A receptors during subacute and chronic ischemic stroke

doi: 10.1177/0271678X251370835

Figure Lengend Snippet: Temporal ex vivo evaluation of A 2 AR expression after cerebral ischemia. Immunofluorescent labeling of A 2 ARs (blue), NeuN (orange), CD11b (red), and DAPI (gray) in the penumbra and core of the infarction in the control and day 3 groups is shown in four channels (a–c). The data show A 2 AR expression in NeuN + and CD11b + cells from the control group (day 0) and at days 1 and 3 after cerebral ischemia in both the ischemic penumbra and the core of infarction (d, e). Immunohistochemistry labeling of NeuN and CD11b in the whole brain section before (day 0) and at days 1 and 3 after ischemia (f, g). The protein expression of A 2 ARs during the first 3 days after ischemia was evaluated by immunoblotting (h, i) and autoradiography (j, k). ** p < 0.01 and *** p < 0.001 compared with day 0; & p < 0.05 and &&& p < 0.001 compared with day 1. The values are presented as scatter dot bar plots (means ± SD). Scale bars, 30 μm (top panels) and 10 μm (bottom panels).

Article Snippet: For the first set of sections ( n = 12), the primary antibodies used were: for A 2 ARs staining, rabbit anti-rat A 2 AR (1:50; Alomone Labs, Israel); for NeuN staining, guinea pig anti-rat NeuN (1:1000; Synaptic Systems, Goettingen, Germany); and for CD11b, mouse anti-rat CD11b (1:300; Serotec, Raleigh, NC, USA).

Techniques: Ex Vivo, Expressing, Labeling, Control, Immunohistochemistry, Western Blot, Autoradiography

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Longitudinal imaging evaluation of the inflammatory role of purinergic A 2 A receptors during subacute and chronic ischemic stroke

doi: 10.1177/0271678X251370835

Figure Lengend Snippet: Effect of A 2 ARs on neuroinflammation after cerebral ischemia. MRI (T 2 W; a) and PET axial images of [ 18 F]DPA-714 (b) in vehicle- and CGS-treated ischemic rats at the level of the ischemic lesion. The infarct volume was evaluated via MRI (T 2 W) at day 1 after ischemia (c), and the neurological score (d) was determined at day 1 (before the start of treatment) and day 7 after MCAO in vehicle ( n = 5) and CGS-treated ( n = 5) rats. In vivo [ 18 F]DPA-714 PET signals and ex vivo immunohistochemical evaluation of TSPO expression in CD11b-positive cells were quantified on day 7 after ischemia in both the striatum and cerebral cortex (e–j). *** p < 0.001 compared with day 1, * p < 0.05 compared with the vehicle. The values are presented as violin plots showing all points (means ± SD). Scale bars, 20 μm.

Article Snippet: For the first set of sections ( n = 12), the primary antibodies used were: for A 2 ARs staining, rabbit anti-rat A 2 AR (1:50; Alomone Labs, Israel); for NeuN staining, guinea pig anti-rat NeuN (1:1000; Synaptic Systems, Goettingen, Germany); and for CD11b, mouse anti-rat CD11b (1:300; Serotec, Raleigh, NC, USA).

Techniques: In Vivo, Ex Vivo, Immunohistochemical staining, Expressing

Journal: Frontiers in Oncology

Article Title: The development of pancreatic cancer is accompanied by significant changes in the immune response in genetically predisposed mice

doi: 10.3389/fonc.2025.1603293

Figure Lengend Snippet: Immune cells infiltration of pancreatic tissue with PC (Kras/Cre mice) and control (Cre mice) after Sal or CER injections, (A) representative images of CD20 + , CD3 + , CD4 + , CD8 + cells staining; scale bars represent 50 µm, (B) median number of CD20 + , CD3 + , CD4 + , and CD8 + cells, (C) representative images of staining CD11b + cells (scale bars represent 50 µm) and percentage of mice with CD11b + cells staining, scale bars represent 50 µm, n = 5 mice/group. Kras/Cre, mice with Kras mutation; Cre, mice without Kras mutation; Sal, saline; CER, cerulein; hpfs, high power fields. Statistically significant differences (p value < 0.05) between mice are marked * (for details see the text).

Article Snippet: In the next stage, the slides were incubated with rabbit anti-mouse antibodies CD11b (Biorbyt, Catalog Number: orb31554, dilution 1:100), CD20 (Biorbyt, Catalog Number: orb374711, dilution 1:200), CD3 (Biorbyt, Catalog Number: orb348965, dilution 1:100), CD4 (Biorbyt, Catalog Number: orb4830, dilution 1:100) and CD8 (Biorbyt, Catalog Number: orb323288, dilution 1:50) for 30 minutes at room temperature.

Techniques: Control, Staining, Mutagenesis, Saline

Journal: Frontiers in Oncology

Article Title: The development of pancreatic cancer is accompanied by significant changes in the immune response in genetically predisposed mice

doi: 10.3389/fonc.2025.1603293

Figure Lengend Snippet: Immune cells infiltration of pancreatic tissue with PC (Kras/Cre mice) and control (Cre ctr mice) after FMT and sham treatments, (A) representative images of staining CD20 + , CD3 + , CD4 + , CD8 + cells, scale bars represent 50 µm, (B) median number of CD20 + , CD3 + , CD4 + , CD8 + cells; (C) representative images of staining CD11b + cells (scale bars represent 50 µm) and percentage of mice with CD11b + cells staining estimeted as a weak (≤ 5 positive cells) or strong (>5 positive cells in site of view).; Kras/Cre, mice with Kras mutation; Cre, mice without Kras mutation; FMT, fecal microbiota transplantation; n = 5 mice/group; Statistically significant differences (p value < 0.05) between mice are marked * (for details see the text).

Article Snippet: In the next stage, the slides were incubated with rabbit anti-mouse antibodies CD11b (Biorbyt, Catalog Number: orb31554, dilution 1:100), CD20 (Biorbyt, Catalog Number: orb374711, dilution 1:200), CD3 (Biorbyt, Catalog Number: orb348965, dilution 1:100), CD4 (Biorbyt, Catalog Number: orb4830, dilution 1:100) and CD8 (Biorbyt, Catalog Number: orb323288, dilution 1:50) for 30 minutes at room temperature.

Techniques: Control, Staining, Mutagenesis, Transplantation Assay