Journal: Acta Pharmaceutica Sinica. B

Article Title: An immunostimulant nanomedicine enhances radioimmunotherapy by remodeling the tumor immunosuppressive landscape after radiotherapy

doi: 10.1016/j.apsb.2025.11.012

Figure Lengend Snippet: Immunosuppression after radiotherapy promotes breast cancer progression. Tumor growth curves (A) and weights (B) of mice before and after RT ( n = 6). (C) Survival curve of mice after different treatments ( n = 6). Tumor growth curves (D) and weights (E) of mice treated with RT and RT plus α -PD-L1 ( n = 6). (F) Survival curve of mice after different treatments ( n = 6). (G) Cluster analysis of differential expression genes between untreated and RT-treated tumors 48 h post-RT ( n = 3). (H) Gene Ontology (GO) analysis of tumor tissues before and after RT (select the top 10 for each item). (I) Heat map of differentially expressed genes related to apoptosis and immune suppression ( n = 3). (J) Immunofluorescence staining images of tumor tissue in saline and RT groups (scale bar = 20 μm). (K) Quantitative analysis of tumor-infiltrating CD45 + cells ( n = 6). Representative flow cytometry images (M) and quantitative analysis (L) of tumor-infiltrating MDSCs ( n = 6). (N) The TUNEL staining of tumor section (scale: 25 μm). (O) Adenosine content detection in tumor tissue ( n = 6). Data are presented as mean ± SD. ∗∗∗∗ P < 0.0001 determined by Student’s t-test.

Article Snippet: InVivo MAb anti-mouse PD-L1 was purchased from Bioxcell (USA).

Techniques: Quantitative Proteomics, Immunofluorescence, Staining, Saline, Flow Cytometry, TUNEL Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: An immunostimulant nanomedicine enhances radioimmunotherapy by remodeling the tumor immunosuppressive landscape after radiotherapy

doi: 10.1016/j.apsb.2025.11.012

Figure Lengend Snippet: FD@ATRA-enhanced radiotherapy combined with α -PD-L1 to enhance the efficacy of large-volume tumors. (A) Schematic diagram of the therapeutic process for evaluating the anti-tumor effects in a bilateral 4T1 tumor-bearing mouse model of large volume. (B) Distant tumor growth curves of mice after different treatments ( n = 5). (C) In vitro images of the distant tumors ( n = 5). (D) In vitro distant tumors mass ( n = 5). (E) Immunohistochemical staining of CD3 in tumor tissue slices after different treatments (scale bars = 100 μm). The images below were the corresponding enlarged parts (scale bars = 25 μm). (F) Immunohistochemical staining of CD8 in tumor tissue slices after various treatments (scale bars = 100 μm). The images below were the corresponding enlarged parts (scale bars = 25 μm). Data are presented as mean ± SD. ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001 determined by Student’s t -test.

Article Snippet: InVivo MAb anti-mouse PD-L1 was purchased from Bioxcell (USA).

Techniques: In Vitro, Immunohistochemical staining, Staining

Journal: NPJ Vaccines

Article Title: The co-delivery of adenovirus-based immune checkpoint vaccine elicits a potent anti-tumor effect in renal carcinoma

doi: 10.1038/s41541-023-00706-x

Figure Lengend Snippet: a , b The proliferation of lymphocytes isolated from the spleens of mice in each group after continuous stimulation of CAIX protein (10 μg/ml) was detected by the EdU assay. c Detection of T lymphocytes secreting IFN-γ by ELISPOT assay. d After stimulation with CAIX protein, the proportions of IL-2 + CD8 + T cells, IFN-γ + CD8 + T cells, and TNF-α + CD8 + T cells in splenocytes from each group were detected by flow cytometry. e – g Statistical analysis of the percentages of IL-2 + CD8 + T cells, TNF-α + CD8 + T cells, and IFN-γ + CD8 + T cells in d . Data presented as mean ± SD from one representative experiment of three performed. The different significance was set at *** p < 0.001 and **** p < 0.0001. Multiple groups of comparison data were analyzed by one-way ANOVA.

Article Snippet: For cell surface staining, cells were incubated with the following antibodies: PE-conjugated anti-human CAIX (R&D Systems, Cat. FAB2188P, 1:100), PE-conjugated anti-mouse CD3ε (BioLegend, Cat. 100308, 1:100), APC-conjugated anti-mouse PD-L1 (BioLegend, Cat. 124312, 1:100), APC-conjugated anti-mouse CD11c (BioLegend, Cat. 117310, 1:100), PerCP-Cy5.5-conjugated anti-mouse CD4 (BioLegend, Cat. 116012, 1:100), PerCP-Cy5.5-conjugated anti-mouse CD8α (BioLegend, Cat. 100734, 1:100), FITC anti-mouse CD49b (BioLegend, Cat. 108906, 1:100), PerCP-conjugated anti-mouse F4/80 (BioLegend, Cat. 123126, 1:100), FITC-conjugated anti-mouse CD11b (BioLegend, Cat. 101206, 1:100) for 1 h at 4 °C.

Techniques: Isolation, EdU Assay, Enzyme-linked Immunospot, Flow Cytometry, Comparison

Journal: NPJ Vaccines

Article Title: The co-delivery of adenovirus-based immune checkpoint vaccine elicits a potent anti-tumor effect in renal carcinoma

doi: 10.1038/s41541-023-00706-x

Figure Lengend Snippet: Ad-CAIX and Ad- PD-L1 vaccines were prepared and expressed in vitro. Three tumor models, including the subcutaneous, lung metastasis, and orthotropic tumor, were established, and intramuscular Ad vaccine immunization was performed. Ad-CAIX/Ad-PD-L1 could effectively enhance the induction and maturation of DCs and DC subsets, and promote strong tumor-specific CD8 + T cell immune responses. Ad-CAIX/Ad-PD-L1 vaccine could significantly inhibit tumor growth or lung metastasis in three models via DCs-mediated CD8 + T cell anti-tumor responses.

Article Snippet: For cell surface staining, cells were incubated with the following antibodies: PE-conjugated anti-human CAIX (R&D Systems, Cat. FAB2188P, 1:100), PE-conjugated anti-mouse CD3ε (BioLegend, Cat. 100308, 1:100), APC-conjugated anti-mouse PD-L1 (BioLegend, Cat. 124312, 1:100), APC-conjugated anti-mouse CD11c (BioLegend, Cat. 117310, 1:100), PerCP-Cy5.5-conjugated anti-mouse CD4 (BioLegend, Cat. 116012, 1:100), PerCP-Cy5.5-conjugated anti-mouse CD8α (BioLegend, Cat. 100734, 1:100), FITC anti-mouse CD49b (BioLegend, Cat. 108906, 1:100), PerCP-conjugated anti-mouse F4/80 (BioLegend, Cat. 123126, 1:100), FITC-conjugated anti-mouse CD11b (BioLegend, Cat. 101206, 1:100) for 1 h at 4 °C.

Techniques: Vaccines, In Vitro

Journal: Journal of Clinical Investigation

Article Title: Tumor endothelial marker 1–specific DNA vaccination targets tumor vasculature

doi: 10.1172/jci67382

Figure Lengend Snippet: Figure 4 Tem1-TT vaccination inhibits CT26 tumor vascularization. (A) Tem1-TT vaccination reduced tumor hemoglobin content. Tumors at approximately 200 mm3 were excised from TT- or Tem1-TT–vaccinated mice and inspected grossly. Tumors from Tem1-TT–vaccinated mice appeared pale relative to control tumors. Reduced hemoglobin levels in tumors from Tem1-TT–vaccinated mice were observed by ELISA. (B) Tem1-TT vaccine reduces tumor vascularity. Tumors from TT- or Tem1-TT–vaccinated mice were analyzed by Doppler ultrasound. Perfused tumor area and real blood flux are shown, measured and calculated by Doppler image analysis. (C) CT26 tumors from Tem1-TT–immunized mice had significantly decreased CD31 expression compared with TT vaccination. Also note the abnormal blood vessel shape. Original magnification, ×20. (D) CAIX expression was increased in tumors from Tem1-TT–immunized animals. CAIX expression was visualized by immunohistochemistry, and Caix was independently quantified by qRT-PCR in tumors from mice vaccinated with either TT or Tem1-TT. Original magnification, ×20. Data in A–D are mean ± SD of a representative experiment (n = 5 per group). Statistical analyses were performed with Student’s t test.

Article Snippet: Sections (6 μm thick) were stained for mouse CAIX (R&D Systems; catalog no. AF2344, DAB chromogen), CD31 (BD Biosciences — Pharmingen; clone 390, catalog no. 558737, DAB chromogen), and CD3 (BD Biosciences — Pharmingen; clone 15.5-2C11, catalog no. 550275, DAB chromogen) with hematoxylin as a counterstain.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemistry, Quantitative RT-PCR

Journal: Cancers

Article Title: Harnessing Induced Essentiality: Targeting Carbonic Anhydrase IX and Angiogenesis Reduces Lung Metastasis of Triple Negative Breast Cancer Xenografts

doi: 10.3390/cancers11071002

Figure Lengend Snippet: Sunitinib induces hypoxia and Carbonic Anhydrase IX (CAIX) expression in primary Triple Negative Breast Cancer (TNBC) tumors. ( a ) Images of MDA-MB-231 LM2-4Luc + primary tumor tissue sections harvested at increasing tumor volumes from animals administered either vehicle or 60 mg/kg sunitinib and immunohistochemically stained for the indicated markers. Scale bars: upper panels, 1 mm; lower panels, 200 μm. ( b to e ) Image-based quantification of ( b ) CD31 + blood vessels, ( c ) CAIX expression, ( d ) pimonidazole and ( e ) proliferation. Data show the mean ± standard error of the mean (SEM). n = 3 animals/group with 10 images/animal. * p < 0.05, *** p < 0.001.

Article Snippet: Formalin-fixed, paraffin-embedded (FFPE) tissues were sectioned (4–5 μm) and stained with primary antibodies against human CAIX (1:50, AF2188; R&D Systems, Minneapolis, MN, USA), CD31 (1:10, DIA-310; Histobiotec, Miami Beach, FL, USA), BrdU (1:10, Roche, Laval, QC, Canada) and pimonidazole (1:100, Hydroxyprobe, Burlington, MA, USA).

Techniques: Expressing, Staining

Journal: Cancers

Article Title: Harnessing Induced Essentiality: Targeting Carbonic Anhydrase IX and Angiogenesis Reduces Lung Metastasis of Triple Negative Breast Cancer Xenografts

doi: 10.3390/cancers11071002

Figure Lengend Snippet: SLC-0111 reduces vessel density and vascular permeability in primary tumors, and decreases lung and liver metastases. ( a ) Representative 3D maximum projections of whole mount primary tumor tissue slices showing CD31 + blood vessels (green) and fluorescently labeled dextran (red). Merged images demonstrate the presence of both intravascular (yellow) and extravasated (arrowheads) dextran. Scale bar = 100 μm. ( b ) Quantification of the number of vessels in whole mount primary tumor slices. Data show the mean ± standard error of the mean (SEM). n = 13–14 images/group. ** p < 0.01, *** p < 0.001. ( c ) Quantification of vascular permeability as assessed by relative area of extravasated dextran in whole mount primary tumor slices. Data show the mean ± SEM. n = 13–16 images/group. * p < 0.05, ** p < 0.01. ( d ) Representative 3D maximum projections of whole mount primary tumor tissue slices showing levels of CAIX expression (green) and the number of CD31 + blood vessels (red). Lower panels, merge. Scale bar = 100 μm. ( e ) Quantification of Carbonic Anhydrase IX (CAIX) expression in whole mount primary tumor tissue slices in panel d. Data show the mean ± SEM. n = 11–15 images/group. * p < 0.05, *** p < 0.001. ( f ) Representative images of lung and liver tissues from animals with MDA-MB-231 LM2-4Luc + orthotopic breast tumors and administered SLC-0111 and sunitinib, either alone or in combination. Visible metastatic nodules (arrows) are indicated. Scale bar = 1 cm. ( g – i ) Analysis of ( g ) lung weight ( n = 9/group), ( h ) liver weight as a function of whole animal body weight ( n = 9/group) and ( i ) number of metastatic nodules present on the liver surface ( n = 7–8/group). For each graph, data show the mean ± SEM. * p < 0.05.

Article Snippet: Formalin-fixed, paraffin-embedded (FFPE) tissues were sectioned (4–5 μm) and stained with primary antibodies against human CAIX (1:50, AF2188; R&D Systems, Minneapolis, MN, USA), CD31 (1:10, DIA-310; Histobiotec, Miami Beach, FL, USA), BrdU (1:10, Roche, Laval, QC, Canada) and pimonidazole (1:100, Hydroxyprobe, Burlington, MA, USA).

Techniques: Permeability, Labeling, Expressing

Journal: Cancers

Article Title: Harnessing Induced Essentiality: Targeting Carbonic Anhydrase IX and Angiogenesis Reduces Lung Metastasis of Triple Negative Breast Cancer Xenografts

doi: 10.3390/cancers11071002

Figure Lengend Snippet: Administration of SLC-0111 reduces lung metastases. ( a ) Representative 3D maximum projections of whole mount lung tissue slices from tumor-bearing mice, showing vimentin-positive metastases (cyan) and CD31 + blood vessels (red). Scale bar = 100 μm. ( b ) Analysis of vimentin positive metastases in whole mount lung tissue sections described in panel a. Data show the mean ± standard error of the mean (SEM). n = 8–10 images/group. * p < 0.05, *** p < 0.001. ( c ) Representative 3D maximum projections of whole mount lung tissue slices from tumor-bearing mice, showing Carbonic Anhydrase IX (CAIX)-positive metastases (green) and CD31 + blood vessels (red). Bar = 100 μm. ( d ) Analysis of CAIX positive metastases in the whole mount lung tissue sections described in panel c. Data show the mean ± SEM. n = 5–8 images/group: No statistically significant differences were observed among the groups.

Article Snippet: Formalin-fixed, paraffin-embedded (FFPE) tissues were sectioned (4–5 μm) and stained with primary antibodies against human CAIX (1:50, AF2188; R&D Systems, Minneapolis, MN, USA), CD31 (1:10, DIA-310; Histobiotec, Miami Beach, FL, USA), BrdU (1:10, Roche, Laval, QC, Canada) and pimonidazole (1:100, Hydroxyprobe, Burlington, MA, USA).

Techniques:

Journal: Cancers

Article Title: Harnessing Induced Essentiality: Targeting Carbonic Anhydrase IX and Angiogenesis Reduces Lung Metastasis of Triple Negative Breast Cancer Xenografts

doi: 10.3390/cancers11071002

Figure Lengend Snippet: Model for targeting angiogenesis and Carbonic Anhydrase IX (CAIX) to reduce metastasis of Triple Negative Breast Cancer (TNBC). Exposure of tumors to anti-angiogenic agents such as sunitinib leads to reduced tumor growth. However, major consequences can include enhanced hypoxia and increased vascular permeability. The exacerbation of hypoxia results in the upregulation of CAIX expression by tumor cells and the potentiation of metastasis. Inhibition of CAIX activity inhibits metastasis through several mechanisms, as shown by previous studies, but may also play a role in reducing vascular permeability and contributing to vascular normalization, potentially reducing the invasion of tumor cells to distant sites.

Article Snippet: Formalin-fixed, paraffin-embedded (FFPE) tissues were sectioned (4–5 μm) and stained with primary antibodies against human CAIX (1:50, AF2188; R&D Systems, Minneapolis, MN, USA), CD31 (1:10, DIA-310; Histobiotec, Miami Beach, FL, USA), BrdU (1:10, Roche, Laval, QC, Canada) and pimonidazole (1:100, Hydroxyprobe, Burlington, MA, USA).

Techniques: Permeability, Expressing, Inhibition, Activity Assay

Journal: Journal of Translational Medicine

Article Title: Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer?

doi: 10.1186/1479-5876-11-214

Figure Lengend Snippet: Summary of the immunocytochemical analysis of CNHCs with antibodies against CD45, CD31, and CAIX

Article Snippet: The filters were incubated with primary antibodies directed against CAIX (rabbit IgG, NB100-417, Novus Biologicals, Littleton, USA; 3.5 μg/ml ), CD31 (mouse IgG1, M0823, Dako, Glostrup, Denmark; 1 μg/ml) or CD45 (mouse IgG1, M0855, Dako, Glostrup, Denmark; 2.4 μg/ml) diluted in Antibody Diluent (Dako, Glostrup, Denmark) for 30 min at RT followed by application of primary antibody enhancer (Dako, Glostrup, Denmark) for 15 min.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer?

doi: 10.1186/1479-5876-11-214

Figure Lengend Snippet: Immunocytochemical analysis of CNHCs with antibodies against the RCC marker CAIX. Clusters of CNHCs cytomorphologically classified as uncertain malignant (−UMF) with cytoplasmic positive staining with antibodies against the RCC marker CAIX (A) . Clusters of CNHC-UMF and -BF without reactivity for CAIX antibodies ( B and C , respectively). A single CNHC-MF with positive cytoplasmic (D) and without staining for CAIX (E) . Single CAIX-negative CNHC-UMF and -BF ( F and G , respectively).

Article Snippet: The filters were incubated with primary antibodies directed against CAIX (rabbit IgG, NB100-417, Novus Biologicals, Littleton, USA; 3.5 μg/ml ), CD31 (mouse IgG1, M0823, Dako, Glostrup, Denmark; 1 μg/ml) or CD45 (mouse IgG1, M0855, Dako, Glostrup, Denmark; 2.4 μg/ml) diluted in Antibody Diluent (Dako, Glostrup, Denmark) for 30 min at RT followed by application of primary antibody enhancer (Dako, Glostrup, Denmark) for 15 min.

Techniques: Marker, Staining