Journal: Journal for Immunotherapy of Cancer

Article Title: SP140 inhibits STAT1 signaling, induces IFN-γ in tumor-associated macrophages, and is a predictive biomarker of immunotherapy response

doi: 10.1136/jitc-2022-005088

Figure Lengend Snippet: Overexpression of SP140 induces the production of inflammatory cytokines and chemokines and reprogram macrophages to induce TAM-mediated tumor cytotoxicity. (A, B) Control CRISPR/dCAS9 (control) or SP140 CRISPR/dCAS9 (SP140 overexpression) was transfected in naïve macrophages, and the expression of CD80 and CD86 was quantified by flow cytometry. (C–E) SP140 CRISPR/dCAS9 (SP140 OE) or scramble control (control CRISPR/dCAS9) were introduced to the naïve macrophages, and a multiplex fluorescence BioLegend assay was used to identify the levels of CXCL10, IFN-γ, and IL-12 (P70) in the supernatant after 48 hours. LPS (100 nM) was administered after 24 hours. (F–I) Correlation of SP140 expression and IFNG, CXCL10, IL-12B, and IL-12A levels in over 500 HNSCCs in the TCGA dataset. (J) Control CRISPR/dCAS9 or SP140 CRISPR/dCAS9 was introduced in TAMs isolated from syngeneic HNSCC tumor and cocultured with murine SCC7 cells. Cell viability after 48 hours of coculture was quantified with a quantitative viability assay kit. Data are presented based on the fold change of non-treated control. *P<0.05, ***P<0.001, ****P<0.0001. HNSCC, head and neck squamous cell carcinoma; IFN-γ, interferon gamma; IL, interleukin; SP140, speckled protein 140; TAM, tumor-associated macrophage; TCGA, The Cancer Genome Atlas.

Article Snippet: A flow cytometry panel consisting of human CD80 (PE, BioLegend), human CD86 (BV711, BioLegend), and human CD206 (PerCP-Cy5.5, BioLegend) was used for the characterization of macrophages.

Techniques: Over Expression, Control, CRISPR, Transfection, Expressing, Flow Cytometry, Multiplex Assay, Fluorescence, Isolation, Viability Assay

Journal: Central European Journal of Immunology

Article Title: Macrophage M2 polarization induced by ANKRD22 in lung adenocarcinoma facilitates tumor angiogenesis

doi: 10.5114/ceji.2025.149372

Figure Lengend Snippet: Fig. 2. Cont. F-H) Flow cytometry was used to detect the effects of ANKRD22 overexpression or knockdown on the proportions of CD68+CD86+ and CD68+CD206+ macrophages. All experiments were conducted with 3 independent replicates. The data are presented as mean ± standard deviation. *p < 0.05

Article Snippet: The cells were incubated with PerCP Anti-Human CD68 Antibody (1 : 100; 333813, BioLegend, USA), PE Anti-Human CD86 Antibody (1 : 100; E-AB-F1012D, Elabscience, China), and/or FITC AntiHuman CD206/MMR Antibody (1 : 100; E-AB-F1161C, Elabscience, China) for 15 min. After washing with PBS, the cells were resuspended, and the flow cytometry data were collected and analyzed using Agilent’s flow analysis software.

Techniques: Flow Cytometry, Over Expression, Knockdown, Standard Deviation

Journal: Central European Journal of Immunology

Article Title: Macrophage M2 polarization induced by ANKRD22 in lung adenocarcinoma facilitates tumor angiogenesis

doi: 10.5114/ceji.2025.149372

Figure Lengend Snippet: Fig. 2. Cont. I) Flow cytometry was used to detect the ef- fects of ANKRD22 overexpression or knockdown on the pro- portions of CD68+CD86+ and CD68+CD206+ macrophages. J) WB was used to detect the effects of ANKRD22 over- expression or knockdown on the protein expression levels of CD80 and CD163. All experiments were conducted with 3 independent replicates. The data are presented as mean ± standard deviation. *p < 0.05

Article Snippet: The cells were incubated with PerCP Anti-Human CD68 Antibody (1 : 100; 333813, BioLegend, USA), PE Anti-Human CD86 Antibody (1 : 100; E-AB-F1012D, Elabscience, China), and/or FITC AntiHuman CD206/MMR Antibody (1 : 100; E-AB-F1161C, Elabscience, China) for 15 min. After washing with PBS, the cells were resuspended, and the flow cytometry data were collected and analyzed using Agilent’s flow analysis software.

Techniques: Flow Cytometry, Over Expression, Knockdown, Expressing, Standard Deviation

Journal: Cancer immunology, immunotherapy : CII

Article Title: CD33 BiTE ® molecule-mediated immune synapse formation and subsequent T-cell activation is determined by the expression profile of activating and inhibitory checkpoint molecules on AML cells.

doi: 10.1007/s00262-023-03439-x

Figure Lengend Snippet: Fig. 1 T-cell co-signaling receptors are expressed on leukemic cells and transduced Ba/F3 sublines. A MFI ratio of cell surface expression of CD80 (green), CD86 (blue) and PD-L1 (orange) in CD45dimSSClow primary AML samples (n = 107–377). MFI ratios ≥ 1.5 (dashed lines) indicate positivity. B Percentage of CD80-, CD86- and PD-L1-posi-

Article Snippet: Human cDNA encoding CD80, CD86 and PD-L1 was purchased from R&D Systems (RDC1086, RDC0693 and RDC1087, respectively) and subcloned into the retroviral expression vector MSCV-neo.

Techniques: Expressing

Journal: Cancer immunology, immunotherapy : CII

Article Title: CD33 BiTE ® molecule-mediated immune synapse formation and subsequent T-cell activation is determined by the expression profile of activating and inhibitory checkpoint molecules on AML cells.

doi: 10.1007/s00262-023-03439-x

Figure Lengend Snippet: Fig. 2 AMG 330 induces TCR triggering characterized by CD45 exclusion from and CD33 clustering within the synapse. A Repre- sentative spinning disc confocal microscope images of AMG 330 (BiTE® molecule) and c BiTE molecule-mediated conjugates formed of a CD33-transduced Raji B cell and a reconstituted HEK-T cell. B Line profiles of CD45 (green), CD33 (blue), and AMG 330 (red) intensities across a conjugate interface equivalent to that shown in a representative image in panel A. C Total number of AMG 330-induced T-cell–CD33+ CD86± PD-L1± Ba/F3 cell con- jugates after 20 min, assessed by flow cytometry. D Representative

Article Snippet: Human cDNA encoding CD80, CD86 and PD-L1 was purchased from R&D Systems (RDC1086, RDC0693 and RDC1087, respectively) and subcloned into the retroviral expression vector MSCV-neo.

Techniques: Microscopy, Flow Cytometry

Journal: Cancer immunology, immunotherapy : CII

Article Title: CD33 BiTE ® molecule-mediated immune synapse formation and subsequent T-cell activation is determined by the expression profile of activating and inhibitory checkpoint molecules on AML cells.

doi: 10.1007/s00262-023-03439-x

Figure Lengend Snippet: Fig. 3 Checkpoint molecule expression on target cells modu- lates AMG 330-mediated cytotoxicity and T-cell function. A AMG 330-mediated cytolytic capacity, B proliferation, C gran- zyme B expression and cytokine secretion of HD T-cell subsets after co-culture with CD33+ CD86± PD-L1± Ba/F3 sublines for 72 h. D AMG 330-mediated cytolytic capacity in BM and spleen and E CD69

Article Snippet: Human cDNA encoding CD80, CD86 and PD-L1 was purchased from R&D Systems (RDC1086, RDC0693 and RDC1087, respectively) and subcloned into the retroviral expression vector MSCV-neo.

Techniques: Expressing, Cell Function Assay, Co-Culture Assay

Journal: Materials Today Bio

Article Title: Microalgal-enhanced cerium oxide nanotherapeutics for alleviating inflammatory bowel disease via scavenging reactive oxygen species and modulating gut microbiota in colitis

doi: 10.1016/j.mtbio.2025.101945

Figure Lengend Snippet: The protective effect of SP@CSC on HT29 cells under oxidative stress conditions. Representative images of intracellular ROS in HT29 cells under different treatments, as indicated by CLSM (a) and flow cytometry analysis (b), following stimulation with H 2 O 2 for 24 h. (c) Representative fluorescent images of JC-1 staining showing the mitochondrial membrane potential of H 2 O 2 -stimulated HT29 cells after treated with different formulations. JC-1 aggregates (red) and monomer (green) (d) Semi-quantitative analysis of the ratio of JC-1 aggregates (red) and monomer (green). (e) Annexin V-FITC/PI-PE staining comparing the differences in HT29 apoptosis among different groups. (f) Quantitative results of Annexin V+ cell rate. Data was shown as mean ± standard deviation (S.D) ( n = 3 per group). Statistical significance was performed by one-way ANOVA test with a Tukey's post hoc test. ∗ P < 0.05, ∗∗ P < 0.01. Data are presented as mean ± S.D., with n = 3 per group. Statistical significance was determined using one-way ANOVA followed by Tukey's post hoc test. ∗ P < 0.05, ∗∗ P < 0.01. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Antibodies for flow cytometry, specifically CD206-PE and CD86-FITC, along with ELISA kits for catalase (CAT), superoxide dismutase (SOD), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-17 (IL-17), were purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Flow Cytometry, Staining, Membrane, Standard Deviation

Journal: Materials Today Bio

Article Title: Microalgal-enhanced cerium oxide nanotherapeutics for alleviating inflammatory bowel disease via scavenging reactive oxygen species and modulating gut microbiota in colitis

doi: 10.1016/j.mtbio.2025.101945

Figure Lengend Snippet: Characterization of CSC NGs and SP@CSC. (a) Photographic of CeO 2 , CS NGs and CSC NGs. (b) TEM image of CeO 2 , CS, CSC NGs. (c) The hydrodynamic size distribution of CS, CeO 2 , and CSC NGs. (d) UV spectra of the indicated materials. XPS survey spectrum of Ce 3d in CeO 2 (e) and CSC NGs (f), and the semi-quantitation of Ce 3+ /Ce 4+ . (g) Cerium content of SP@CSC under different weight ratios of SP and CeO 2 . (h) Zeta-potential of CeO 2 , CSC NGs, SP, and SP@CSC (n = 3). (i) SEM images of SP and SP@CSC. (j) CLSM imaging of SP@CSC. The SP exhibited autofluorescence (Cy5 channel) and CSC was labeled by FITC. (k) Releasing curves of CeO 2 from SP@CSC in SGF and SIF. Data are presented as mean ± standard deviation (S.D.), with 3 replicates per group. Statistical significance was determined using one-way ANOVA followed by Tukey's post hoc test. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Antibodies for flow cytometry, specifically CD206-PE and CD86-FITC, along with ELISA kits for catalase (CAT), superoxide dismutase (SOD), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-17 (IL-17), were purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Quantitation Assay, Zeta Potential Analyzer, Imaging, Labeling, Standard Deviation

Journal: Materials Today Bio

Article Title: Microalgal-enhanced cerium oxide nanotherapeutics for alleviating inflammatory bowel disease via scavenging reactive oxygen species and modulating gut microbiota in colitis

doi: 10.1016/j.mtbio.2025.101945

Figure Lengend Snippet: SP@CSC alleviate IBD Inducing Factors in Macrophages. Cytoprotective effect of different treatments against H 2 O 2 -induced oxidative stress in RAW264.7 cells (a) and HT-29 cells (b). (c) Flow cytometry analysis of DCFH-DA staining in LPS-induced (1 μg/mL) RAW264.7 cells under various treatment conditions, along with (d) the corresponding quantitative evaluation. (e) Flow cytometry analysis of the proportions of CD86-positive and CD206-positive macrophages after LPS stimulation for 24 h. (f) Quantification of CD86-positive and CD206-negative cells across all groups. ELISA assays of typical proinflammatory of TNF-α (g), IL-6 (h). (i) Western blot assay for Nrf2, and HO-1expression of Raw 264.7 cells after LPS stimulation and treated with different treatments. Data are presented as mean ± S.D (n = 3 per group). Statistical significance was assessed using one-way ANOVA test. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Antibodies for flow cytometry, specifically CD206-PE and CD86-FITC, along with ELISA kits for catalase (CAT), superoxide dismutase (SOD), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-17 (IL-17), were purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

Techniques: Flow Cytometry, Staining, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Materials Today Bio

Article Title: Microalgal-enhanced cerium oxide nanotherapeutics for alleviating inflammatory bowel disease via scavenging reactive oxygen species and modulating gut microbiota in colitis

doi: 10.1016/j.mtbio.2025.101945

Figure Lengend Snippet: Therapeutic effects and immunomodulation properties of SP@CSC in vivo. (a) Overall design of animal experiments. Balb/c mice were fed with 3 % DSS for 7 consecutive days and orally administered different treatments or PBS every other day, starting from day 2, for a total of seven administrations. (b) Daily body weight changes in mice over 9 days (n = 5). (c) Colon length measurements for each group (n = 5). (d) DAI score for each group on day 9 (n = 5). (e) Representative photographs and hematoxylin and eosin-stained images of the retrieved colon tissues. Representative immunofluorescence images of F4/80 (red) (f) and CD86 (green) (g) in colonic tissues. (h) Semi-quantification of F4/80-positive areas (n = 3). (i) Semi-quantification of CD86-positive areas (n = 3). The levels of IL-6 (j), TNF-α (k), IL-17 (l) in the colon determined by ELISA assay (n = 3). Data are presented as mean ± S.D. ∗ P < 0.05 and ∗∗ P < 0.01 indicate statistical significance, as determined by one-way ANOVA followed by Tukey's post hoc test. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Antibodies for flow cytometry, specifically CD206-PE and CD86-FITC, along with ELISA kits for catalase (CAT), superoxide dismutase (SOD), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-17 (IL-17), were purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

Techniques: In Vivo, Staining, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Materials Today Bio

Article Title: Microalgal-enhanced cerium oxide nanotherapeutics for alleviating inflammatory bowel disease via scavenging reactive oxygen species and modulating gut microbiota in colitis

doi: 10.1016/j.mtbio.2025.101945

Figure Lengend Snippet: In vivo distribution of SP@CSC. (a) Time-dependent in vivo fluorescence images of Balb/c nude mice following intragastric administration of IR783-labeled CSC NGs and SP@CSC (n = 3). (b) Quantification of fluorescence intensity in the mice. (c) Ex vivo fluorescence images of major organs, including heart, liver, spleen, lung, kidney, and gastrointestinal (GI) tract, collected at different time points after intragastric administration, along with (d) quantification of fluorescence intensity in these organs. (e) Fluorescence images of colon tissues from mice gavaged with FITC-labeled CSC NGs and SP@CSC. (f) Schematic illustration of SP@CSC delivery.

Article Snippet: Antibodies for flow cytometry, specifically CD206-PE and CD86-FITC, along with ELISA kits for catalase (CAT), superoxide dismutase (SOD), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-17 (IL-17), were purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China).

Techniques: In Vivo, Fluorescence, Labeling, Ex Vivo

Journal: Cancers

Article Title: Isolation of Two Novel Human Anti-CTLA-4 mAbs with Intriguing Biological Properties on Tumor and NK Cells

doi: 10.3390/cancers12082204

Figure Lengend Snippet: Binding of the novel anti-CTLA-4 mAbs to CD28-Fc and to cynomologous CTLA-4-Fc. Cross-reactivity of ID-1 and ID-8 mAbs to CD28-Fc ( A ) or to monkey CTLA-4-Fc proteins ( B ). Ipilimumab was used in parallel assays as a control.

Article Snippet: The following recombinant proteins were used: Recombinant Human CTLA-4 Fc Chimera; Recombinant Human B7-1/CD80 Fc Chimera Protein; Recombinant Human B7-2/CD86 Fc Chimera Protein; Recombinant Human CD28 Fc protein; Recombinant Cynomolgus Monkey CTLA-4 Fc Chimera Protein; Recombinant Human IgG1 Fc Protein (all from R & D Systems, Minneapolis, MN, USA); Human CTLA-4 Protein (His & Fc Tag); CTLA-4 Protein, Mouse, Recombinant (Fc Tag) (both from Sino Biological, Wayne, PA, USA).

Techniques: Binding Assay, Control