Journal: NPJ Vaccines

Article Title: A KIF20A-based thermosensitive hydrogel vaccine effectively potentiates immune checkpoint blockade therapy for hepatocellular carcinoma

doi: 10.1038/s41541-024-01060-2

Figure Lengend Snippet: a CD80 and CD86 levels of BMDCs in flow cytometry (The gating strategy was shown in the Supplementary Fig. ). b Quantification of the proportion of CD80 + CD86 + BMDCs. c MHC-I level of BMDCs indicated by flow cytometry (The gating strategy was shown in the Supplementary Fig. ). d Quantification of the proportion of MHC-I + BMDCs. e MHC-II level of BMDCs in flow cytometry (The gating strategy was shown in the Supplementary Fig. ). f Quantification of MHC-II + BMDCs. g CFDA-SE level in CD3 + T cells (The gating strategy was shown in the Supplementary Fig. ). h Quantification of T cell proliferation rate. i CLSM images of infiltrated CD8a + T cells into Hepa1-6 cell spheroids. j Quantification of T cell infiltration.

Article Snippet: Cells were subsequently stained with APC-conjugated anti-mouse CD11c, FITC-conjugated anti-mouse CD80, and PE-conjugated anti-mouse CD86 antibodies (Elabscience, China) for flow cytometry analysis (Beckman, USA).

Techniques: Flow Cytometry

Journal: NPJ Vaccines

Article Title: A KIF20A-based thermosensitive hydrogel vaccine effectively potentiates immune checkpoint blockade therapy for hepatocellular carcinoma

doi: 10.1038/s41541-024-01060-2

Figure Lengend Snippet: a Scheme illustration of in vivo treatment and monitoring (Created by the authors with Photoshop v22.5.6). b Bioluminescence images of tumor-bearing mice in different groups. c Relative signal intensity of tumors in different groups. d Body weight of the mice in different groups. e Flow cytometry analyzing CD80 and CD86 in CD11c + dendritic cells in mouse inguinal lymph nodes (Gating strategy was shown in the Supplementary Fig. ). f Quantification of the percentage of CD80 + CD86 + cells in CD11c + dendritic cells. g Flow cytometry analyzing CD8a and CD4 of T cells (CD45 + CD3 + ) in tumor tissues (Gating strategy was shown in the Supplementary Fig. ). h Quantification of the percentage of CD8a + cells in T cells.

Article Snippet: Cells were subsequently stained with APC-conjugated anti-mouse CD11c, FITC-conjugated anti-mouse CD80, and PE-conjugated anti-mouse CD86 antibodies (Elabscience, China) for flow cytometry analysis (Beckman, USA).

Techniques: In Vivo, Flow Cytometry

Journal:

Article Title: Lack of Tumor Necrosis Factor Alpha Induces Impaired Proliferation of Hepatitis B Virus-Specific Cytotoxic T Lymphocytes

doi: 10.1128/JVI.77.4.2469-2476.2003

Figure Lengend Snippet: Comparison of DC function in TNF-α+/+ and TNF-α−/− mice. CD11c+ cell separation was carried out by using the VarioMACS system. (A) Flow cytometric analysis of the expression of the MHC-I molecule H-2Ld (restriction element for HBsAg-specific CTL), the MHC-II molecule I-A/I-E, CD80, and CD86 in TNF-α+/+ and TNF-α−/− DCs. All samples were assayed in duplicate. (B) The allostimulatory capacity of CD11c+ DCs from TNF-α+/+ and TNF-α−/− mice was assessed by the MLR as described in Materials and Methods with allogeneic murine CD4+ T cells from AKR mice (H-2k). Each data point and error bar represent the mean and SD, respectively, of results for triplicate samples.

Article Snippet: To evaluate the expression intensities of MHC and costimulatory molecules, mouse or rat biotin-conjugated anti-mouse MHC-I ( H - 2L d ), MHC-II ( I - A d / I - E d ), PE-labeled streptavidin (BD PharMingen), rat PE-conjugated anti-mouse CD80 and CD86 (Cedarlane, Ontario, Canada) were used for staining.

Techniques: Expressing

Journal: Stem cell research & therapy

Article Title: Migrasomes derived from human umbilical cord mesenchymal stem cells: a new therapeutic agent for ovalbumin-induced asthma in mice.

doi: 10.1186/s13287-025-04145-4

Figure Lengend Snippet: Fig. 3 Administration of hUCMSC-migrasomes diminished Th2 response in allergic asthma by suppressing DC maturation. A The expression of IL-4, IL-5 and IL-13 in the lung of mice in each group were detected by real-time PCR (n = 6). B Gating strategy and representative flow cytometric images for the analysis of CD45+CD4+CCR4+CXCR3− Th2 cells in the lungs of mice from each group. C Graphs showing the % of CD45+CD4+CCR4+CXCR3−Th2 cells in the lungs of mice from each group (n = 6). D Gating strategy and representative flow cytometric dot plots (left) for the analysis of CD11b+ DCs in the lungs of allergic mice and the numbers of CD11b+ DCs (right) in the lungs of mice from each group (n = 6). E The mean MFI of CD80, CD86 and MHC-II on CD11b+ DCs (n = 4–6). F The expression of IL-6 in the lung of mice were detected by real-time PCR (n = 6). G Distribution of Dir-labeled migrasomes in OVA-induced mice lung after tail vein administration (17-day injection and 18-day detected). H Representative immunofluorescence images of Dil-labeled migrasomes (red) taken up by CD11c+ cells or CD4+ cells. Data were presented as mean ± SD. A one-way analysis of variance (Tukey Kramer post hoc tests) was per formed on the data. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The cell pellets were then stained with a range of monoclonal antibodies including FITC anti-mouse CD45 (eBioscience, San Diego, CA), eflour 450 anti-mouse CD4 (eBioscience), PE- anti-mouse CCR4 (Biolegend), APC anti-mouse CXCR3 (Biolegend), eflour 450 anti-mouse CD11c (eBioscience), FITC anti-mouse CD11b (Elabscience, Wuhan, China), PE/Cyanine7 anti-mouse CD80 (Elabscience), PE anti-mouse CD86 (eBioscience) and APC anti-mouse MHC-II (eBioscience).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Labeling, Injection, Immunofluorescence

Journal: Stem cell research & therapy

Article Title: Migrasomes derived from human umbilical cord mesenchymal stem cells: a new therapeutic agent for ovalbumin-induced asthma in mice.

doi: 10.1186/s13287-025-04145-4

Figure Lengend Snippet: Fig. 4 HUCMSC-migrasomes impaired DC maturation and function in vitro. A BMDCs incubated with Dil-labeled migrasomes under fluorescence micro scope (white bar = 25 μm). B The cell viability of BMDCs treated by hUCMSC-migrasomes for 24 h (left) and 48 h (right) (n = 4). C BMDCs were isolated from wild-type BALB/c mice and cultured at a density of 2 × 10⁵ cells/mL. The cells were stimulated with OVA (100 µg/mL) and LPS (10 ng/mL) for 48 h, with or without hUCMSC-migrasomes (20 µg/mL) pre-treated. After that, cells were collected for the detection of DC maturation markers including CD80, CD86 and MHC-II (n = 3). The representative images for the detection of DC maturation markers are displayed. D Statistical analysis of the mean MFIs of CD80, CD86 and MHC-II of BMDCs (n = 3). E Statistical analysis of the expression of IL-6 in BMDCs by real-time PCR (n = 3). F BMDCs (2 × 104) from WT mice were pre-treated with or without migrasomes and then pulsed with OVA323–339 and LPS and incubated at a 1:5 ratio with splenic CD4+ T cells isolated from OT-II mice (1 × 105) for 72 h. G Statistical analysis of the expression of IL-4, IL-5, IL-13 in collected cell by real-time PCR (n = 3). Data were presented as mean ± SD. A one-way analysis of variance (Tukey Kramer post hoc tests) was performed on the data. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The cell pellets were then stained with a range of monoclonal antibodies including FITC anti-mouse CD45 (eBioscience, San Diego, CA), eflour 450 anti-mouse CD4 (eBioscience), PE- anti-mouse CCR4 (Biolegend), APC anti-mouse CXCR3 (Biolegend), eflour 450 anti-mouse CD11c (eBioscience), FITC anti-mouse CD11b (Elabscience, Wuhan, China), PE/Cyanine7 anti-mouse CD80 (Elabscience), PE anti-mouse CD86 (eBioscience) and APC anti-mouse MHC-II (eBioscience).

Techniques: In Vitro, Incubation, Labeling, Fluorescence, Isolation, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

Journal: Arthritis Research & Therapy

Article Title: Interferon-induced protein IFIT4 is associated with systemic lupus erythematosus and promotes differentiation of monocytes into dendritic cell-like cells

doi: 10.1186/ar2475

Figure Lengend Snippet: Comparison of the phenotypic profiles of DCLCs primed with or without IFIT4 over-expression. (a) Surface antigens of normal THP-1 cells were examined by flow cytometry. (b) To analyze the effect of IFIT4 on phenotypic changes of dendritic cell-like cells (DCLCs) during the process of differentiation, THP-1 cells were transfected with pEGFP-IFIT4 or pEGFP-C1; 36 hours later, cells were stimulated with granulocyte-macrophage colony-stimulating factor (GM-CSF; 50 ng/ml) and IL-4 (20 ng/ml) for 90 hours to generate DCLCs. These DCLCs primed with pEGFP-IFIT4 or pEGFP-C1 transfection were incubated with fluorochrome-conjugated monoclonal antibodies (mAbs) and the antigens of CD40, CD80, CD86, CD83, HLA-DR, CD14 and CD1a on the surface of those DCLCs were analyzed by flow cytometry. Appropriate fluorochrome or isotype control mAbs of each antibody species were used as negative controls. Shaded histograms represent isotype control antibodies. The thick line represents DCLCs primed with pEGFP-IFIT4 transfection, whereas the slender lines represent DCLCs primed with pEGFP-C1 transfection. All experiments were performed three times and a set of representative histograms was presented. IFIT4, interferon-induced protein with tetratricopeptide repeats 4.

Article Snippet: FITC anti-human CD40 (Catalogue no: FAB617F) and PE anti-human CD80 (Catalogue no: FAB140P) were from R&D Systems, Minneapolis, MN, USA.

Techniques: Comparison, Over Expression, Flow Cytometry, Transfection, Incubation, Bioprocessing, Control

Journal: Life sciences

Article Title: TPX2 influences the regulation of macrophage polarization via the NF-κB pathway in lung adenocarcinoma.

doi: 10.1016/j.lfs.2024.122437

Figure Lengend Snippet: Fig.2 Expression of TPX2 in A549 cells affects macrophage polarization. (A) Detection of macrophage surface markers CD80 and CD163 after cell supernatant induction in A549 group, Vehicle group and TPX2-shRNA group. (B) The analysis percentage of macrophage surface marker CD80, CD163; CD80/CD163 after induction by cell supernatant of the three groups. (C) CD80 and CD163 protein expression in macrophages after induction in the supernatant of the three groups cells. (D) The

Article Snippet: THP-1 cell differentiation into macrophages was induced by culturing for 24 h. The supernatant of each group was then applied for 48 h. The induced cells were digested and washed twice, followed by the addition of PE Anti-Human CD80 (B7-1) (PE-65083, Proteintech, China) and APC Anti-Human CD163 (GHI/61) (APC-65169, Proteintech, China) antibodies and resuspended to flow cytometry (BD Biosciences, USA).

Techniques: Expressing, shRNA, Marker