Journal: Cells

Article Title: Statins Modulate Microenvironmental Cues Driving Macrophage Polarization in Simulated Periodontal Inflammation

doi: 10.3390/cells12151961

Figure Lengend Snippet: MyD88−dependent modulation of M2 polarization by simvastatin. ( A ) A schematic representation of simvastatin, MyD88 inhibitor treatment, and THP-1-macrophages (M0–M1–M2). ( B ) Cell surface antigen analysis of M2 macrophage markers CD68, CD163, and CD206 was conducted using flow cytometry. Expression levels were normalized using IgG isotype control, and values were expressed in percentage of positive cells (mean ± SD) from three independent preparations. ( C ) RT-PCR analysis and heat maps summarizing TLR and NFkB signaling from M0 (CCL2), M1 (LPS, IFNγ, TNFα), and M2 (IL4, IL10, TGFβ) macrophages. (n = 3) preparations per treatment group, and data found to be significantly different were plotted as heat maps. ( D ) Graphical summary of statin-mediated suppression of TLR and NFkB signaling.

Article Snippet: Phenotyping of macrophages was performed in 1% BSA and 3% human serum PBS according to standard methods using a panel of antibodies targeting CD68 (R and D Systems Cat# IC20401P, Minneapolis, MN, USA, RRID: http://scicrunch.org/resolver/AB_2074835 , accessed on 28 April 2023), CD163 (R and D Systems Cat# FAB1607P, RRID: http://scicrunch.org/resolver/AB_2074536 , accessed on 28 April 2023), and CD206 (R and D Systems Cat# FAB25342P, RRID: http://scicrunch.org/resolver/AB_10889015 , accessed on 28 April 2023); antibodies all from R and D Systems.

Techniques: Flow Cytometry, Expressing, Control, Reverse Transcription Polymerase Chain Reaction

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: Vaccines

Article Title: Simultaneous Administration of Recombinant Measles Viruses Expressing Respiratory Syncytial Virus Fusion (F) and Nucleo (N) Proteins Induced Humoral and Cellular Immune Responses in Cotton Rats

doi: 10.3390/vaccines7010027

Figure Lengend Snippet: The proportion of CD8 + /IFN-γ + cells in spleen cells obtained four days after the RSV challenge (Day 88), stimulated with UV-inactivated RSV Long strain, F, NP, and M2-1 peptides. Spleen cells were obtained after the RSV challenge and analyzed for IFN-γ production by CD8 + cells. The bar represents the mean of 3~4 rats per group ± SEM. (* p < 0.05).

Article Snippet: After the stimulation, splenocytes were washed and incubated with an anti-CD8 antibody (R&D Systems, Minneapolis, MN, USA) at 4 °C for 30 min. Splenocytes were fixed with fixation buffer in the Cytofix/cytoperm kit, and intracellular cytokines were stained with a goat IgG antibody against IFN-γ (R&D Systems, USA) and anti-goat IgG PE-Cy7 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4 °C for 60 min. Cellular populations were analyzed using flow cytometry with Cytomics FC 500 (Beckman Coulter, Inc., Indianapolis, IN, USA) and counted until 100,000 cells.

Techniques:

Journal: Vaccines

Article Title: Simultaneous Administration of Recombinant Measles Viruses Expressing Respiratory Syncytial Virus Fusion (F) and Nucleo (N) Proteins Induced Humoral and Cellular Immune Responses in Cotton Rats

doi: 10.3390/vaccines7010027

Figure Lengend Snippet: Detection of IFN-γ + expressing cells in lung homogenates. Lung homogenates were obtained after the RSV challenge on Day 88. IFN-γ-producing cells were separated further into CD8 + and CD4 + cells. The bar represents the mean of 3~4 rats per group ± SEM. (* p < 0.05).

Article Snippet: After the stimulation, splenocytes were washed and incubated with an anti-CD8 antibody (R&D Systems, Minneapolis, MN, USA) at 4 °C for 30 min. Splenocytes were fixed with fixation buffer in the Cytofix/cytoperm kit, and intracellular cytokines were stained with a goat IgG antibody against IFN-γ (R&D Systems, USA) and anti-goat IgG PE-Cy7 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4 °C for 60 min. Cellular populations were analyzed using flow cytometry with Cytomics FC 500 (Beckman Coulter, Inc., Indianapolis, IN, USA) and counted until 100,000 cells.

Techniques: Expressing