Journal: Journal of Translational Medicine

Article Title: Niraparib exhibits a synergistic anti-tumor effect with PD-L1 blockade by inducing an immune response in ovarian cancer

doi: 10.1186/s12967-021-03073-0

Figure Lengend Snippet: The information of antibodies used for western blot, immunohistochemistry, and flow cytometry

Article Snippet: CD3 and isotype , Human , 5 μl of antibody per test , Elabscience, E-AB-F1001E , Flow cytometry.

Techniques: Western Blot, Immunohistochemistry, Flow Cytometry

Journal: The Journal of Physiology

Article Title: T cells mediate cell non‐autonomous arterial ageing in mice

doi: 10.1113/jp281698

Figure Lengend Snippet: Figure 1. Anti-CD3 F(ab’)2 fragment treatment results in depletion of T cells in the blood and spleen Blood (100 μL) from young isotype (n = 10), young anti-CD3 (n = 10), old isotype (n = 9) and old anti-CD3 (n = 10) mice was directly stained. Spleens from young isotype (n = 11–14), young anti-CD3 (n = 8–11), old isotype (n = 15–17) and old anti-CD3 (n = 18–19) mice were enzymatically digested and passed through a cell strainer and then stained for CD45 (total leukocytes), CD3 (pan T cells), CD4 and CD8. Percentages of CD3+ cells and the CD4-to-CD8 ratio in (A) blood and (B) spleen were assessed by flow cytometry. A two-way ANOVA was employed to assess the effects of age and anti-CD3 treatment, P values for age, treatment and the age × treatment interaction are inset on each panel. Data are shown as means ± standard deviation, n represents the number of independent animals in each group. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Following stimulation, cells were labelled with the following anti-mouse antibodies: PE/Dazzle-CD45, Biolegend #109846, APC-CD3, Tonbo #20-0032, FITC-CD4, Tonbo #30-0041 and PE-Cy5-CD8 Tonbo #55-0081.

Techniques: Staining, Flow Cytometry, Standard Deviation

Journal: The Journal of Physiology

Article Title: T cells mediate cell non‐autonomous arterial ageing in mice

doi: 10.1113/jp281698

Figure Lengend Snippet: Figure 2. Aortic T cell accumulation with age and anti-CD3 F(ab’)2 Fragment treatment Single-cell suspensions of thoracic aortas from young isotype (n = 7–12), young anti-CD3 (n = 9–11), old isotype (n = 8–14) and old anti-CD3 (n = 10–19) mice were stained with antibodies against CD45 (total leukocytes), CD3 (pan T cells), CD4, CD8, CD44 (memory) and CD62L (central vs. effector). A, number of total leukocytes per aorta. B, sample aortic CD4 and CD8 flow cytometry plot. C, aortic pan, CD4 and CD8 T cell counts. D, percentage of CD3 cells out of total aortic immune cells. E, aortic CD4:CD8 ratio. F, sample aortic naïve (CD44lo) vs. memory (CD44hi) flow cytometry plots. G, aortic CD8 effector memory T cell proportion and counts. To assess aortic macrophage and B cell accumulation, thoracic aorta single-cell suspensions were stained for CD45 (total leukocytes), CD19 (B cells), CD64 (macrophages), CD11c (exclusion of dendritic cells) and CD206 (M1/M2 macrophage phenotype. H, aortic B cell counts. I, aortic macrophage counts. J, macrophage M1:M2 ratio. A two-way ANOVA was employed to assess the effects of age and anti-CD3 treatment, P values for age, treatment and the age × treatment interaction are inset on each panel. When a significant age × treatment interaction occurred, Tukey’s post hoc test was employed to determine group differences. Significant post hoc test P values are included on the panel with a horizontal line indicating the group comparison. Data are shown as means ± standard deviation, n represents the number of independent animals in each group. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Following stimulation, cells were labelled with the following anti-mouse antibodies: PE/Dazzle-CD45, Biolegend #109846, APC-CD3, Tonbo #20-0032, FITC-CD4, Tonbo #30-0041 and PE-Cy5-CD8 Tonbo #55-0081.

Techniques: Staining, Flow Cytometry, Comparison, Standard Deviation

Journal: The Journal of Physiology

Article Title: T cells mediate cell non‐autonomous arterial ageing in mice

doi: 10.1113/jp281698

Figure Lengend Snippet: Figure 3. Ageing results in an enhanced proinflammatory phenotype of aortic accumulating T cells Single-cell suspensions of thoracic aortas from young (n = 6) and old (n = 8) mice were activated in vitro and stained for CD45 (total leukocytes), CD3 (pan T cells), CD4, CD8, interferon (IFN)-γ and tumour necrosis factor (TNF)-α. A, sample IFN-γ flow cytometry plots. B, proportion and (C) number of IFN-γ -producing T cells. D, sample TNF-α flow cytometry plots. E, proportion and (F) number of TNF-α-producing T cells. Group differences were assessed with an independent samples t test, P values are included on the panel with a horizontal line indicating the group comparison. Data are shown as means ± standard deviation, n represents the number of independent animals in each group. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Following stimulation, cells were labelled with the following anti-mouse antibodies: PE/Dazzle-CD45, Biolegend #109846, APC-CD3, Tonbo #20-0032, FITC-CD4, Tonbo #30-0041 and PE-Cy5-CD8 Tonbo #55-0081.

Techniques: In Vitro, Staining, Flow Cytometry, Comparison, Standard Deviation

Journal: The Journal of Physiology

Article Title: T cells mediate cell non‐autonomous arterial ageing in mice

doi: 10.1113/jp281698

Figure Lengend Snippet: Figure 4. T cell depletion reverses age-related increases in large artery stiffness Aortic pulse wave velocity (PWV) was assessed in young (n = 19) and old (n = 25) mice (A) before anti-CD3 treatment and an independent samples t test was used to assess group differences. P values are included on the panel with a horizontal line indicating the group comparison. PWV was assessed before and after anti-CD3 F(ab’)2 fragment treatment in (B) young (n = 10 per group) and (C) old mice (n = 12–13 per group). A repeated measures ANOVA was used to assess the effect of time, treatment and time × treatment interaction, with P values inset on each panel. When a significant age × treatment interaction occurred, Sidak’s post hoc test was employed to determine group differences. Significant post hoc test P values are included on the panel with a vertical line indicating the group comparison. D, gene expression of Nox2, Xanthine Oxidase (Xo), superoxide dismutase isoforms 1–3 (Sod1, 2 & 3) from aortas of young isotype (n = 7), young anti-CD3 (n = 7) old isotype (n = 5) and old anti-CD3-treated (n = 5) mice. Gene expression data are expressed as fold change compared with young isotype calculated using the Ct method. Two-way ANOVA was employed to assess the effects of age and anti-CD3 treatment. P values for age, treatment and the age × treatment interaction are inset on each panel. Data are shown as means ± standard deviation, n represents the number of independent animals in each group.

Article Snippet: Following stimulation, cells were labelled with the following anti-mouse antibodies: PE/Dazzle-CD45, Biolegend #109846, APC-CD3, Tonbo #20-0032, FITC-CD4, Tonbo #30-0041 and PE-Cy5-CD8 Tonbo #55-0081.

Techniques: Comparison, Gene Expression, Standard Deviation

Journal: The Journal of Physiology

Article Title: T cells mediate cell non‐autonomous arterial ageing in mice

doi: 10.1113/jp281698

Figure Lengend Snippet: Figure 5. Mesenteric T cell accumulation with age and anti-CD3 F(ab’)2 Fragment treatment Single-cell suspensions of the mesenteric vascular arcade from young isotype (n = 8–14), young anti-CD3 (n = 10–12), old isotype (n = 9–21) and old anti-CD3 (n = 7–11) mice (excluding lymph nodes) were stained for CD45 (total leukocytes), CD3 (pan T cells), CD4, CD8, CD44 (memory) and CD62L (central vs. effector). A, number of total leukocytes per mesentery. B, proportion of CD3 cells out of all mesenteric leukocytes. C, mesenteric CD3 cell counts. D, mesenteric arcade mass. E, mesenteric CD3 cell counts normalized to tissue mass. F, sample CD4 and CD8 flow cytometry plot. G, mesenteric CD4 (left) and CD8 (right) T cell counts. H, mesenteric CD4:CD8 ratio. I, sample mesenteric naïve (CD44lo) vs. memory (CD44hi) flow cytometry plots. J, proportion of mesenteric (left) and counts (right) of CD8 CD44hi/CD62Llo effector memory T cell counts. To assess mesenteric macrophage and B cell accumulation, mesenteric single-cell suspensions were stained for CD45 (total leukocytes), CD19 (B cells), CD64 (macrophages), CD11c (exclusion of dendritic cells) and CD206 (M1/M2 macrophage phenotype). K, mesenteric B cell counts. L, mesenteric macrophage counts. M, macrophage M1:M2 ratio. Two-way ANOVA was employed to assess the effects of age and anti-CD3 treatment. P values for age, treatment and the age × treatment inter- action are inset on each panel. When a significant age × treatment interaction occurred, Tukey’s post hoc test was employed to determine group differences. Significant post hoc test P values are included on the panel with a horizontal line indicating the group comparison. Data are shown as means ± standard deviation, n represents the number of independent animals in each group. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Following stimulation, cells were labelled with the following anti-mouse antibodies: PE/Dazzle-CD45, Biolegend #109846, APC-CD3, Tonbo #20-0032, FITC-CD4, Tonbo #30-0041 and PE-Cy5-CD8 Tonbo #55-0081.

Techniques: Staining, Flow Cytometry, Comparison, Standard Deviation

Journal: The Journal of Physiology

Article Title: T cells mediate cell non‐autonomous arterial ageing in mice

doi: 10.1113/jp281698

Figure Lengend Snippet: Figure 6. Ageing results in an enhanced proinflammatory phenotype of mesenteric accumulating T cells To assess cytokine production, mesenteric single-cell suspensions from young (n = 6) and old (n = 8) mice were activated in vitro. Cells were then stained for CD45 (total leukocytes), CD3 (pan T cells), CD4, CD8, interferon (IFN)-γ and tumour necrosis factor (TNF)-α. A, sample IFN-γ flow cytometry plots. B, proportion and C, number of IFN-γ -producing T cells. D, sample TNF-α flow cytometry plots. E, proportion and (F) number of TNF-α-producing T cells. Group differences were assessed with an independent samples t test. P values are included on the panel with a horizontal line indicating the group comparison. Data are shown as means ± standard deviation, n represents the number of independent animals in each group. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: Following stimulation, cells were labelled with the following anti-mouse antibodies: PE/Dazzle-CD45, Biolegend #109846, APC-CD3, Tonbo #20-0032, FITC-CD4, Tonbo #30-0041 and PE-Cy5-CD8 Tonbo #55-0081.

Techniques: In Vitro, Staining, Flow Cytometry, Comparison, Standard Deviation

Journal: The Journal of Physiology

Article Title: T cells mediate cell non‐autonomous arterial ageing in mice

doi: 10.1113/jp281698

Figure Lengend Snippet: Figure 7. T cell depletion results in augmented endothelium-dependent dilatation in mesenteric arteries from old mice Endothelium-dependent dilatation was assessed in second order mesenteric arteries from (A) young isotype control, young anti-CD3, (B) old isotype control and old anti-CD3 mice in response to increasing doses of acetylcholine. C, endothelium-independent dilatation was assessed in second order mesenteric arteries in from young isotype control, young anti-CD3, old isotype control and old anti-CD3 mice in response to increasing doses of sodium nitroprusside. Dose–response curve data are expressed as means ± standard deviation. A repeated measures ANOVA was used to assess the effect of dose, treatment and dose × treatment interaction, with P values inset on each panel. n sizes are in parentheses next to the corresponding group in each panel legend and represent the number of independent animals in each group.

Article Snippet: Following stimulation, cells were labelled with the following anti-mouse antibodies: PE/Dazzle-CD45, Biolegend #109846, APC-CD3, Tonbo #20-0032, FITC-CD4, Tonbo #30-0041 and PE-Cy5-CD8 Tonbo #55-0081.

Techniques: Control, Standard Deviation

Journal: The Journal of Physiology

Article Title: T cells mediate cell non‐autonomous arterial ageing in mice

doi: 10.1113/jp281698

Figure Lengend Snippet: Figure 8. T cell depletion results in augmented nitric oxide bioavailability in mesenteric arteries from old mice Endothelium-dependent dilatation in response to increasing doses of acetylcholine was assessed in second order mesenteric arteries from old isotype control or old mice treated with anti-CD3 F(ab)’2 fragments in the presence or absence of (A) L-NAME, (B) TEMPOL and (C) TEMPOL & L-NAME. Endothelium-dependent dilatation was also assessed in arteries from young isotype control mice or young mice treated with anti-CD3 F(ab)’2 fragments in the presence or absence of (D) L-NAME, (E) TEMPOL and (F) TEMPOL & L-NAME. Dose–response curve data are expressed as means ± standard deviation. A repeated measures ANOVA was used to assess the effect of dose, group and dose × group interaction, with P values inset on each panel. In the case of a significant group effect, Tukey’s post hoc test was used to compare groups with comparison P values indicated on the legend of each figure panel. In the case of a significant dose × group interaction, Tukey’s post hoc test was employed to assess significant main effects of dose with P values reported in the text. n sizes are in parentheses next to the corresponding group in each panel legend and represent the number of independent animals in each group.

Article Snippet: Following stimulation, cells were labelled with the following anti-mouse antibodies: PE/Dazzle-CD45, Biolegend #109846, APC-CD3, Tonbo #20-0032, FITC-CD4, Tonbo #30-0041 and PE-Cy5-CD8 Tonbo #55-0081.

Techniques: Control, Standard Deviation, Comparison

Journal: Journal of Extracellular Vesicles

Article Title: High‐Yield Outer Membrane Vesicles Derived From Probiotics as a Nanoplatform for Precise Treatment and Prophylaxis of Pseudomonas aeruginosa Infection

doi: 10.1002/jev2.70194

Figure Lengend Snippet: In vivo immune response and preventive effect activated by FI‐mOMVs. (a) Schematic diagram of immunization, sample collection and challenge schedule. BALB/c mice were immunized s.c. with the indicated formulations on Days 0, 14, 28 ( n = 8), and a PBS control was set up ( n = 5). (b) Serum was collected 7, 14, 21, 28, 35 and 42 days after initial vaccination, and the FI antigen–specific IgG antibody titre was detected by ELISA. (c–e) Detection of antigen‐specific cellular immune response. The spleen lymphocytes were isolated 6 weeks after the initial immunization. The proliferation levels were detected by CCK8 (c), and the expression levels of IFN‐γ (d) and IL‐4 (e) in the culture supernatant were determined by ELISA. (f, g) Percentage of CD3 + CD4 + T lymphocytes (f), and CD3 + CD8 + T lymphocytes (g) in the spleen on Day 42 after immunization ( n = 8). Mice without immunization were used as controls ( n = 5). (h) Survival curves of the mice infected with PAO1 (1 × 10 8 CFU/mouse, i.n.) after different treatments ( n = 12). (i, j) Quantification of the number of bacteria in the lung (i) and the pathological changes of the lung (j) of the mice infected with a sublethal dose of PAO1 (5 × 10 6 CFU/mouse, i.n.) after different treatments ( n = 6). Scale bar, 200 µm. The data are presented as mean ± SEM. Statistical analysis was performed by a two‐tailed unpaired Student's t ‐test (b–g), log‐rank (Mantel–Cox) test (h), or two‐tailed Mann–Whitney U ‐test (i), and differences were considered significant at p < 0.05. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001 and **** p < 0.0001.

Article Snippet: The cell suspensions (100 μL) were taken and stained with FITC‐conjugated anti‐mouse CD4 (cat# FITC‐65104, Proteintech, USA), PE‐conjugated anti‐mouse CD8 (cat# PE‐65069, Proteintech, USA) and APC‐conjugated anti‐mouse CD3 (cat# APC‐65077, Proteintech, USA) for 30 min at 37°C in the dark.

Techniques: In Vivo, Control, Enzyme-linked Immunosorbent Assay, Isolation, Expressing, Infection, Bacteria, Two Tailed Test, MANN-WHITNEY