Journal: PLOS Biology

Article Title: Adiponectin in the mammalian host influences ticks’ acquisition of the Lyme disease pathogen Borrelia

doi: 10.1371/journal.pbio.3002331

Figure Lengend Snippet: (A) Histamine concentration in WT ( n = 5) and KO naïve mice ( n = 5), or after tick bite ( n = 10). (B) Injection of Evans blue during tick feeding. The white circles indicated WT ( n = 6) mice have more vascular leakage than KO mice ( n = 6) at the tick bite site. Quantification of Evans blue leakage at the tick bite site of WT ( n = 6) and KO mice ( n = 6). (C) The CD11b + Ly6G + /CD45 + PMN population and the percentage of PMNs in the total CD45 + leukocyte cell population in WT ( n = 4) and KO mice ( n = 4) after tick bite. The CD11b + CD11c − /CD45 + Ly6G − MAC population and the percentage of MACs in the total CD45 + leukocyte cell population in WT ( n = 4) and KO mice ( n = 4) after tick bite. (D) Semiquantitative histopathologic scoring of tick bite/inflammation sites show there is no significant difference in the severity of injury (Injury) but there is an overall increase in the degree of inflammation (Inflammation) in WT mice ( n = 5) compared to adiponectin KO mice ( n = 4). Representative HE-stained sections of WT and adiponectin KO tick bite lesions (arrows). Scale bars = 50 μm, * = subcutis, and ** = ear cartilage. For all the data, statistical significance was assessed using a nonparametric Mann–Whitney test (* p < 0.05; ** p < 0.01; ns, p > 0.05). Data underlying this figure can be found in . HE, hematoxylin and eosin; KO, knock out; MAC, macrophage; PMN, polymorphonuclear neutrophil; WT, wild-type.

Article Snippet: For testing neutrophils, dendritic cells, macrophages, the cells were further incubated with fluorochrome-conjugated monoclonal antibodies against CD45 (PerCP; BD Pharmingen; #561047), CD11b (PE; Biolegend; #101208), CD11c (PE-Cy7; BD Pharmingen; #558079), and Ly6G (FITC; Tonbo; #35–5931) for 30 min at room temperature.

Techniques: Concentration Assay, Injection, Staining, MANN-WHITNEY, Knock-Out

Journal: Science Advances

Article Title: Elevated type I interferon responses potentiate metabolic dysfunction, inflammation, and accelerated aging in mtDNA mutator mice

doi: 10.1126/sciadv.abe7548

Figure Lengend Snippet: ( A and B ) Twelve-month-old WT ( n = 8) and POLG ( n = 7) mice were challenged with LPS [50 mg/kg by intraperitoneal (i.p.) injection]. Kaplan-Meyer survival analysis was performed (A). Plasma cytokine profiles were determined by multianalyte bead-based immunoassay on n ≥ 6 biological samples per group (B). Log-rank (Mantel-Cox) test was used to compare percent survival between different groups. ( C and D ) CD11b + Ly6C hi inflammatory monocyte population in whole blood from 12-month-old WT and POLG mice was evaluated by flow cytometry. Pseudo-color plots are representative of four independent experiments (C), and quantification of the percentage of CD11b + Ly6C hi cells is shown in (D). ( E and F ) CD11b + Ly6G + blood neutrophil population in 12-month-old WT and POLG mice was determined by flow cytometry. Pseudo-color plots are representative of four independent experiments (E), and quantification of the percentage of CD11b + Ly6G + cells is shown in (F). ( G to I ) CD11b + Ly6C hi TNFα + inflammatory monocyte population in unstimulated (C) or LPS-challenged (L) whole blood from 12-month-old WT and POLG mice was evaluated by flow cytometry. Histograms are representative of four independent experiments (G). Quantification of CD11b + Ly6C hi TNFα + mean fluorescent intensity (MFI) is shown in (H), and the percentage of CD11b + Ly6C hi TNFα + cells is shown in (I). ( J to L ) CD11b + Ly6G + TNFα + neutrophil population in unstimulated or LPS-challenged whole blood from 12-month-old WT and POLG mice was evaluated by flow cytometry. Histograms are representative of four independent experiments (J). Quantification of CD11b + Ly6G + TNFα + MFI is shown in (K), and the percentage of CD11b + Ly6G + TNFα + neutrophils is shown in (L). Unless stated, statistical significance was determined using unpaired Student’s t test after Shapiro-Wilk normality test. * P < 0.05, ** P < 0.01, and *** P < 0.001. Error bars represent SEM.

Article Snippet: Antibodies and reagents used for flow cytometry included the following: phycoerythrin/Cy7 anti-mouse TNFα antibody (506324), PerCP/Cy5.5 anti-mouse Ly6C antibody (128012), fluorescein isothiocyanate anti-mouse Ly6C antibody (128006), and PerCP/Cy5.5 anti-mouse IL-12/IL-23 p40 (505211) (from BioLegend); purified anti-mouse CD16 / CD32 (2.4G2) (70-0161), violetFluor 450 anti-mouse Ly6G (1A8) (75-1276), and allophycocyanin anti-human/mouse CD11b (M1/70) (20-0112) (from Tonbo); IL-6 (11-7061-81) (from Invitrogen); IL-1β (31202) (from Cell Signaling Technology); brefeldin A solution (420601) and monensin solution (420701) (from BioLegend); mouse TNFα (430904) and IL-6 (431304) enzyme-linked immunosorbent assay (ELISA) kits (from BioLegend); and Griess Reagent System (G2930) (from Promega).

Techniques: Injection, Clinical Proteomics, Bead-based Assay, Flow Cytometry

Journal: Science Advances

Article Title: Elevated type I interferon responses potentiate metabolic dysfunction, inflammation, and accelerated aging in mtDNA mutator mice

doi: 10.1126/sciadv.abe7548

Figure Lengend Snippet: ( A and B ) Twelve-month-old WT, POLG, POLG cGAS −/− , POLG Sting −/− , and POLG Ifnar −/− ( n = 5 to 8 per group) mice were intraperitoneally injected with LPS (50 mg/kg). Kaplan-Meyer survival analysis was performed (A). Plasma was collected at indicated time points ( n ≥ 6 at 2 hours and n = 4 at 6 hours) and subjected to multianalyte cytokine analysis (B). Statistical comparisons in (B) were made against LPS-injected POLG mice. Log-rank (Mantel-Cox) test was used to compare percent survival between different groups. ( C and D ) CD11b + Ly6C hi inflammatory monocyte population in whole blood from 12-month-old mice was evaluated by flow cytometry. Pseudo-color plots are representative of four independent experiments (C), and quantification of the percentage of CD11b + Ly6C hi cells is shown in (D). ( E and F ) CD11b + Ly6C hi TNFα + inflammatory monocyte population in unstimulated or LPS challenged whole blood from 12-month-old cohorts was evaluated by flow cytometry. Quantification of CD11b + Ly6C hi TNFα + MFI is shown in (E), and the percentage of CD11b + Ly6C hi TNFα + cells is shown in (F). Unless stated, statistical significance was determined using analysis of variance (ANOVA) and Tukey post hoc test. * P < 0.05, ** P < 0.01, and *** P < 0.001. Error bars represent SEM.

Article Snippet: Antibodies and reagents used for flow cytometry included the following: phycoerythrin/Cy7 anti-mouse TNFα antibody (506324), PerCP/Cy5.5 anti-mouse Ly6C antibody (128012), fluorescein isothiocyanate anti-mouse Ly6C antibody (128006), and PerCP/Cy5.5 anti-mouse IL-12/IL-23 p40 (505211) (from BioLegend); purified anti-mouse CD16 / CD32 (2.4G2) (70-0161), violetFluor 450 anti-mouse Ly6G (1A8) (75-1276), and allophycocyanin anti-human/mouse CD11b (M1/70) (20-0112) (from Tonbo); IL-6 (11-7061-81) (from Invitrogen); IL-1β (31202) (from Cell Signaling Technology); brefeldin A solution (420601) and monensin solution (420701) (from BioLegend); mouse TNFα (430904) and IL-6 (431304) enzyme-linked immunosorbent assay (ELISA) kits (from BioLegend); and Griess Reagent System (G2930) (from Promega).

Techniques: Injection, Clinical Proteomics, Flow Cytometry