Journal: Frontiers in Immunology

Article Title: Halted Lymphocyte Egress via Efferent Lymph Contributes to Lymph Node Hypertrophy During Hypercholesterolemia

doi: 10.3389/fimmu.2019.00575

Figure Lengend Snippet: Dyslipidemia induces LN lymphangiogenesis (B) LN sections from WT and apoE −/− mice at 22 to 28 weeks of age were stained for B220 and LYVE-1. Images are representative of four independent experiments ( n = 3–4 mice per group). (A) LN cells were stained for CD45, podoplanin and CD31 to identify LECs by FACS analysis. The number of LEC was enumerated in LNs from WT and apoE −/− mice at 22 to 28 weeks of age and expressed as fold change over WT mice. Data is pooled from three independent experiments with 3–5 mice per group in each experiment; * p < 0.05. (C) LYVE-1 + vessels were analyzed for co-expression of the proliferative marker KI67. Arrows indicate co-localization of lymphatics with Ki67. Ki67 bright dividing lymphocytes were also observed. Images are representative of three to four independent experiments ( n = 3–4 mice per group).

Article Snippet: Antibodies used included the following: rat anti-mouse CD31 (Serotec) detected with anti-rat-APC, hamster anti-mouse podoplanin (clone 8.1.1; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA) detected with anti-hamster PE and PerCPcy5.5—conjugated anti-mouse CD45.2 (BD Biosciences).

Techniques: Staining, Expressing, Marker

Journal: Frontiers in Immunology

Article Title: Halted Lymphocyte Egress via Efferent Lymph Contributes to Lymph Node Hypertrophy During Hypercholesterolemia

doi: 10.3389/fimmu.2019.00575

Figure Lengend Snippet: The extended lymphatic network in apoE −/− mice LNs exhibits structural abnormalities. (A) Immunoreactivity for LYVE-1 was examined in LN sections from WT and apoE −/− mice at 22–28 weeks of age. (B) Lumen area of cortical and medullary sinuses was determined on LN sections from 22–28 week-old WT and apoE −/− mice. n = 4 mice per group; *** p < 0.0005 (C) Immunoreactivity for LYVE-1 and B220, (D) LYVE-1 and collagen (Col) type IV or (E) LYVE-1 and CD31 was examined in LN sections from apoE −/− mice at 22–28 weeks of age. Images are representative of 3–4 independent experiments ( n = 3–5 mice per group). Arrows indicate loss of LYVE-1 expression.

Article Snippet: Antibodies used included the following: rat anti-mouse CD31 (Serotec) detected with anti-rat-APC, hamster anti-mouse podoplanin (clone 8.1.1; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA) detected with anti-hamster PE and PerCPcy5.5—conjugated anti-mouse CD45.2 (BD Biosciences).

Techniques: Expressing

Journal: bioRxiv

Article Title: Demonstration of chemotherapeutic mediated lymphatic changes in meningeal lymphatics in vitro, ex vivo, and in vivo

doi: 10.1101/2024.01.06.574460

Figure Lengend Snippet: LN LECs are seeded on the underside of a tissue culture insert for 2 h before inversion and culture for 24 h. Then, meningeal cells are added into the tissue culture insert and cultured an additional 36 h. (A). Suitability of co-culture media is verified via total meningeal cells per field of view (B) and viability of meningeal cells in Vasculife (C). Permeability is quantified via dextran transport assay (D). Representative images of each monolayer are shown, with meningeal cells stained for F-actin (magenta) and LECs stained for CD31 (red). Nuclei, stained with DAPI, are shown in cyan (E). Data shown are biological replicates (n=3), mean +/− SEM, with * denoting p<0.05. Scale bar is 50 µm.

Article Snippet: Membranes were stained with sheep anti-human CD31 (R&D) at a 1:40 dilution and mouse anti-human alpha smooth muscle actin (Abcam) at a 1:200 dilution for 2 h at room temperature (RT) followed by donkey anti-sheep Alexa Fluor 647 secondary antibody (Invitrogen) at a 1:400 dilution and donkey anti-mouse Alexa Fluor 555 antibody (Invitrogen) at a 1:1000 dilution for 1 h at RT.

Techniques: Cell Culture, Co-Culture Assay, Permeability, Transport Assay, Staining

Journal: bioRxiv

Article Title: Demonstration of chemotherapeutic mediated lymphatic changes in meningeal lymphatics in vitro, ex vivo, and in vivo

doi: 10.1101/2024.01.06.574460

Figure Lengend Snippet: ML models are treated with 1 µM carboplatin, 1 µM docetaxel, or vehicle (DMSO) for 24 hours. Representative images of ML models are shown, with nuclei stained with DAPI (cyan), hMCs stained for F-actin (magenta) and LECs stained for CD31 (red). Scale bar is 50 µm (A). Coverage of hMcs (B) and LECs (C) is quantified in ImageJ and reported as percentage, within the ML model and as monocultures. Data shown are biological replicates (n=3) and mean, with * denoting p<0.05, ** denoting p<0.01. a denotes significant difference from docetaxel treatment.

Article Snippet: Membranes were stained with sheep anti-human CD31 (R&D) at a 1:40 dilution and mouse anti-human alpha smooth muscle actin (Abcam) at a 1:200 dilution for 2 h at room temperature (RT) followed by donkey anti-sheep Alexa Fluor 647 secondary antibody (Invitrogen) at a 1:400 dilution and donkey anti-mouse Alexa Fluor 555 antibody (Invitrogen) at a 1:1000 dilution for 1 h at RT.

Techniques: Staining

Journal: Frontiers in Immunology

Article Title: Novel Role for PD-1:PD-L1 as Mediator of Pulmonary Vascular Endothelial Cell Functions in Pathogenesis of Indirect ARDS in Mice

doi: 10.3389/fimmu.2018.03030

Figure Lengend Snippet: Representative dot-plots illustrating how hemorrhagic shock increased the percentage of PD-1 + Ly6G + neutrophils in peripheral blood (A,B) and PD-L1 + CD31 + endothelial cells in lung tissue (C,D) . Summary data document that percentage of PD-1 + Ly6G + neutrophils increases early and significantly at 6 h and continues to increase over the 24 h following Hem (E) . In contrast, the percentage PD-L1 + CD31 + endothelial cells increase at 6 h post-Hem but decreases by 24 h post-Hem (F) . Mean ± SEM, N = 3/group, * p < 0.05 vs. Sham Hem; # p < 0.05 vs. 6 h. One-way ANOVA and a Tukey's multiple comparisons test.

Article Snippet: Cells were incubated with PE-Cy7 labeled rat anti-mouse CD31 and BV421 labeled hamster anti-mouse ICAM-1 (Miltenyi biotec).

Techniques:

Journal: Frontiers in Immunology

Article Title: Novel Role for PD-1:PD-L1 as Mediator of Pulmonary Vascular Endothelial Cell Functions in Pathogenesis of Indirect ARDS in Mice

doi: 10.3389/fimmu.2018.03030

Figure Lengend Snippet: ICAM-1 expression on CD31 + ECs isolated from lung tissue of PD-1 −/− , PD-L1 −/− , and WT Control mice following SH/C and H/C. Mean ± SEM, N = 8/group; @ p < 0.05 vs. Control H/C; * p < 0.05 vs. respective SH/C. One-way ANOVA and a Tukey's Multiple comparisons test.

Article Snippet: Cells were incubated with PE-Cy7 labeled rat anti-mouse CD31 and BV421 labeled hamster anti-mouse ICAM-1 (Miltenyi biotec).

Techniques: Expressing, Isolation, Control