Journal:

Article Title: Trafficking of a Dual-Modality Magnetic Resonance and Fluorescence Imaging Superparamagnetic Iron Oxide-Based Nanoprobe to Lymph Nodes

doi: 10.1007/s11307-010-0424-8

Figure Lengend Snippet: Confocal microscopy of SCION(AlexaFluor555) localization in politeal lymph node 24 hr post-intradermal injection in hind footpad. Panels b-e, g, and i are magnified regions of interest from LNs panels a, f, and h which capture the LNs as a whole or from their circumference. In all nodes, SCION was primarily found in the subcapsular space, trabelculae, and near the exiting efferent vessel in the medullary and hilum regions. (a-e) B6-albino mouse co-injected with FITC-dextran that localized in CD169+ subcapsular macrophages, whereas SCION particle was observed in LYVE-1+ endothelial cells lining the subcapsular floor (b-d) and hilum (a,e). (f-g) CD11c-EYFP mouse showed minimal uptake of SCION by CD11c+ dendritic cells. (h-i) lys-EGFP mouse biopsy found minimal overlap between SCION and lys+ myelomonocytic cells (neutrophil granulocytes and macrophages)

Article Snippet: Sections were blocked in PBS containing 10% goat serum (Jackson Immuno-Research Laboratories) and stained with unconjugated rat anti-mouse CD169 (Cedarlane, CL89149B) and rabbit anti-LVYE1 (Novus Biologicals).

Techniques: Confocal Microscopy, Injection

Journal: Oncoimmunology

Article Title: Stromal niche communalities underscore the contribution of the matricellular protein SPARC to B-cell development and lymphoid malignancies

doi: 10.4161/onci.28989

Figure Lengend Snippet: Stromal-SPARC influence on splenic B-cell maturation. Spleen cells were stained with fluorophore-conjugated antibodies against the indicated marker and analyzed by flow cytometry. ( A ) Confocal microscopy analysis for SPARC, CD29, collagen type-4 (Col4) and CD93 expression in spleens from wild-type (WT) and Sparc −/− mice. ( B–C ) Flow cytometric analysis showing the fraction of B220 + CD93 + ( B ) and B220 + CD93 - ( C ) cells in the spleen of Sparc −/− in comparison to WT mice. ( D–E ) Immature B220 + CD93 + B cells were classified by immunostaining and cytofluorimetric analysis into transitional T1, T2, and T3 cells based on their expression of CD23 and IgM (T1 = IgM + CD23 - , T2 = IgM + CD23 + , T3 = IgM low CD23 + ). ( D ) Collective data for transitional T1, T2 and T3 cells obtained from the analysis of spleen cell suspensions from 5 WT and 5 Sparc −/− mice (n = 5 mice/group; Statistical analysis performed by Student’s t test; * P < 0.05; ** P < 0.01; one representative experiment out of 3). ( E ) Representative contour plots for transitional T1, T2 and T3 cells in WT and Sparc −/− mice. ( F ) Marginal zone B (MZB) cells were defined within the gate of B220 + CD23 - IgD - based on their of high level of CD21/35 and IgM. ( G ) Cumulative data showing the frequency of MZB cells in WT and Sparc −/− (n = 5 mice/group; one representative experiment out of 3) ( H ) Quantitative RT-PCR analysis for deltex-1 ( Dtx-1) mRNA expression performed on total CD19 + cells sorted from the spleen of WT and Sparc −/− mice (n = 3 mice/group). ( I ) Delta-like protein 1 (Dll-1) expression analyzed by flow cytometry in the different myeloid cell subsets defined by the markers CD11b, F4/80 Gr-1 and CD169. (One representative experiment out of 3.) ( J ) Confocal microscopy analysis showing neutrophils localizing to the marginal area, defined by CD169 + marginal zone macrophages, and contacting IgM high MZB cells.

Article Snippet: The following primary monoclonal Abs (mAb) were used for triple IF staining on mouse tissues: APC- rat anti-mouse GR-1 (e-Biosciences); Alexa-546-conjugated rat anti-mouse IgM (Invitrogen); FITC-conjugated CD93: rat anti-mouse CD169 (e-Biosciences); goat anti-mouse SPARC/osteonectin (AF952, R&D System).

Techniques: Staining, Marker, Flow Cytometry, Confocal Microscopy, Expressing, Immunostaining, Quantitative RT-PCR

Journal: Theranostics

Article Title: Abraxane-induced bone marrow CD11b + myeloid cell depletion in tumor-bearing mice is visualized by μPET-CT with 64 Cu-labeled anti-CD11b and prevented by anti-CSF-1

doi: 10.7150/thno.49421

Figure Lengend Snippet: Inhibition of Abraxane-mediated recruitment of macrophages to the tumor by αCSF-1. ( A ) 64 Cu-αCD11b µPET-CT of MDA-MB-435 tumor-bearing female nude mice treated with the single-dose Abraxane regimen as shown in Figure A. Images were acquired 24 h after radiotracer injection. Red arrows: bone marrow of thoracic spine; green arrows: bone marrow of sternum; gold circles: tumor. ( B ) Quantitative analysis of tumor uptake of 64 Cu-αCD11b from images acquired 24 h after radiotracer injection. ( C ) Representative dot plots of CD45 + cells in the tumors stained with CD11b and CD169. ( D ) Quantification of CD11b + CD169 + macrophages in CD45 + cells in the tumors. Data are expressed as mean ± standard deviation (n = 3/group). *, p < 0.05; **, p < 0.01.

Article Snippet: Rat anti-mouse CD11b (clone M1/70; αCD11b) and its phycoerythrin conjugate, rat anti-mouse CD45 FITC (clone 104), rat anti-mouse CD169 eFluor660 (clone SER-4), rat anti-mouse Ly6C allophycocyanin (clone hk1.4), rat anti-mouse Ly6G (Gr-1) PerCP.Cyanine5.5 (clone RB6-8C5), and rat anti-mouse CD127 APC-eFluor780 (clone A7R34) were purchased from eBioscience, Inc (San Diego, CA).

Techniques: Inhibition, Injection, Staining, Standard Deviation

Journal: PLoS ONE

Article Title: Contrasting Roles of Islet Resident Immunoregulatory Macrophages and Dendritic Cells in Experimental Autoimmune Type 1 Diabetes

doi: 10.1371/journal.pone.0150792

Figure Lengend Snippet: Frozen pancreas sections from C57BL/6 mice (n = 5) were stained with a triple antibody cocktail reactive to glucagon, somatostatin, and pancreatic polypeptide (Light Blue); anti-TIM-4 (Green); anti-CD169 (Red); and Hoechst (Blue). White arrows indicate CD169+TIM-4+ cells. Individual panels are shown in . Photomicrographs were taken using 40x objective magnification. The bottom panel shows an enlarged area of the photomicrograph in the top panel.

Article Snippet: Pancreases were snap-frozen; stained with rat anti-mouse CD169, goat anti-mouse TIM-4 (R&D Systems, Minneapolis, MN) and a triple antibody cocktail containing rabbit anti-human glucagon, somatostatin, and pancreatic polypeptide.

Techniques: Staining

Journal: PLoS ONE

Article Title: Contrasting Roles of Islet Resident Immunoregulatory Macrophages and Dendritic Cells in Experimental Autoimmune Type 1 Diabetes

doi: 10.1371/journal.pone.0150792

Figure Lengend Snippet: Purified C57BL/6 pancreatic islets were enzymatically digested and stained with anti-CD45, anti-CD11c, anti-Ly6C, anti-F4/80, and anti-CD16/32. (A) Identification of live CD45+ (not shown) CD11c+Ly6C- (left panel) cells that exhibit an islet-resident macrophage (IRM, F4/80+CD16/32+) or DC (IRDC, F4/80-CD16/32-) phenotype (right panel) are shown. (B) FACS-sorted IRMs and IRDCs were analyzed for Csf1r transcript levels by real-time PCR. Data from independent experiments (n = 4 per group) and their mean are plotted; groups were compared with an unpaired t test. (C) Histograms show protein expression of CD169, TIM-4, and CX3CR1 on gated IRM (solid line) and IRDC (dashed line) subsets. Shaded histograms are the fluorescence minus one (FMO) controls. Geometric mean fluorescence intensities (MFIs) are indicated parenthetically. Data are representative of three independent experiments.

Article Snippet: Pancreases were snap-frozen; stained with rat anti-mouse CD169, goat anti-mouse TIM-4 (R&D Systems, Minneapolis, MN) and a triple antibody cocktail containing rabbit anti-human glucagon, somatostatin, and pancreatic polypeptide.

Techniques: Purification, Staining, Real-time Polymerase Chain Reaction, Expressing, Fluorescence