Journal: Brain Sciences

Article Title: Potential for Cell-Mediated Immune Responses in Mouse Models of Pelizaeus-Merzbacher Disease

doi: 10.3390/brainsci3041417

Figure Lengend Snippet: White matter microglia/macrophages are activated in the dorsal funiculi of male Plp1 mutant mice. Cervical spinal cord sections from ( A ) mildly-affected P16 rsh ( B ) severely-affected P16 msd ( C ) subclinical P25 Plp1 #66 heterozygous ( D ) moderately-affected P25 Plp1 #66 homozygous mice labeled with antibodies against Iba-1 (red) and CD68 (green). Many Iba-1 positive microglia/macrophages with swollen cell bodies and short processes are labeled with anti-CD68 antibodies in rsh and msd mice, which is indicative of activated cells. The insets in these panels show the canonical activated morphology revealed with anti-Iba-1 antibodies and strong CD68 staining. In contrast, there is little evidence for broad CD68 labeling in Plp1 #66 heterozygous mice in panel C, which is consistent with previous data that immune cells are rarely activated . Activation of microglia/macrophages is apparent for Plp1 #66 homozygous mice in panel D, which exhibit an intermediate level of CD68 labeling. Scale bar: 95 μm, insets 15 μm.

Article Snippet: Antibodies used were: mouse anti-MBP (1:1000, SMI99, Sternberger Monoclonal Inc., Baltimore, MD, USA); rabbit anti-Iba-1; monoclonal anti-human CD68 antibody (clone 514H12, AbD Serotec, Oxford, UK).

Techniques: Mutagenesis, Labeling, Staining, Activation Assay

Journal: Brain Sciences

Article Title: Potential for Cell-Mediated Immune Responses in Mouse Models of Pelizaeus-Merzbacher Disease

doi: 10.3390/brainsci3041417

Figure Lengend Snippet: Proportion of cervical spinal cord Iba-1 + microglia/macrophages in dorsal column, ventral column and lateral funiculus that are activated (CD68 + ).

Article Snippet: Antibodies used were: mouse anti-MBP (1:1000, SMI99, Sternberger Monoclonal Inc., Baltimore, MD, USA); rabbit anti-Iba-1; monoclonal anti-human CD68 antibody (clone 514H12, AbD Serotec, Oxford, UK).

Techniques:

Journal: Brain Sciences

Article Title: Potential for Cell-Mediated Immune Responses in Mouse Models of Pelizaeus-Merzbacher Disease

doi: 10.3390/brainsci3041417

Figure Lengend Snippet: Oligodendrocytes in gray matter from P16 rsh and msd mice do not undergo a UPR and microglia/macrophages exhibit resting morphology. ( A – C ) Iba1 (red) and CD68 (green) antibody labeling in dorsal spinal cord white matter tracts of P16 wild type ( A ) rsh ( B ) and msd ( C ) mice. Although the morphology and CD68 staining is characteristic of resting microglia/macrophages in wild type mice (white arrowheads), these cells are activated in the Plp1 mutants. ( D – F ) In contrast to white matter regions, microglia/macrophages have a resting state phenotype in the adjacent substantia gelatinosa (gray matter) from wild type and mutant mice (black arrowheads). ( G – I ) A major difference between these regions in rsh and msd mice is the relative abundance of oligodendrocytes undergoing an unfolded protein response (UPR), which is evident by the large number of CHOP + cells (green) in white matter (above the dotted line) compared to gray matter (below). We have previously shown that 100% of CHOP + cells in these mutants are oligodendrocytes . Dotted lines mark the white/gray matter boundary. Scale bar in I: 100 μm.

Article Snippet: Antibodies used were: mouse anti-MBP (1:1000, SMI99, Sternberger Monoclonal Inc., Baltimore, MD, USA); rabbit anti-Iba-1; monoclonal anti-human CD68 antibody (clone 514H12, AbD Serotec, Oxford, UK).

Techniques: Antibody Labeling, Staining, Mutagenesis

Journal: Brain Sciences

Article Title: Potential for Cell-Mediated Immune Responses in Mouse Models of Pelizaeus-Merzbacher Disease

doi: 10.3390/brainsci3041417

Figure Lengend Snippet: Activation of rsh and msd microglia/macrophages in optic nerve at P16. ( A ) Longitudinal section from wild type mouse reveals Iba-1 + microglia/macrophages (red) with non-activated morphology (arrowheads). Most of these cells do not express CD68 or express the protein at low levels (green). ( B ) DAPI staining showing the nuclei of the microglia/macrophages in ( A ). ( C , E ) Iba-1 + microglia/macrophages from rsh ( C ) and msd ( E ) mice exhibit an activated morphology with enlarged cell bodies and thickened processes. Most of these cells express CD68 at high levels, which is localized to perinuclear regions. ( D , F ) DAPI staining of the fields in ( C , E ). Scale bar in F: 30 μm.

Article Snippet: Antibodies used were: mouse anti-MBP (1:1000, SMI99, Sternberger Monoclonal Inc., Baltimore, MD, USA); rabbit anti-Iba-1; monoclonal anti-human CD68 antibody (clone 514H12, AbD Serotec, Oxford, UK).

Techniques: Activation Assay, Staining

Journal: Brain Sciences

Article Title: Potential for Cell-Mediated Immune Responses in Mouse Models of Pelizaeus-Merzbacher Disease

doi: 10.3390/brainsci3041417

Figure Lengend Snippet: Expression fold changes of other non-chromosome 17 interferon-induced genes in microarray data for rsh and msd mice.

Article Snippet: Antibodies used were: mouse anti-MBP (1:1000, SMI99, Sternberger Monoclonal Inc., Baltimore, MD, USA); rabbit anti-Iba-1; monoclonal anti-human CD68 antibody (clone 514H12, AbD Serotec, Oxford, UK).

Techniques: Expressing, Microarray

Journal: Clinics

Article Title: M2-Polarized tumor-associated macrophages are associated with poor prognoses resulting from accelerated lymphangiogenesis in lung adenocarcinoma

doi: 10.1590/S1807-59322011001100006

Figure Lengend Snippet: M2- and M1-polarized TAMs in lung adenocarcinoma. (A) The coexpression of CD68 and MMR in TAMs in human lung adenocarcinoma sections. (B) The coexpression of CD68 and iNOS in TAMs in human lung adenocarcinoma sections.

Article Snippet: Next, slides were immunostained using a monoclonal mouse anti-human CD68 antibody (1∶100; Zhongshan Company, Beijing, China).

Techniques:

Journal: BMC Cancer

Article Title: Local iron homeostasis in the breast ductal carcinoma microenvironment

doi: 10.1186/s12885-016-2228-y

Figure Lengend Snippet: FPN1-expressing leukocytes in carcinomas are predominantly M2-like. Sections of normal breast tissue, DCIS (ductal carcinoma in situ) and IDC (invasive ductal carcinoma) to reveal the presence of cells of the macrophage lineage (CD68), and its classical polarization phenotypes, M1-like (CD80) and M2-like (CD163), in FPN1-expressing leukocyte infiltrate. For details see Materials and Methods (Original magnification × 100- upper CD68 images, ×400- squared image series)

Article Snippet: Immunohistochemical staining was performed in 2 μm-thick TMA sections with the following antibodies: rabbit polyclonal anti-human hepcidin-25 antibody (dilution 1:500, Abcam, Cambridge, UK [ ]), rabbit polyclonal anti-human ferroportin 1 antibody (FPN—1:500, Novus Biologicals Europe, Cambridge, UK [ ]), rabbit polyclonal anti-human ferritin antibody (FT—1:1000, Sigma-Aldrich, MO, USA [ ]), mouse monoclonal anti-human CD71 (TFR1 [clone 10 F11]- 1:80, Novocastra, Newcastle, UK [ ]), mouse monoclonal anti-human CD68 (clone Kp-1, 1:2000, A. Menarini Diagnostics, CA, USA), mouse monoclonal anti-human CD163 (clone MRQ-26, 1:100, Cell Marque, CA, USA), mouse monoclonal anti-human CD80 (37711, 1:100, R&D Systems, MN, USA), rabbit polyclonal anti-human CD4 (clone H-370, 1:250, Santa Cruz Biotechnology, TX, USA) and mouse monoclonal anti-human CD8 (clone C8/144B, 1:100, Cell Marque, CA, USA).

Techniques: Expressing, In Situ

Journal: BMC Cancer

Article Title: Local iron homeostasis in the breast ductal carcinoma microenvironment

doi: 10.1186/s12885-016-2228-y

Figure Lengend Snippet: Representative images of the FPN1 analysis by Imaging Flow Cytometry in breast cancer core biopsies. Epithelial cells (EC) are stained by an anti-cytokeratin (CK) FITC. T lymphocytes (T Ly) were identifiable by CD3 PerCP-Cy5.5, B lymphocytes (B Ly) by CD20 PE-Cy7 and macrophages (M0) by CD68 PE-Cy7 (Original magnification × 400)

Article Snippet: Immunohistochemical staining was performed in 2 μm-thick TMA sections with the following antibodies: rabbit polyclonal anti-human hepcidin-25 antibody (dilution 1:500, Abcam, Cambridge, UK [ ]), rabbit polyclonal anti-human ferroportin 1 antibody (FPN—1:500, Novus Biologicals Europe, Cambridge, UK [ ]), rabbit polyclonal anti-human ferritin antibody (FT—1:1000, Sigma-Aldrich, MO, USA [ ]), mouse monoclonal anti-human CD71 (TFR1 [clone 10 F11]- 1:80, Novocastra, Newcastle, UK [ ]), mouse monoclonal anti-human CD68 (clone Kp-1, 1:2000, A. Menarini Diagnostics, CA, USA), mouse monoclonal anti-human CD163 (clone MRQ-26, 1:100, Cell Marque, CA, USA), mouse monoclonal anti-human CD80 (37711, 1:100, R&D Systems, MN, USA), rabbit polyclonal anti-human CD4 (clone H-370, 1:250, Santa Cruz Biotechnology, TX, USA) and mouse monoclonal anti-human CD8 (clone C8/144B, 1:100, Cell Marque, CA, USA).

Techniques: Imaging, Flow Cytometry, Staining