anti-mouse macrophages/ cd169 (clone moma-1) biotin  (Cedarlane)

Journal: BMC Immunology

Article Title: Autoreactive marginal zone B cells enter the follicles and interact with CD4 + T cells in lupus-prone mice

doi: 10.1186/1471-2172-12-7

Figure Lengend Snippet: Intrafollicular location of MZB cells in B6.TC spleens . A . Representative spleen sections from 3 mo old B6.TC (left) and B6 (right) mice stained with Moma-1-FITC, CD1d-PE and B220-PB. The B220 + CD1d + MZB cells show as bright pink while the other B cells show as blue. The ring of Moma-1 + metallophillic macrophages delineates the MZ inner edge. B . Percentage of CD1d + B220 + B cells relative to total B220 + B cells outside (MZ) and inside (FO) the Moma-1 + ring in 3 mo and 10 mo old B6 and B6.TC mice. The data show means and standard errors of the mean (SEM) calculated of 4 MZ and FO areas for each mouse. ***: p < 0.001 for t tests. C . Representative spleen sections from 10 mo old B6.TC (left) and B6 (right) mice stained with CD4-FITC, CD1d-PE and B220-PB. Boxed areas show multiple contacts between green B6.TC CD4 T cells and pink MZB cells, but not in the B6 spleen. Original magnification: 200X.

Article Snippet: Immunofluorescence staining was performed on frozen sections as previously described [ ], with Moma-1-FITC (Serotec) or CD4-FITC (GK1.5), CD1d-PE (1B1), and IgM a -Biotin-SA-PB or B220-PB (RA3-6B2).

Techniques: Staining

Journal: BMC Immunology

Article Title: Autoreactive marginal zone B cells enter the follicles and interact with CD4 + T cells in lupus-prone mice

doi: 10.1186/1471-2172-12-7

Figure Lengend Snippet: Intrafollicular location of 56R and AM14 HC Tg MZB cells . Representative spleen sections of B6. Sle2 .56R and B6.56R ( A , 100X) and B6.TC.AM14.IgH a/b and B6.AM14.IgH a/b (B , 200X ) mice stained with Moma-1-FITC, CD1d-PE, and IgM a -biotin-SA-PB . C. Quantitation in pixels corresponding to the stain combination specific for each cell type of CD1d + IgM a cells in the MZ (pink) and corresponding FO (blue) B cell areas in B6. Sle2 .56R, B6.56R, B6.TC.AM14.IgH a/b and B6.AM14.IgH a/b mice. Paired MZ and FO values within a strain were compared with the Wilcoxon signed ranked test, and the B6.TC.AM14.IgH a/b and B6.AM14.IgH a/b MZ values were compared with the Mann-Whitney test. CD21 hi CD23 hi representing the T2 and MZB precursor cells expressed as the percentage of transgenic IgM a cells (D) , and percentage of CD21 hi CD23 lo MZ B cells expressed as the percentage of transgenic IgM a cells ( E) and their absolute numbers ( F) in B6. Sle2 .56R and B6.56R and B6.TC.AM14.IgH a/b and B6.AM14.IgH a/b mice. Graphs show means and SEMs, and the statistical significance of Student t tests. *: p < 0.05, **: p < 0.01, ***: p < 0.001.

Article Snippet: Immunofluorescence staining was performed on frozen sections as previously described [ ], with Moma-1-FITC (Serotec) or CD4-FITC (GK1.5), CD1d-PE (1B1), and IgM a -Biotin-SA-PB or B220-PB (RA3-6B2).

Techniques: Staining, Quantitation Assay, MANN-WHITNEY, Transgenic Assay

Journal: PLoS ONE

Article Title: Notch-RBP-J-Independent Marginal Zone B Cell Development in IgH Transgenic Mice with V H Derived from a Natural Polyreactive Antibody

doi: 10.1371/journal.pone.0038894

Figure Lengend Snippet: (A) and (B) Two months following BM transplantation, the four groups of mice were sacrificed and spleen cells were stained using anti-CD19, anti-CD21 and anti-CD23 (A), or anti-CD19, anti-CD21 and anti-CD24 (B) and then analyzed using FCM. The cells were initially gated using anti-CD19. CD24 hi CD21 hi cells represent MZ/T2 B cells, and CD21 med CD24 hi cells represent T1 cells. The numbers denote the percentages of cells in the indicated squares. Data are representative of five independent experiments. (C) Absolute numbers of MZ and FO B cells. Numbers were calculated by multiplying the fraction of each FCM B cell subset analyzed in (A) by the total number of CD19 + cells. Data are mean±S.D. obtained from five independent experiments. (D) The ratio of MZ to FO cell number was calculated. (E) Immunofluorescent staining of splenic cryosections with Cy3-anti-IgM and FITC-anti–MOMA-1. MOMA-1 + metalophilic macrophages define the border between the MZ B and FO B cells. Data are representative of five independent experiments. Scale bar, 100 µm. (F) Southern blot analysis of the DNA from the BM and spleen of the recipient mice. 4 Kb fragment represents wild-type, and 3 Kb fragment represent deletion.

Article Snippet: After air-drying, sections were fixed in 4% paraformaldehyde for 20 min. Immunohistochemical staining was done with Cy3-anti–mouse IgM and FITC-anti–MOMA-1 (Serotec) antibodies for 1 h at room temperature.

Techniques: Transplantation Assay, Staining, Southern Blot

Journal: PLoS ONE

Article Title: Dose-Dependent Induction of Murine Th1/Th2 Responses to Sheep Red Blood Cells Occurs in Two Steps: Antigen Presentation during Second Encounter Is Decisive

doi: 10.1371/journal.pone.0067746

Figure Lengend Snippet: Mice were primed with a LD or HD of SRBC. SRBC were labeled with CFSE before injection (green) and spleens were removed 1 h, 9 h and 24 h after LD and HD priming. Histological sections were prepared and the MZ were visualized by staining with Alexa 555-labelled antibodies against Moma-1, which is specific for metallophilic macrophages (red) (n = 3, one typical section is shown) ( A ). The MZ was isolated by laser-microdissection at indicated time points and mRNA expression of Il10 as marker for activated MZ B cells was analyzed. Each circle represents one animal (n = 6) ( B ). Significant differences in the expression of Il10 between primed mice compared to the controls are shown (**p<0.01, Mann-Whitney-U-test, n = 6). Mice were primed with a HD of SRBC. One footpad was challenged with SRBC 5 days after priming. The footpad thickness was measured between 1 and 4 days after challenge of wild type and LTβR−/− mice ( C ). Data are means ± SD. *indicate significant differences in footpad thickness compared to control mice (**p<0.01, Mann-Whitney-U-test, n = 6). Abbreviations: HD, high dose (10 9 ) ; LD, low dose (10 5 ).

Article Snippet: 10 µm thick cryosections were cut, stained with Biotin conjugated rat anti-mouse metallophilic macrophage monoclonal antibody (MOMA-1) (Acris Antibodies GmbH, Hiddenhausen) and visualized with Streptavidin Alexa Fluor® 555 (SAV-555; Invitrogen, Life Technologies, Germany).

Techniques: Labeling, Injection, Staining, Isolation, Laser Capture Microdissection, Expressing, Marker, MANN-WHITNEY

Journal: BMC Immunology

Article Title: Accumulation of marginal zone B cells and accelerated loss of follicular dendritic cells in NF-κB p50-deficient mice

doi: 10.1186/1471-2172-6-8

Figure Lengend Snippet: p50 -/- mice contain reduced numbers of MZ B cells that bind high levels of IgM-IC A. B cell subsets and IgM-IC binding. Splenocytes from B6 or p50 -/- mice were analyzed for B cell subsets by FACS using CD21/CD23 or B220/CD1d expression. Histograms on the right show binding of IgM-IC to MZ and FO B cells 1 h after i.v. injection (shaded region); clear profiles represent uninjected control mice. Although spleen cells from p50 -/- mice contained reduced numbers of MZ B cells, the cells present in the MZ B cell gate bind IgM-IC at high levels over background, similar to levels of binding of IgM-IC to B6 MZ B cells. B. In vitro binding assay showing binding of IgM-IC to MZ B cells (shaded region) compared with FO B cells (clear region). MZ B cells from both B6 and p50 -/- mice bind high levels of IgM-IC compared to FO B cells. C. Immunofluorescence shows the presence of IgM hi cells (red) in the MZ as demarcated by MOMA-1 (marginal metallophilic macrophages, blue) staining. p50 -/- mice possess some IgM + cells in the MZ outside of the MOMA-1 ring, consistent with the presence of appropriately positioned MZ B cells in these mice.

Article Snippet: Antibodies utilized included anti-MOMA-1 (1/25; Serotec, Raleigh NC) visualized with a goat anti-rat IgG-AMCA (1/100; Jackson Immunoresearch), anti-IgM-Texas Red (1/100; BD Biosciences), and anti-laminin (1/500; Sigma) visualized with an anti-rabbit IgG-FITC (1/100; Jackson Immunoresearch).

Techniques: Binding Assay, Expressing, Injection, In Vitro, Immunofluorescence, Staining