Journal: iScience

Article Title: Preclinical safety and efficacy of a therapeutic antibody that targets SARS-CoV-2 at the sotrovimab face but is escaped by Omicron

doi: 10.1016/j.isci.2023.106323

Figure Lengend Snippet:

Article Snippet: Anti-light-kappa-PE (clone IS11-24D5) , Miltenyi Biotec , Cat# 130-123-246; RRID: AB_2802012.

Techniques: Virus, Recombinant, Blocking Assay, Saline, Enzyme-linked Immunosorbent Assay, Membrane, Plasmid Preparation, Expressing, Software

Journal: Acta neuropathologica communications

Article Title: Molecular signatures of inherited and acquired sporadic late onset nemaline myopathies.

doi: 10.1186/s40478-023-01518-9

Figure Lengend Snippet: Fig. 4 Analysis of immunoglobulin expression and autoimmune markers in inherited and sporadic late-onset nemaline myopathies (A) Proteomic analysis showed an increased abundance of IgG heavy chains and both kappa and lambda light chains in nemaline rod areas in sporadic late-onset nemaline myopathy (SLONM) relative to inherited nemaline myopathies (iNM). There was also a trend towards an increased abundance of immunoglobulin chains in the nemaline rod areas of SLONM relative to rod-free areas. Striped bars indicate immunoglobulin chains that were only detected in one of the groups of samples compared *p < 0.05. (B) Immunofluorescent staining demonstrated accumulation of kappa light chains in atrophic fibers in 38% of SLONM biopsies (bottom row), but not in iNM (top row). Scale bar = 400 µm. (C) Immunoglobulin genes were also differentially expressed between SLONM and iNM. (D) Immunostaining for Major Histocompatibility Antigen-I (MHC-I) showed reactivity in SLONM biopsies, occurring predominantly in atrophic fibers (first panel, SLONM with monoclonal gammopathy; second panel, SLONM without monoclonal gammopathy). Biopsies from patients with inherited nemaline myopathy showed no MHC-I reactivity or occasional fibers faintly reactive for MHC-I (third panel, adult patient with ACTA1 nemaline myopathy). MHC-I reactivity was absent in healthy control muscle (fourth panel). Scale bar = 100 µm

Article Snippet: Frozen skeletal muscle sections (8 μm) were post-fixed in 4% paraformaldehyde for 5 min at room temperature prior to immunostaining with laminin (L0663, Sigma, dilution 1:250) and human kappa light chain (NBP269235, Novus Biologicals, Centennial, CO, dilution 1:200) antibodies as previously described [18].

Techniques: Expressing, Staining, Immunostaining, Control

Journal: Mucosal immunology

Article Title: Interleukin (IL)-4, IL-13, and IL-17A differentially affect the profibrotic and proinflammatory functions of fibrocytes from asthmatic patients.

doi: 10.1038/mi.2011.60

Figure Lengend Snippet: Figure 1 Comparison of the frequency of CD34 + COL1 + ( a ) and CD34 + COL1A1 mRNA + fibrocytes ( b ) in the peripheral blood of healthy controls and asthmatic patients. The percentage of cells coexpressing CD34 and COL1 or CD34 and COL1A1 mRNA was calculated on cytospins of total peripheral blood leukocytes. The horizontal lines indicate the medians. * * * P < 0.001 vs. healthy controls by the Mann – Whitney test.

Article Snippet: For intracellular staining of COL1, cells were permeabilized with 0.3 % saponin solution (Sigma-Aldrich, St Louis, MO) in Tris-buffered saline and sequentially 148 VOLUME 5 NUMBER 2 | MARCH 2012 | www.nature.com/mi incubated with a rabbit anti-human COL1 ( 2 chain) (Novus Biologicals, Littletone, CO), a biotinylated goat anti-rabbit secondary antibody (Novus Biologicals), and rhodamine red X-conjugated streptavidin (Jackson ImmunoResearch, Bar Harbor, ME).

Techniques: Comparison, MANN-WHITNEY

Journal: Mucosal immunology

Article Title: Interleukin (IL)-4, IL-13, and IL-17A differentially affect the profibrotic and proinflammatory functions of fibrocytes from asthmatic patients.

doi: 10.1038/mi.2011.60

Figure Lengend Snippet: Figure 2 Isolation and phenotypic analysis of circulating fibrocytes. CD34 + cells were isolated from the peripheral blood by positive immunomagnetic selection and cultured for 5 days to obtain a pure population of mature fibrocytes. The non-adherent cells were removed after the first 48 h and the adherent cells were reincubated for the next 3 days in fresh medium. Flow cytometric analysis of the isolated cells was performed soon after the immunomagnetic selection ( a ) and on day 5 ( c ). The horizontal and vertical lines in the representative dot plots mark fluorescence intensity greater than that observed with isotype-matched controls for the anti-CD45, anti-CD34, and anti-COL1 antibodies. The morphology of the cells in culture was monitored under an inverted microscope and the microphotographs in panel b show representative images in phase-contrast mode taken at the indicated points in time. The expression of the type I interleukin (IL)-4 receptor subunits (IL-4R / common -chain ( c)), type II IL-4 receptor subunits (IL-4R / IL-13R 1), IL-13R 2, and IL-17RA on the membrane of control and asthmatic fibrocytes were evaluated by flow cytometry on day 5 ( d ). The relative level of expression was quantified by subtracting the median fluorescence intensity (MFI) of cells stained with each specific antibody from the MFI of cells stained with the corresponding isotype control and dividing the obtained value by the MFI of unstained cells. Data are expressed as the means and s.d. * P < 0.05 vs. control fibrocytes; * * P < 0.01 vs. control fibrocytes by the unpaired Student ’ s t -test; n = 5.

Article Snippet: For intracellular staining of COL1, cells were permeabilized with 0.3 % saponin solution (Sigma-Aldrich, St Louis, MO) in Tris-buffered saline and sequentially 148 VOLUME 5 NUMBER 2 | MARCH 2012 | www.nature.com/mi incubated with a rabbit anti-human COL1 ( 2 chain) (Novus Biologicals, Littletone, CO), a biotinylated goat anti-rabbit secondary antibody (Novus Biologicals), and rhodamine red X-conjugated streptavidin (Jackson ImmunoResearch, Bar Harbor, ME).

Techniques: Isolation, Selection, Cell Culture, Fluorescence, Inverted Microscopy, Expressing, Membrane, Control, Flow Cytometry, Staining