Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Resident immune cells of the avascular lens: Mediators of the injury and fibrotic response of the lens

doi: 10.1096/fj.202002200R

Figure Lengend Snippet: Resident immune cells are present among the human, mouse, and chicken lens epithelium. CD45 and β2 integrin+ resident immune cells (green) are found localized among the epithelial cells (indicated by double arrows) in explants from (A-C) human, (D-F) mouse, and (G-I) chick lens tissue. Lens tissue was counterstained with DAPI to identify nuclei (blue). Within the lens tissue from each species, many of these CD45+ and β2 integrin+ immune cells have dendritic morphologies indicated by the arrowheads. The dendritic-like processes of resident immune cells indicated by asterisks in A, D, G are highlighted by 3D surface structures created by Imaris software from confocal z-stack images and shown in (B, C, E, F, H, and I). In 3D structure images, the dendritic processes of immune cells are seen reaching between the lens epithelial cells. Magnification bars = 20 μm. Studies are representative of at least three individual lenses

Article Snippet: Primary antibodies for the chick lens study used included: MHC-II (Bio-Rad, Hercules, CA, USA), CD45 (BioRad, Hercules, CA, USA ), CD14 (BioRad, Hercules, CA, USA), Vimentin (Abcam, Cambridge, MA, USA) Vimentin (gift, P. Fitzgerald, University of California, Davis, CA), β2-integrin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and CD45 (Abcam, Cambridge, MA, USA).

Techniques: Software

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: Resident immune cells of the avascular lens: Mediators of the injury and fibrotic response of the lens

doi: 10.1096/fj.202002200R

Figure Lengend Snippet: Resident immune cells migrate to the wound edge in response to mock cataract surgery wounding. A-H, Ex vivo mock cataract surgery chick explants 1 hour postinjury were labeled for the leukocyte specific integrin β2 (CD18) (green, A, C, E, and F) and co-labeled for the leukocyte antigen CD45 (red, B), vimentin (red, D) or the monocyte/macrophage lineage marker CD14 (red G, H). Explants were labeled for F-actin (white, A-H) to delineate the lens epithelial cells. CD45/β2+ cells were also counterstained with DAPI to identify nuclei (A,B). β2 integrin/CD45+ resident leukocytes migrated to the wound edge in response to mock cataract surgery. β2 integrin+ immune cells at the wound edge where enriched for vimentin (C,D) and subpopulations of β2 integrin+ cells expressed CD14 (F,G), identifying that subsets of resident immune cells are from the monocyte/macrophage lineage. E,H, 3D surface images from z-stack confocal images of images presented as single optical planes in C and G highlight the morphology of the resident immune cells extending protrusions at the wound edge. Asterisks in (E,H) indicate the position of the same cells shown in (C,G), respectively. Magnification bars = 20 μm. Studies are representative of at least three independent experiments

Article Snippet: Primary antibodies for the chick lens study used included: MHC-II (Bio-Rad, Hercules, CA, USA), CD45 (BioRad, Hercules, CA, USA ), CD14 (BioRad, Hercules, CA, USA), Vimentin (Abcam, Cambridge, MA, USA) Vimentin (gift, P. Fitzgerald, University of California, Davis, CA), β2-integrin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and CD45 (Abcam, Cambridge, MA, USA).

Techniques: Ex Vivo, Labeling, Marker

Journal: Molecular Biology of the Cell

Article Title: Ligand-specific binding forces of LFA-1 and Mac-1 in neutrophil adhesion and crawling

doi: 10.1091/mbc.E16-12-0827

Figure Lengend Snippet: AFM tests for β2 integrin-ligand bonds. (A) Schematic of AFM setup. A PZT was used to drive the movement of an AFM cantilever. Adhesion events and forced bond rupture signals were collected from a QPD that measures the deflection of a laser beam reflected on the cantilever. (B) AFM functionalization. Recombinant human LFA-1s or Mac-1s were adsorbed onto the AFM tip and treated with TS1/18 mAbs or Mn 2+ to obtain low- or high-affinity conformation integrins, respectively. Soluble ligand (ICAM-1, ICAM-2, RAGE, JAM-A, or JAM-C)–IgG Fc chimeras were coated via anti-IgG Fc secondary antibodies precoated on a Petri dish. (C) Typical force–displacement curves. An LFA-1– or Mac-1–captured AFM tip was driven to approach to (from left to right, blue lines), contact, and retract from (from right to left, red lines) a ligand-coated Petri dish. Adhesion was visualized from cantilever deflection and rupture force ( f r ) was measured from the force–displacement curve (middle and lower trajectories). k s is the system spring constant derived from the slope of the adhesive event. (D, E) Binding specificity. The LFA-1– or Mac-1–captured tip was pretreated with isotype control (closed bars) or LFA-1/Mac-1 blocking (open bars) mAbs. Adhesion probabilities were measured between the tip and various ligand-coated Petri dishes with TS1/18 (D) or Mn 2+ (E). All measurements were acquired at a cantilever approach and retraction velocity of 1 μm/s, a contact duration of 50 ms, and a compression force of 200 pN. An anti-IgG Fc secondary antibody–coated substrate was used as a negative control. Data are presented as the mean ± SEM of three or four tips in each case. Significant differences are indicated by *, p < 0.05; **, p < 0.01; ***, p < 0.001 between each ligand and negative control, by # , p < 0.05; ## , p < 0.01; ### , p < 0.001 between each blocking group and isotype control, or by $ , p < 0.05; $$ , p < 0.01; $$$ , p < 0.001 among various ligands. (F) Typical rupture force distributions for the interactions between LFA-1 and ICAM-1 with TS1/18 at the indicated loading rates. Total 251–394 single-bond rupture force data at each loading rate were collected and analyzed using a force bin of 50 pN.

Article Snippet: For example, shear stress regulates leukocyte adhesion , induces activation and cleavage of β2 integrins during PMN migration ( Makino et al. , 2007 ), triggers guanine nucleotide exchange factor (GEF)-H1–dependent spreading and crawling of PMNs ( Fine et al. , 2016 ), and promotes lymphocyte transmigration ( Cinamon et al. , 2001 ).

Techniques: Recombinant, Derivative Assay, Adhesive, Binding Assay, Control, Blocking Assay, Negative Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Entamoeba histolytica Induce Signaling via Raf/MEK/ERK for Neutrophil Extracellular Trap (NET) Formation

doi: 10.3389/fcimb.2018.00226

Figure Lengend Snippet: Entamoeba histolytica- induced NET formation is independent on Syk, TAK1, β2 integrins, PI3K, and p38 MAPK. Human neutrophils were left untreated (—), stimulated with 20 nM phorbol 12-myristate 13-acetate (PMA), or with E. histolytica trophozoites. PMN were previously treated with solvent alone (—) or 1 μM iSyk, a Syk inhibitor, or 10 μg/ml of the blocking monoclonal antibody anti-β2 integrins (IB4), or 10 nM LLZ 1640-2 (LLZ), a TAK1 inhibitor, or 50 nM Wortmannin (Wort), a PI3K inhibitor, or 200 nM SB203580 (SB), a p38 MAPK inhibitor. The relative amount of NETs was estimated from SYTOX® Green fluorescence in relative fluorescent units (RFU) at 4 h after stimulation. Data are mean ± SEM of three experiments. Asterisk denotes a condition that was statistically different from control ( p < 0.05).

Article Snippet: The involvement of β2 integrins in NET formation might be more related to the adhesion requirement of neutrophils to form NETs (Brinkmann et al., ) than to a signaling capacity of the integrin.

Techniques: Solvent, Blocking Assay, Fluorescence, Control

Journal: eBioMedicine

Article Title: Cis -interaction between CD52 and T cell receptor complex interferes with CD4 + T cell activation in acute decompensation of cirrhosis

doi: 10.1016/j.ebiom.2024.105336

Figure Lengend Snippet: Membrane proteins identified by proximity labelling.

Article Snippet: Recombinant human TCRβ1, CD3ε, CD3γ, CD43, CD44, CD48 (Cusabio Biotech), CD4 (ABclonal, Port Talbot, UK), integrin β2 (Assay Genie, Dublin, Ireland) proteins, recombinant human CD52 monoclonal antibody, Siglec-10, and IgG1 proteins (R&D Systems) were labelled with 4-((4-(dimethylamino)phenyl)azo)benzoic acid (DABCYL) using DABCYL-succinimidyl ester.

Techniques: Membrane

Journal: Oncotarget

Article Title: Bone marrow stroma-induced resistance of chronic lymphocytic leukemia cells to arsenic trioxide involves Mcl-1 upregulation and is overcome by inhibiting the PI3Kδ or PKCβ signaling pathways

doi:

Figure Lengend Snippet: Clinical characteristics of the CLL patients

Article Snippet: RpAb to Bim (559685) and mAb to β2 integrin (556084) were from BD Pharmingen (Franklin Lakes, NJ). mAbs to vinculin (V9131) and β-actin (A5316) were from Sigma-Aldrich (St. Louis, MO). mAb to CD19 was from Diaclone (Besançon, France). mAbs against CD38 (16BDH), CD3 (T3B), α4 integrin (HP2/1, function blocking), α4 integrin (HP1/7, inactive control, isotype matched for HP2/1 and anti-β2 Abs), and β1 integrin (Alex1/4) were from Dr. F. Sánchez-Madrid (Hospital de la Princesa, Madrid, Spain).

Techniques: