anti-ly6g antibody clone 1a8  (Bio X Cell)

Journal: Frontiers in Immunology

Article Title: Patrolling monocytes mediate virus neutralizing IgG effector functions: beyond neutralization capacity

doi: 10.3389/fimmu.2025.1600056

Figure Lengend Snippet: Macrophages and inflammatory monocytes are dispensable for nAbs-mediated LCMV control. (A) Flow cytometric analysis of monocytes subsets, CD45 + Lin - (CD45R + CD8 + CD4 + NK + ), CD11b + Ly6G - , in the peripheral blood of MC21 or isotype (rat IgG2a)-treated mice 3 days pot infection (d.p.i). Mice were infected with LCMV-WE (2x10 5 PFU) on day 0, preceded by treatment with 25 µg MC21 or isotype (rat IgG2b) on day -1 and again on day 1. See ( Supplementary Figure S2 ) for gating strategy of the monocyte subsets. (B) Numbers of patrolling monocytes (-Mo), inflammatory monocytes (-Mo), neutrophils, and lymphocytes in the peripheral blood of MC21 or isotype (rat IgG2a)-treated mice 3 d.p.i. Results show the pooled data from two independent experiments with similar results. (n=3–5 mice/group/experiment). (C) Virus titres (3 d.p.i) in spleen of WT mice infected with LCMV-WE (2x10 5 PFU) on day 0 and treated on day 1 with Wen3 (350 µg) or isotype (IgG2a), On day -1 and day 1 mice were treated with 25 µg MC21 or isotype (rat IgG2b). Results show the pooled data from two independent experiments with similar results. (n=3–5 mice/group/experiment). (D) Schematic presentation of the experiments in (E-G) . (E) Numbers of patrolling monocytes (-Mo), inflammatory monocytes (-Mo), neutrophils, and lymphocytes in the peripheral blood and spleen (3 d.p.i.) of mice treated with 10 µL/g bodyweight control or clodronate liposomes one day before LCMV infection (2x10 5 PFU). Results show the pooled data from two independent experiments with similar results. Experiment was repeated three times with similar results (n=4–5 mice/group/experiment). (F) Shown are the virus titres (3 d.p.i.) in spleen of WT mice infected with LCMV-WE (2x10 5 PFU) on day 0 and treated on day 1 with Wen3 (350 µg) or isotype (IgG2a). On day -1 mice were treated with 10µL/g bodyweight control or clodronate liposomes. Results show the pooled data from two independent experiments with similar results. Experiment was repeated three time with similar results (n=3–5 mice/group/experiment). (G) Spleen sections collected 3 d.p.i. were stained for F4/80 (blue), CD169 (green), and LCMV nucleoprotein (−NP) (red). Scale bar 300 µm. Shown are representative pictures from three independent experiments (n=2–4 mice/group/experiment). Statistical analysis was performed by using the One-way ANOVA test (C, F) or Student’s t-test (B, E) . *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.001, ns, non-significant. Horizontal dotted lines indicating the detection limit.

Article Snippet: Depletion antibodies utilized were CD115 (Clone: AFS98), Ly6G (Clone: 1A8), NK (Clone: PK136), all procured from BioXcell, and each was administered at a dosage of 200 μg per mouse (except CD115 100 μg).

Techniques: Control, Infection, Virus, Liposomes, Staining

Journal: Frontiers in Immunology

Article Title: Patrolling monocytes mediate virus neutralizing IgG effector functions: beyond neutralization capacity

doi: 10.3389/fimmu.2025.1600056

Figure Lengend Snippet: Contribution of patrolling monocytes for antiviral activity of Wen3. (A) Schematic presentation of the experiments in (B-E) . (B) Flow cytometric analysis of monocyte subsets in the peripheral blood and spleen of control or clodronate liposomes-treated mice 3 days post infection (d.p.i.). Animals were treated with 0.5 µL/g control or clodronate liposomes (Clo. (low) ) on day -1 and 1. Mice were infected with LCMV-WE (2x10 5 PFU) on day 0. See ( Supplementary Figure S2 ) for gating strategy of the monocyte subsets. (C) Numbers of patrolling monocytes (-Mo), inflammatory monocytes (-Mo), neutrophils, and lymphocytes/macrophages in the peripheral blood and spleen of control or clodronate liposome-treated mice 3 d.p.i. Animals were treated with 0.5µL/g control or clodronate liposomes (Clo. (low) ) on day -1 and 1. mice were infected with LCMV-WE (2x10 5 PFU) on day 0. Results show the pooled data from two independent experiments with similar results. (n=3–5 mice/group/experiment). (D) Virus titres (3 d.p.i.) in spleen of WT mice infected with LCMV-WE (2x10 5 PFU) on day 0 and treated on day 1 with Wen3 (350 µg) or isotype (IgG2a), On day -1 and day 1 mice were treated with 0.5µL/g bodyweight control or clodronate liposomes (Clo. (low) ). Results show the pooled data from three independent experiments with similar results (n=3–4 mice/group/experiment). (E) Spleen sections collected 3 d.p.i. were stained for CD169 (green) and LCMV nucleoprotein (−NP) (red). Shown are the representative merged images from three independent experiments. Scale bar= 300 µm. (F) Virus titres (3 d.p.i.) in spleen of WT mice infected with LCMV-WE (2x10 5 PFU) on day 0 and treated on day 1 with neutralizing serum (Neutralizing S.) or naïve serum (see methods). On day -1 and day 1 mice were treated with 0.5µL/g control or clodronate liposomes (Clo. (low) ). Results show the pooled data from two independent experiments with similar results (n=3–5 mice/group/experiment). (G) Numbers of patrolling monocytes (-Mo), inflammatory monocytes (-Mo), neutrophils, along with lymphocytes in the peripheral blood or macrophages in spleen (3 d.p.i.) of mice treated with anti-CD115 or Isotype. Mice were infected with LCMV-WE (2x10 5 PFU) on day 0, preceded by treatment with 100 µg anti-CD115 or isotype (rat IgG2b) on day -1 and again on day 1. Results show the pooled data from two independent experiments with similar results (n=3 mice/group/experiment). (H) Shown are the virus titres (3 d.p.i.) in spleen of WT mice infected with LCMV-WE (2x10 5 PFU) on day 0 and treated on day 1 with Wen3 (350 µg) or isotype (IgG2a). On day -1 and day 1 mice were treated with 100 µg anti-CD115 or isotype. Results show the pooled data from two independent experiments with similar results (n=3 mice/group/experiment). (I) Shown are the virus titres (3 d.p.i.) in spleen of CD11c-cre +/+ /iDTR and CD11c-cre tg/+ /iDTR mice infected with LCMV-WE (2x10 5 PFU) on day 0 and treated on day 1 with Wen3 (350 µg) or isotype (IgG2a), on day -1 and day 1 mice were treated with diphtheria toxin 12ng/g bodyweight. Results show the pooled data from two independent experiments with similar results (n=3–4 mice/group/experiment). (J) Shown are the virus titres (3 d.p.i.) in spleen of WT mice infected with LCMV-WE (2x10 5 PFU) on day 0 and treated on day 1 with Wen3 (350 µg) or isotype (IgG2a), mice were treated with 200 µg 9E9 or isotype for blocking FcγRIV. Results show the pooled data from three independent experiments with similar results (n=3–4 mice/group/experiment). (K) Heat map showing the expression of FcγRIV on CD11b + Ly6G - population gated from CD45 + Lin - (CD45R + CD8 + CD4 + ), 24 hour following infection with LCMV. Statistical analysis was performed by using the Student’s t-test (C, G) or One-way ANOVA test (D, F, H–J) . *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.001, ns, non-significant. Horizontal dotted lines indicating the detection limit.

Article Snippet: Depletion antibodies utilized were CD115 (Clone: AFS98), Ly6G (Clone: 1A8), NK (Clone: PK136), all procured from BioXcell, and each was administered at a dosage of 200 μg per mouse (except CD115 100 μg).

Techniques: Activity Assay, Control, Liposomes, Infection, Virus, Staining, Blocking Assay, Expressing

Journal: JCI Insight

Article Title: Combination treatment with anti-RANKL and antibiotics for preventing joint destruction in septic arthritis

doi: 10.1172/jci.insight.184954

Figure Lengend Snippet: ( A ) The protective effect of neutrophils was examined by depleting mice of neutrophils ( n = 10/group) using the anti-Ly6G antibody (Neutrophil depletion) or an isotype control (Controls). Knee swelling was measured in millimeters over a period of 7 days after intra-articular (i.a.) injection of 20 μL of PBS containing S . aureus LS-1 strain (4 × 10 3 CFU/knee) into the knee joints of NMRI mice. ( B ) Bacterial counts in the mouse knee joints ( n = 5/group) were assessed. ( C ) Bone erosion scores of the knee joints ( n = 5/group) were determined after micro-CT (μCT) scan. ( E ) Cumulative survival was monitored daily. ( F ) The pathogenic role of monocytes was investigated by depleting mice of monocytes ( n = 22/group) using clodronate liposomes (Monocyte depletion) or PBS control liposomes (Controls), and knee swelling was followed for 10 days after infection using the same strategy. ( G ) Bacterial counts in mouse knee joints in the monocyte depletion group ( n = 5) and controls ( n = 6). ( H ) Bone erosion scores for the monocyte depletion group ( n = 7) and controls ( n = 16) and ( I ) cumulative survival. ( D ) Representative μCT images of knee joints from the control, neutrophil depletion, and monocyte depletion groups. Arrows indicate bone erosion. The data were pooled from 2 independent experiments. Statistical evaluations were performed using the Mann-Whitney test ( A – C and F – H ) or log-rank (Mantel-Cox) test ( E and I ). Data are presented as mean with SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Selective depletion of murine blood neutrophils was accomplished using the anti-Ly6G monoclonal antibody (clone 1A8; BioXCell), as previously documented ( ).

Techniques: Control, Injection, Micro-CT, Liposomes, Infection, MANN-WHITNEY

Journal: JCI Insight

Article Title: Combination treatment with anti-RANKL and antibiotics for preventing joint destruction in septic arthritis

doi: 10.1172/jci.insight.184954

Figure Lengend Snippet: Knee joints from NMRI mice, collected on day 3 after intra-articular (i.a.) injection of 20 μL of PBS or Lpl1(+sp) (4 μg/knee) or PBS containing S . aureus LS-1 strain (4 × 10 3 CFU/knee), underwent anti-CD68 immunohistochemistry examination ( A – C ). Scale bar: 100 μm. Lpl1(+sp), S . aureus intact Lpp. Knee synovial tissues from mice that were injected i.a. with 20 μL of PBS ( n = 4) or PBS containing S . aureus LS-1 strain (4 × 10 3 CFU/knee, n = 6) were analyzed on day 3 after injection using flow cytometry. t-Distributed stochastic neighbor embedding (tSNE) analysis of monocyte subsets based on Ly6C expression with gating on CD11b + CD45 + Ly6G – population was conducted ( D ). Representative mean fluorescence intensity (MFI) histograms of monocyte subsets for RANK, c-Fms, and CCR2 are shown ( E ). Statistical analyses of RANK, c-Fms, and CCR2 levels are presented ( F ). Statistical evaluations were performed using 1-way ANOVA with Holm-Šídák multiple-comparison test, with data presented as mean with SEM. * P < 0.05; ** P < 0.01; **** P < 0.0001.

Article Snippet: Selective depletion of murine blood neutrophils was accomplished using the anti-Ly6G monoclonal antibody (clone 1A8; BioXCell), as previously documented ( ).

Techniques: Injection, Immunohistochemistry, Flow Cytometry, Expressing, Fluorescence, Comparison