Anti ly6g antibody clone 1a8

CD73^−/− mice were intraperitoneally administered 300 μg of anti‐Ly6G antibody (clone1A8, BioXCell, West Lebanon, NH) at Day ‐2, 0, and 1 day after the last caerulein injection. IgG2a isotype (clone 2A3, BioXCell, West Lebanon, NH) was used for control. Mice were euthanized at Day 4.

C57BL/6J mice were obtained from Charles River (Sulzfeld, Bavaria, Germany). Animals were maintained under pathogen-free conditions in the research animal facility of Tuebingen University, Tuebingen, Germany (K08/19G). All experimental animal procedures were conducted according to the guidelines from Directive 2010/63/EU of the European Parliament on the protection of animals used for scientific purposes and the German federal and state regulations.
Neutrophils were systemically depleted in newborn mice by administering a depleting Ly6G antibody to pregnant and subsequently to lactating dams. For this purpose, wild-type mice (C57BL/6J) were terminally mated. Two days before the expected litter date, the dam received an intravenous (IV) injection of a depleting anti-Ly6G antibody (clone 1A8, BioXCell, Lebanon, NH, USA) or an isotype control antibody (IgG2A, BioXCell, Lebanon, NH, USA) into the tail vein. This achieved depletion of all peripheral neutrophils. Because reconstitution of cells from the bone marrow occurs within three days [42 (link)], the injection was repeated every three days until the offspring were sacrificed. The first injection after birth was again given IV into the tail vein of the dam, and all subsequent injections were given intraperitoneally (IP).

Mice were treated with anti-Ly6G antibody (clone 1A8; Bio-X-Cell, West Lebanon, NH) at 200 ug per mouse intraperitoneally 3X weekly for 3–4 weeks starting 3 days before infection with H. hepaticus. Treatment with antibody continued concurrent with H. hepaticus infection for a duration of 3 weeks, with euthanasia occurring at age 15 weeks. Treated mice were then compared with mice receiving a comparable dose of sham isotype antibody alone. Depletion of neutrophils was confirmed by undetectably low levels of MPO+ cells in spleens of mice treated with anti-Ly-6G antibody compared to sham-treated controls.

Mice were administered an intraperitoneal dose of 0.4 mg of anti-Ly6G antibody (clone 1A8, BioXCell) or a rat IgG2a isotype control (clone 2A3, BioXCell). The antibody was injected on a schedule of four time points: one day before ligature placement, and 2, 5, and 8 days after ligature placement. Bone loss was subsequently measured 10 days after ligature placement.

Depletion of CD8⁺ T cells in a lung metastasis model using LLC1.1 cells was performed with 10 μg anti-CD8 antibody (clone 2.43; BioXCell) i.v. injected one day prior to primary tumor removal. Mice received another 15 μg of the antibody by i.v. injection two days after the tumor removal. Control mice received the isotype control antibody (clone 2A3, BioXCell) in parallel. In the liver metastasis model, mice received 10 μg Ab injection 1 day before tumor cell injection and the second treatment with 15 μg Ab two days after tumor cell injection.
Neutrophil depletion was achieved by intraperitoneal injection (i.p.) of anti-Ly6G antibody (clone 1A8; BioXCell). Initial injection of 500 μg/mouse was performed 24 hours prior to intrasplenic injection of MC-38GFP cells. Mice were treated three times weekly with 100 μg i.p. until the termination at day 27. Control animals were injected with isotype-control Ab (clone 2A3, BioXCell) in parallel.