Depleting Immune Cells and Neutralizing Cytokines in Mice

To deplete immune cells and neutralize cytokines, mice were injected intraperitoneally with each antibody as previously described [57 (link)] with some modifications. To deplete neutrophils during early infection, mice infected with M. intracellulare were intraperitoneally injected with an initial dose of 400 μg followed by 300 μg of anti-Ly6G antibody (clone 1A8, BioXcell) or an initial dose of 400 μg followed by 300 μg of rat IgG2a isotype control (clone 2A3, BioXcell) every other day from 1 to 11 days postinfection (Fig 5A). To deplete neutrophils during chronic infection, mice infected with M. intracellulare were intraperitoneally injected with an initial dose of 400 μg followed by 300 μg of anti-Ly6G antibody (clone 1A8, BioXcell) or an initial dose of 400 μg followed by 300 μg of rat IgG2a isotype control (clone 2A3, BioXcell) 3 times per week for 2 weeks from 32 to 34 weeks postinfection (Fig 5I). To neutralize IL-17A, mice infected with M. intracellulare were intraperitoneally injected with 100 μg of anti-IL-17A antibody (clone 17F3, BioXcell) or 100 μg of mouse IgG1 isotype control (clone MOPC-21, BioXcell) every other day from 1 to 11 days postinfection.