α ly6g clone 1a8

For anti-Gr-1-mediated depletions, mice were injected i.p. with 250μg of either α-Gr-1 (clone RB6-8C5) or rat IgG2b isotype control antibody (clone LTF-2) (Bio X cell). Mice were injected 16 hours before infection and then again 48 hours post-infection. For anti-Ly6G-mediated depletions, mice were injected i.p. with 250μg of either α-Ly6G (clone 1A8) or rat IgG2a isotype control antibody (clone 2A3) (Bio X Cell). Mice were injected 48 hours prior to infection and then again every 24 hours until the completion of the infection. 24 or 72 hours post-infection, lungs were harvested and CFUs were enumerated. The efficiency of neutrophil or monocyte depletion was monitored by flow cytometry.

The function of neutrophils during acute infections by S. pyogenes were examined in B6_HLA mice between 8–12 weeks old. Neutrophils were depleted in vivo by intraperitoneally injecting mice with 250 μg mAb αLy6G clone 1A8 (BioXcell, NH, USA) 24 hours before and 24 hours following nasopharyngeal and skin infections. Control mice received rat IgG2a clone 2A3 (BioXCell, NH, USA). Depletion of circulating neutrophils has been confirmed in previous studies using flow cytometry of Ly6G⁺ expressing populations in blood [47 (link),74 (link),75 (link)]. Bacterial burden in nasal turbinates was examined at both 24 and 48 hours following infection.