Anti cd3e 145 2c11

Mice were injected i.p. with a mix of 100 μg anti-CD8 [TIB105(53.6.72)] and 100 μg anti-CD4 (GK1.5), or isotype control on day −3, 0, and 3 relative to viral challenge. Spleens were harvested from mice receiving the same injections 3 days after injection and evaluated by flow cytometry for depletion efficacy using anti-CD3e (145-2C11; Tonbo Biosciences), anti-CD45R (RA3-6B2; Tonbo Biosciences), anti-CD4 (1540-11; Southern Biotech) and anti-CD8 (553033; BD Biosciences) and isotype controls.

DCs from spleens of BALB/c mice were prepared using the GentleMACS dissociator (Miltenyi Biotech) according to the manufacturer's protocol. Briefly, spleens were dissociated in GentleMACS C tubes in medium containing collagenase and DNase, incubated for 15 min at 37 C before adding EDTA at a final concentration of 10 mM. Erythrocytes were lysed by incubation with ACT buffer for 5 min on ice. Finally, cells were filtered through a 75 mm Nylon cell strainer. The following Abs were used for subsequent flow cytometry analysis: anti-CD3e (145-2C11; Tonbo Bio- sciences), anti-CD19 (1D3; Tonbo Biosciences), anti-CD49b (DX5; eBioscience), anti-Ly6G (1A8), anti-CD45R (RA3-6B2; Tonbo Biosciences), anti-MHC-II (M5/114.15.2; BioLegend), anti-CD11c (N418; Tonbo Bio- sciences), anti-CD11b (M1/70; Tonbo Biosciences), and anti-CD24 (M1/69; BioLegend).