Anti ter119 ter 119

Mice were sacrificed after 60 days and spleens, inguinal and mesenteric lymph nodes were harvested and homogenized into single cell suspensions using HBSS + 2% FBS (Gibco; Gaithersburg, MD). Spleen cells were further treated with ammonium chloride lysing solution to remove erythrocytes. The following anti-mouse antibodies were used for immunophenotyping: anti-CD1d (1D1) (BD Pharmingen; San Diego, CA), anti-CD3 (145-2C11) (BD Pharmingen), anti-CD4 (GK1.5) (eBioscience, San Diego, CA), anti-CD5 (53–7.3) (BD Pharmingen), anti-CD8 (53–6.7) (eBioscience), anti-CD11b (M1/70) (eBioscience), anti-CD19 (6D5) (Caltag Laboratories; Burlingame, CA), anti-CD25 (PC61) (BD Pharmingen), anti-Ter119 (TER-119) (eBioscience), anti-B220 (RA3-6B5) (BD Pharmingen), anti-DX5 (DX5) (eBioscience), anti-Gr-1 (RB6-8C5) (eBioscience), and anti-FoxP3 (FJK-16s) (eBioscience). Staining for FoxP3 was performed according to the manufacturer’s protocol (eBioscience). All samples were acquired on a FACSCalibur flow cytometer (BD Biosciences; San Jose, CA) and analyzed by FlowJo software (Ashland, OR).

Cells from MLNs, spleen, blood, and bone marrow were stained for live dead (Zombie UV, Biolegend) and Fc block (eBiosciences) prior to cell surface cellular markers staining. Samples were read on a BD LSR Fortessa flow cytometer (BD Biosciences), and data were analyzed using FlowJo X (Tree Star, Inc).
Cell surface markers: anti-B220 (RA3-6B2), anti-CD19 (6D5), anti-CD3ε (17A2), anti-CD4 (RM4.5), anti-CD8α (53-6.7), anti-CD279 (PD-1) (29F.1A12), anti-CD185/CXCR5 (L138D7), anti-CD93 (AA4.1), anti-CD25 (3C7), and anti-CD117/c-kit (2B8) were purchased from Biolegend. Anti-CD43 (S11), anti-CD103 (2E7), anti-CD11c (N418), anti-CD317/PDCA-1 (927), anti CD11b (M1/70), anti-CD64 (X54-5/7.1), anti-I-A/I-E (M5/114.15.2), and anti-CD23 (B3B4) were purchased from BD Biosciences. Anti-CD45 (30-F11), anti-ly6G (RB6-8C5), anti-NK.1 (PK136), and anti-Ter119 (Ter-119) were purchased from eBiosciences.