Anti cd19 1d3

Spleens were collected and single cell suspensions made. For T helper cell analysis, cells were surfaced stained with anti-CD4 (RM 4-5; eBioscience), permiabalized and intracellular stained with IFN-γ (XMG1.2; eBioscience), IL-4 (BVD6-24G2; eBioscience), RORγ (B2D; eBioscience), CD25 (PC61.5; eBioscience) or FoxP3 (FJK-16s; eBioscience). Representative gating in Supplemental Data Figure S1. For CD8⁺ T cells, cell surfaces were stained with anti- CD8a (53-6.7; eBioscience) and anti- CD3e (145-2C11; eBioscience). For B cells, cell surfaces were stained with anti- CD21/CD35 (8D9; eBioscience) and anti- CD19 (1D3; eBioscience). T cell activation markers CD25 (eBio3C7; eBioscience) and CD69 (H1.2F3, Biolegend) were measured using flow cytometry.
For CBC exams ≈ 50 μL of blood was collected into EDTA coated tubes via retro-orbital bleed. VetScan HM5 Analyzer was used.

Cellular suspensions from the tissues were prepared as described previously in ². The following antibodies were used: anti-CD19 (1D3, #45-0193-80, eBioscience), anti-B220 (RA3-6B2, #RM2630, Life Technologies), anti-CD45 (104, #109825, Biolegend), anti-CD1d (1B1, #123507, Biolegend), anti-CD140 (APA5, #135905, Biolegend), anti-CD21 (7E9, #123419, Biolegend), anti-CD5 (53-7.3, #100607, Biolegend), anti-AA4.1 (#17-5892, eBioscience), anti-CD138 (281-2, #142505, Biolegend), anti-CD206 (C068C2, Biolegend), anti-CD86 (GL-1, Biolegend), anti-F4-80 (BM8, Biolegend), anti-CD11b (M1-70, Biolegend). Dead cells were excluded by staining with Propidium Iodide (Sigma-Aldrich) or Aqua Live/Dead stain. Flow cytometry was performed on FACScalibur and LSRII II (BD Biosciences) instruments at NYU School of Medicine Flow Cytometry Core Facility and data was analyzed using FlowJo software.