Anti ter119 biotin

T cells were magnetically enriched by negative selection from spleens of recipient mice following staining with anti-CD19-biotin (Fitch Monoclonal Facility), anti-Ter119-biotin (eBioscience), and anti-CD11b-biotin (eBioscience) and incubation with streptavidin magnetic beads (Pierce, Thermo Fisher Scientific). APCs were magnetically enriched by negative selection of splenocytes from CD45.1⁺ B6 mice following staining with anti-Thy1.2-biotin (eBioscience) and incubation with streptavidin magnetic beads (Pierce).

Spleen and lymph node cells were purified by forcing tissue through 40 μm mesh into complete RPMI media (Gibco) containing 6% serum. PE-specific B cells were enriched as previously described (Pape et al., 2011 (link); Taylor et al., 2012b (link)). To enrich plasma cells, cell suspensions were made from bone marrow in PBS containing 1 mM EDTA and 2% serum. Anti-CD138-APC (BD Biosciences) and anti-APC microbeads (Militenyi Biotec) were used for enrichment with magnetized LS columns. In figure 3, APC-specific cells were enriched by negative selection using anti-CD4-biotin, anti-CD8-biotin, anti-Gr1-biotin, and anti-Ter119-biotin (eBioscience), streptavidin microbeads (Militenyi Biotec), and magnetized LS columns. Following column-based enrichment, samples were processed for flow cytometry. Cell counts from column-bound and flow-through samples were assessed using AccuCheck counting beads (Invitrogen) and cell numbers from both fractions were evaluated.