Open AccessSeparation of Plasmacytoid Dendritic Cells from B-Cell-Biased Lymphoid Progenitor (BLP) and Pre-Pro B Cells Using PDCA-1
Author(s) -
Kay L. Medina,
Sarah Tangen,
Lauren Seaburg,
Puspa Thapa,
Kimberly A. Gwin,
Virginia Smith Shapiro
Publication year - 2013
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0078408
Subject(s) - progenitor cell , cd19 , b cell , pdca , biology , microbiology and biotechnology , cd11c , lymphopoiesis , dendritic cell , stem cell , immunology , flow cytometry , phenotype , antibody , antigen , genetics , management system , management , gene , economics , quality management
B-cell-biased lymphoid progenitors (BLPs) and Pre-pro B cells lie at a critical juncture between B cell specification and commitment. However, both of these populations are heterogenous, which hampers investigation into the molecular changes that occur as lymphoid progenitors commit to the B cell lineage. Here, we demonstrate that there are PDCA-1 + Siglec H + plasmacytoid dendritic cells (pDCs) that co-purify with BLPs and Pre-pro B cells, which express little or no CD11c or Ly6C. Removal of PDCA-1 + pDCs separates B cell progenitors that express high levels of a Rag1-GFP reporter from Rag1-GFP low/neg pDCs within the BLP and Pre-pro B populations. Analysis of Flt3-ligand knockout and IL-7Rα knockout mice revealed that there is a block in B cell development at the all-lymphoid progenitor (ALP) stage, as the majority of cells within the BLP or Pre-pro B gates were PDCA-1 + pDCs. Thus, removal of PDCA-1 + pDCs is critical for analysis of BLP and Pre-pro B cell populations. Analysis of B cell potential within the B220 + CD19 − fraction demonstrated that AA4.1 + Ly6D + PDCA-1 − Pre-pro B cells gave rise to CD19 + B cells at high frequency, while PDCA-1 + pDCs in this fraction did not. Interestingly, the presence of PDCA-1 + pDCs within CLPs may help to explain the conflicting results regarding the origin of these cells.