Separation of plasmacytoid dendritic cells from B-cell-biased lymphoid progenitor (BLP) and pre-pro B cells using PDCA-1

Kay L. Medina, Sarah N. Tangen, Lauren M. Seaburg, Puspa Thapa, Kimberly A. Gwin, Virginia Smith Shapiro

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

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-GFPow/neg pDCs within the BLP and Pre-pro B populations. Analysis of Flt3-ligand knockout and IL-7Ra 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.

Original languageEnglish (US)
Article numbere78408
JournalPloS one
Volume8
Issue number10
DOIs
StatePublished - Oct 30 2013

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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