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

PLoS One. 2013 Oct 30;8(10):e78408. doi: 10.1371/journal.pone.0078408. eCollection 2013.

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-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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antigens, CD19 / immunology
  • Antigens, Ly / immunology
  • B-Lymphocytes / immunology*
  • CD11c Antigen / immunology
  • Cell Differentiation / immunology
  • Cell Lineage / immunology
  • Dendritic Cells / immunology*
  • GPI-Linked Proteins / immunology
  • Leukocyte Common Antigens / immunology
  • Membrane Glycoproteins / immunology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Precursor Cells, B-Lymphoid / immunology*
  • Receptors, Complement / immunology
  • Receptors, Interleukin-7 / immunology

Substances

  • Antigens, CD19
  • Antigens, Ly
  • CD11c Antigen
  • GPI-Linked Proteins
  • Ly6d protein, mouse
  • Membrane Glycoproteins
  • Receptors, Complement
  • Receptors, Interleukin-7
  • complement 1q receptor
  • interleukin-7 receptor, alpha chain
  • Leukocyte Common Antigens