Large-Scale Human Dendritic Cell Differentiation Revealing Notch-Dependent Lineage Bifurcation and Heterogeneity

Cell Rep. 2018 Aug 14;24(7):1902-1915.e6. doi: 10.1016/j.celrep.2018.07.033.

Abstract

The ability to generate large numbers of distinct types of human dendritic cells (DCs) in vitro is critical for accelerating our understanding of DC biology and harnessing them clinically. We developed a DC differentiation method from human CD34+ precursors leading to high yields of plasmacytoid DCs (pDCs) and both types of conventional DCs (cDC1s and cDC2s). The identity of the cells generated in vitro and their strong homology to their blood counterparts were demonstrated by phenotypic, functional, and single-cell RNA-sequencing analyses. This culture system revealed a critical role of Notch signaling and GM-CSF for promoting cDC1 generation. Moreover, we discovered a pre-terminal differentiation state for each DC type, characterized by high expression of cell-cycle genes and lack of XCR1 in the case of cDC1. Our culture system will greatly facilitate the simultaneous and comprehensive study of primary, otherwise rare human DC types, including their mutual interactions.

Keywords: CLEC10A; CLEC9A; NOTCH; XCR1; adjuvant; dendritic cell differentiation; dendritic cell types; hematopoiesis; immunotherapy; plasmacytoid dendritic cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD34 / genetics
  • Antigens, CD34 / immunology
  • Calcium-Binding Proteins
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / immunology
  • Cell Differentiation / drug effects
  • Cell Lineage / immunology*
  • Dendritic Cells / cytology
  • Dendritic Cells / drug effects
  • Dendritic Cells / immunology*
  • Gene Expression
  • Gene Expression Profiling
  • Granulocyte-Macrophage Colony-Stimulating Factor / genetics*
  • Granulocyte-Macrophage Colony-Stimulating Factor / immunology
  • Humans
  • Imidazoles / pharmacology
  • Immunophenotyping
  • Intercellular Signaling Peptides and Proteins / genetics*
  • Intercellular Signaling Peptides and Proteins / immunology
  • Lipopolysaccharides / pharmacology
  • Membrane Proteins / genetics*
  • Membrane Proteins / immunology
  • Poly I-C / pharmacology
  • Primary Cell Culture
  • Receptor, Notch1 / genetics*
  • Receptor, Notch1 / immunology
  • Receptors, G-Protein-Coupled / deficiency
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / immunology
  • Signal Transduction
  • Single-Cell Analysis

Substances

  • Antigens, CD34
  • Calcium-Binding Proteins
  • Cell Cycle Proteins
  • DLK1 protein, human
  • Imidazoles
  • Intercellular Signaling Peptides and Proteins
  • Lipopolysaccharides
  • Membrane Proteins
  • NOTCH1 protein, human
  • Receptor, Notch1
  • Receptors, G-Protein-Coupled
  • XCR1 protein, human
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Poly I-C
  • resiquimod