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Related Experiment Video

Updated: Dec 13, 2025

Study of Dendritic Cell Development by Short Hairpin RNA-Mediated Gene Knockdown in a Hematopoietic Stem and Progenitor Cell Line In vitro
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Differential IRF8 Transcription Factor Requirement Defines Two Pathways of Dendritic Cell Development in Humans.

Urszula Cytlak1, Anastasia Resteu1, Sarah Pagan1

  • 1Translational and Clinical Research Institute, Newcastle University, Newcastle upon Tyne NE2 4HH, UK.

Immunity
|August 1, 2020
PubMed
Summary

IRF8 transcription factor controls dendritic cell (DC) development. This study reveals two distinct pathways for DC subset heterogeneity, impacting DC2 and DC3 cell lineages in humans with IRF8 deficiency.

Keywords:
CyTOFIRF8dendritic cellhematopoiesisimmunityprimary immunodeficiencysingle-cell RNA sequencingtranscription factor

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Area of Science:

  • Immunology
  • Developmental Biology
  • Hematopoiesis

Background:

  • Dendritic cell (DC) formation is regulated by hematopoietic transcription factors, notably IRF8.
  • IRF8 deficiency differentially impacts DC subsets, including plasmacytoid DCs (pDCs) and classical DCs (cDC1, cDC2).
  • Human cDC2-related subsets (AXL+SIGLEC6+ pre-DC, DC2, DC3) exhibit heterogeneity with unknown origins.

Purpose of the Study:

  • To elucidate the developmental origins of cDC2 heterogeneity.
  • To investigate the role of IRF8 in distinct DC developmental pathways.
  • To understand the impact of IRF8 deficiency on human DC subsets.

Main Methods:

  • High-dimensional analysis of human immune cells.
  • In vitro differentiation of hematopoietic progenitors.
  • Utilizing an allelic series of human IRF8 deficiency.

Main Results:

  • Identified two distinct developmental pathways for cDC2 heterogeneity: an IRF8hi lymphoid-primed pathway and an IRF8lo common myeloid pathway.
  • The IRF8hi pathway supports pDC, cDC1, and DC2 trajectories, while the IRF8lo pathway generates DC3s and monocytes.
  • AXL+SIGLEC6+ pre-DCs exclusively map to the DC2 pathway.
  • DC3s expand and replace DC2s in partial IRF8 deficiency due to lower IRF8 requirement.

Conclusions:

  • Human cDC2 heterogeneity arises from two distinct developmental routes influenced by IRF8 levels.
  • IRF8 expression level is critical in directing hematopoietic progenitors towards specific DC lineages.
  • Understanding these pathways provides insight into DC development and the consequences of IRF8 deficiency.