Abstract
Plasmacytoid dendritic cells (pDC) are essential for immune competence. Here we show that pDC precursor differentiated from human CD34+ hematopoietic stem and progenitor cells (HSPC) has low surface expression of pDC markers, and has limited induction of type I interferon (IFN) and IL-6 upon TLR7 and TLR9 agonists treatment; by contrast, cGAS or RIG-I agonists-mediated activation is not altered. Importantly, after priming with type I and II IFN, these precursor pDCs attain a phenotype and functional activity similar to that of peripheral blood-derived pDCs. Data from CRISPR/Cas9-mediated genome editing of HSPCs further show that HSPC-pDCs with genetic modifications can be obtained, and that expression of the IFN-α receptor is essential for the optimal function, but dispensable for the differentiation, of HSPC-pDC percursor. Our results thus demonstrate the biological effects of IFNs for regulating pDC function, and provide the means of generating of gene-modified human pDCs.
Highlights
Plasmacytoid dendritic cells are essential for immune competence
Plasmacytoid dendritic cells are key effectors in cellular immunity with the ability to initiate immune responses and to induce tolerance to exogenous and endogenous antigens1. pDCs are distinct from conventional DCs as they express high levels of interferon regulatory factor 7 (IRF7) and they primarily sense pathogens through Toll-like receptor (TLR) 7 and 91,2
This is an important benefit of this culture system, since inhibition of gene expression in by RNAi-based methods in pDCs can be problematic due to non-specific immune activation through, e.g. RIG-I or TLR3/7/815,40–42
Summary
Plasmacytoid dendritic cells (pDC) are essential for immune competence. Here we show that pDC precursor differentiated from human CD34+ hematopoietic stem and progenitor cells (HSPC) has low surface expression of pDC markers, and has limited induction of type I interferon (IFN) and IL-6 upon TLR7 and TLR9 agonists treatment; by contrast, cGAS or RIG-I agonists-mediated activation is not altered. After priming with type I and II IFN, these precursor pDCs attain a phenotype and functional activity similar to that of peripheral blood-derived pDCs. Data from CRISPR/Cas9-mediated genome editing of HSPCs further show that HSPC-pDCs with genetic modifications can be obtained, and that expression of the IFN-α receptor is essential for the optimal function, but dispensable for the differentiation, of HSPC-pDC percursor. Our results demonstrate the biological effects of IFNs for regulating pDC function, and provide the means of generating of gene-modified human pDCs. Plasmacytoid dendritic cells (pDCs) are key effectors in cellular immunity with the ability to initiate immune responses and to induce tolerance to exogenous and endogenous antigens. IFNs appear to regulate key aspects of murine pDC biology, including development, apoptosis, migration, homeostasis, and function Their role in dictating human pDC development and functions remains incompletely understood. We demonstrate that genetically modified pDCs can be derived from CRISPR/Cas[9] gene-edited CD34+ HSPCs, and that knockout of the type I IFN receptor abrogates TLR7 and TLR9-mediated innate sensing
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