TY - JOUR

T1 - Ascorbic acid supports ex vivo generation of plasmacytoid dendritic cells from circulating hematopoietic stem cells

AU - Laustsen, Anders

AU - van der Sluis, Renée Marije

AU - Gris Oliver, Albert

AU - Sanchez Hernandez, Sabina

AU - Cemalovic, Ena

AU - Tang, Hai Qing

AU - Pedersen, Lars Henning

AU - Uldbjerg, Niels

AU - Jakobsen, Martin Roelsgaard

AU - Bak, Rasmus

PY - 2021/9

Y1 - 2021/9

N2 - Plasmacytoid dendritic cells (pDCs) constitute a rare type of immune cell with multifaceted functions, but their potential use as a cell-based immunotherapy is challenged by the scarce cell numbers that can be extracted from blood. Here, we systematically investigate culture parameters for generating pDCs from hematopoietic stem and progenitor cells (HSPCs). Using optimized conditions combined with implementation of HSPC pre-expansion, we generate an average of 465 million HSPC-derived pDCs (HSPC-pDCs) starting from 100,000 cord blood-derived HSPCs. Furthermore, we demonstrate that such protocol allows HSPC-pDC generation from whole blood HSPCs, and these cells display a pDC phenotype and function. Using GMP compliant medium, we observe a remarkable loss of TLR7/9 responses, which is rescued by ascorbic acid supplementation. Ascorbic acid induces transcriptional signatures associated with pDC-specific innate immune pathways suggesting an undescribed role of ascorbic acid for pDC functionality. This constitutes the first protocol for generating pDCs from whole blood, and lay the foundation for investigating HSPC-pDCs for cell-based immunotherapy

AB - Plasmacytoid dendritic cells (pDCs) constitute a rare type of immune cell with multifaceted functions, but their potential use as a cell-based immunotherapy is challenged by the scarce cell numbers that can be extracted from blood. Here, we systematically investigate culture parameters for generating pDCs from hematopoietic stem and progenitor cells (HSPCs). Using optimized conditions combined with implementation of HSPC pre-expansion, we generate an average of 465 million HSPC-derived pDCs (HSPC-pDCs) starting from 100,000 cord blood-derived HSPCs. Furthermore, we demonstrate that such protocol allows HSPC-pDC generation from whole blood HSPCs, and these cells display a pDC phenotype and function. Using GMP compliant medium, we observe a remarkable loss of TLR7/9 responses, which is rescued by ascorbic acid supplementation. Ascorbic acid induces transcriptional signatures associated with pDC-specific innate immune pathways suggesting an undescribed role of ascorbic acid for pDC functionality. This constitutes the first protocol for generating pDCs from whole blood, and lay the foundation for investigating HSPC-pDCs for cell-based immunotherapy

KW - ACTIVATION

KW - BONE-MARROW

KW - CANCER

KW - Human

KW - I INTERFERON-PRODUCTION

KW - PERIPHERAL-BLOOD

KW - PROGENITOR CELLS

KW - SPI-B

KW - T-CELLS

KW - TLR7

KW - TLR9

KW - TRANSCRIPTION FACTOR E2-2

KW - UMBILICAL-CORD BLOOD

KW - ascorbic acid

KW - cHSPC

KW - dendritic cells

KW - differentiation

KW - ex

KW - hematopoietic stem cells

KW - immunotherapy

KW - pDC

KW - plasmacytoid

KW - vitamin c

KW - vivo

UR - http://www.scopus.com/inward/record.url?scp=85116363217&partnerID=8YFLogxK

U2 - 10.7554/eLife.65528

DO - 10.7554/eLife.65528

M3 - Journal article

C2 - 34473049

AN - SCOPUS:85116363217

VL - 10

JO - eLife

JF - eLife

SN - 2050-084X

M1 - e65528

ER -