Murine Plasmacytoid Pre-Dendritic Cells Generated from Flt3 Ligand-Supplemented Bone Marrow Cultures Are Immature APCs

AbstractHeme is an erythrocyte-derived toxin that drives disease progression in hemolytic anemias, such as sickle cell disease. During hemolysis, specialized bone marrow-derived macrophages with a high heme-metabolism capacity orchestrate disease adaptation by removing damaged erythrocytes and heme-protein complexes from the blood and supporting iron recycling for erythropoiesis. Since chronic heme-stress is noxious for macrophages, erythrophagocytes in the spleen are continuously replenished from bone marrow-derived progenitors. Here, we hypothesized that adaptation to heme stress progressively shifts differentiation trajectories of bone marrow progenitors to expand the capacity of heme-handling monocyte-derived macrophages at the expense of the homeostatic generation of dendritic cells, which emerge from shared myeloid precursors. This heme-induced redirection of differentiation trajectories may contribute to hemolysis-induced secondary immunodeficiency. We performed single-cell RNA-sequencing with directional RNA velocity analysis of GM-CSF-supplemented mouse bone marrow cultures to assess myeloid differentiation under heme stress. We found that heme-activated NRF2 signaling shifted the differentiation of bone marrow cells towards antioxidant, iron-recycling macrophages, suppressing the generation of dendritic cells in heme-exposed bone marrow cultures. Heme eliminated the capacity of GM-CSF-supplemented bone marrow cultures to activate antigen-specific CD4 T cells. The generation of functionally competent dendritic cells was restored by NRF2 loss. The heme-induced phenotype of macrophage expansion with concurrent dendritic cell depletion was reproduced in hemolytic mice with sickle cell disease and spherocytosis and associated with reduced dendritic cell functions in the spleen. Our data provide a novel mechanistic underpinning of hemolytic stress as a driver of hyposplenism-related secondary immunodeficiency.

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Differentiation of dendritic cells in cultures of rat bone marrow cells.

Journal of Experimental Medicine ◽

10.1084/jem.163.4.872 ◽

1986 ◽

Vol 163 (4) ◽

pp. 872-883 ◽

Cited By ~ 45

Author(s):

W E Bowers ◽

M R Berkowitz

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

Bone Marrow Cells ◽

Sensitive Assay ◽

Ia Antigens ◽

Bone Marrow Cultures ◽

Nonadherent Cells ◽

Marrow Cultures ◽

Rat Bone ◽

Marrow Cells

Although dendritic cells (DC) originate from bone marrow, they were not observed in fresh preparations of bone marrow cells (BMC). Likewise, accessory activity was barely measurable in a sensitive assay for this potent function of DC. However, both DC and accessory activity developed when BMC were cultured for 5 d. Based on fractionation before culture, nearly all of the accessory activity could be attributed to only 5% of the total BMC recovered in a low-density (LD) fraction. The LD-DC precursors differed from mature DC in a number of important respects. Removal of Ia+ cells from the LD fraction by panning did not decrease the production of DC when the nonadherent cells were cultured. Thus, the cell from which the DC is derived does not express or minimally expresses Ia antigens, in contrast to the strongly Ia+ DC that is produced in bone marrow cultures. Irradiation of LD cells before culture prevented the development of DC. When irradiation was delayed by daily intervals, progressive increases in the number of DC resulted, up to the fifth day. These findings, together with preliminary autoradiographic data, indicate that cell division has occurred, in contrast to the DC, which does not divide. We conclude that bone marrow-derived DC arise in culture from the division of LD, Ia- precursors.

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Generation or large numbers of immature and mature dendritic cells from rat bone marrow cultures

European Journal of Immunology ◽

10.1002/(sici)1521-4141(199803)28:03<811::aid-immu811>3.0.co;2-s ◽

1998 ◽

Vol 28 (3) ◽

pp. 811-817 ◽

Cited By ~ 83

Author(s):

Mia Talmor ◽

Asra Mirza ◽

Shannon Turley ◽

Ira Mellman ◽

Lloyd A. Hoffman ◽

...

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

Large Numbers ◽

Bone Marrow Cultures ◽

Marrow Cultures ◽

Rat Bone

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Strain-Specific Variations in the Development of Dendritic Cells in Murine Bone-Marrow Cultures

Scandinavian Journal of Immunology ◽

10.1046/j.1365-3083.2000.00732.x ◽

2000 ◽

Vol 51 (6) ◽

pp. 586-594 ◽

Cited By ~ 9

Author(s):

Petersen ◽

Toldbod ◽

Holtz ◽

Hokland ◽

Bolund ◽

...

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

Murine Bone Marrow ◽

Bone Marrow Cultures ◽

Marrow Cultures

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GM-CSF Mouse Bone Marrow Cultures Comprise a Heterogeneous Population of CD11c+MHCII+ Macrophages and Dendritic Cells

Immunity ◽

10.1016/j.immuni.2015.05.018 ◽

2015 ◽

Vol 42 (6) ◽

pp. 1197-1211 ◽

Cited By ~ 370

Author(s):

Julie Helft ◽

Jan Böttcher ◽

Probir Chakravarty ◽

Santiago Zelenay ◽

Jatta Huotari ◽

...

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

Heterogeneous Population ◽

Mouse Bone Marrow ◽

Gm Csf ◽

Bone Marrow Cultures ◽

Marrow Cultures

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Faculty Opinions recommendation of GM-CSF Mouse Bone Marrow Cultures Comprise a Heterogeneous Population of CD11c(+)MHCII(+) Macrophages and Dendritic Cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.725566020.793527829 ◽

2017 ◽

Author(s):

Kenneth Murphy

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

Heterogeneous Population ◽

Mouse Bone Marrow ◽

Gm Csf ◽

Bone Marrow Cultures ◽

Marrow Cultures

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The TH1 Lymphokine Interferon-γ is a Potent Upregulator of Dendritic Cells with Phagocytic Capacity in GM-CSF Supplemented Bone Marrow Cultures

Advances in Experimental Medicine and Biology - Dendritic Cells in Fundamental and Clinical Immunology ◽

10.1007/978-1-4757-9966-8_37 ◽

1997 ◽

pp. 221-225 ◽

Cited By ~ 1

Author(s):

Christoph Scheicher ◽

Heike Corban ◽

Vera Hof ◽

Michael Robbers ◽

Konrad Reske

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

Interferon Γ ◽

Phagocytic Capacity ◽

Gm Csf ◽

Bone Marrow Cultures ◽

Marrow Cultures

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Heme-stress activated NRF2 signaling skews fate trajectories of bone marrow cells from dendritic cells towards red pulp-like macrophages

10.1101/2021.07.29.454342 ◽

2021 ◽

Author(s):

Florence Vallelian ◽

Raphael M Buzzi ◽

Marc Pfefferle ◽

Ayla Yalamanoglu ◽

Andreas Wassmer ◽

...

Keyword(s):

Bone Marrow ◽

Dendritic Cells ◽

Dendritic Cell ◽

Sickle Cell Disease ◽

Bone Marrow Cells ◽

Cell Disease ◽

Secondary Immunodeficiency ◽

Gm Csf ◽

Bone Marrow Cultures ◽

Marrow Cultures

Heme is an erythrocyte-derived toxin that drives disease progression in hemolytic anemias, such as sickle cell disease. During hemolysis, specialized bone marrow-derived macrophages with a high heme-metabolism capacity orchestrate disease adaptation by removing damaged erythrocytes and heme-protein complexes from the blood and supporting iron recycling for erythropoiesis. Since chronic heme-stress is noxious for macrophages, erythrophagocytes in the spleen are continuously replenished from bone marrow-derived progenitors. Here, we hypothesized that adaptation to heme stress progressively shifts differentiation trajectories of BM progenitors to expand the capacity of heme-handling monocyte-derived macrophages at the expense of the homeostatic generation of dendritic cells, which emerge from shared myeloid precursors. This heme-induced redirection of differentiation trajectories may contribute to hemolysis-induced secondary immunodeficiency. We performed single-cell RNA sequencing with directional RNA velocity analysis of GM-CSF-supplemented mouse bone marrow cultures to assess myeloid differentiation under heme stress. We found that heme-activated NRF2 signaling shifted the differentiation of bone marrow cells towards antioxidant, iron-recycling macrophages, suppressing the generation of dendritic cells in heme-exposed bone marrow cultures. Heme eliminated the capacity of GM-CSF-supplemented bone marrow cultures to activate antigen-specific CD4 T cells. The generation of functionally competent dendritic cells was restored by NRF2 loss. The heme-induced phenotype of macrophage expansion with concurrent dendritic cell depletion was reproduced in hemolytic mice with sickle cell disease and spherocytosis and associated with reduced dendritic cell functions in the spleen. Our data provide a novel mechanistic underpinning of hemolytic stress as a driver of hyposplenism-related secondary immunodeficiency.

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