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

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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Alive but Confused: Heterogeneity of CD11c + MHC Class II + Cells in GM-CSF Mouse Bone Marrow Cultures

Immunity ◽

10.1016/j.immuni.2015.12.014 ◽

2016 ◽

Vol 44 (1) ◽

pp. 3-4 ◽

Cited By ~ 13

Author(s):

Julie Helft ◽

Jan P. Böttcher ◽

Probir Chakravarty ◽

Santiago Zelenay ◽

Jatta Huotari ◽

...

Keyword(s):

Bone Marrow ◽

Mhc Class Ii ◽

Class Ii ◽

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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Early pre-B-cell transformation induced by the v-fms oncogene in long-term mouse bone marrow cultures.

Molecular and Cellular Biology ◽

10.1128/mcb.9.9.3973 ◽

1989 ◽

Vol 9 (9) ◽

pp. 3973-3981 ◽

Cited By ~ 15

Author(s):

G V Borzillo ◽

C J Sherr

Keyword(s):

Bone Marrow ◽

B Cells ◽

Chain Gene ◽

Mouse Bone Marrow ◽

Feeder Layers ◽

Bone Marrow Cultures ◽

B Lymphoid ◽

Marrow Cultures

Murine long-term bone marrow cultures that support B-lymphoid-cell development were infected with a helper-free retrovirus containing the v-fms oncogene. Infection of B-lymphoid cultures resulted in the rapid clonal outgrowth of early pre-B cells, which grew to high cell densities on stromal cell feeder layers, expressed v-fms-coded glycoproteins, and underwent immunoglobulin heavy-chain gene rearrangements. Late-passage cultures gave rise to factor-independent variants that proliferated in the absence of feeder layers, developed resistance to hydrocortisone, and became tumorigenic in syngeneic mice. The v-fms oncogene therefore recapitulates known effects of the v-abl and bcr-abl oncogenes on B-lineage cells. The ability of v-fms to induce transformation of early pre-B cells in vitro underscores the capacity of oncogenic mutants of the colony-stimulating factor-1 receptor to function outside the mononuclear phagocyte lineage.

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Early pre-B-cell transformation induced by the v-fms oncogene in long-term mouse bone marrow cultures

Molecular and Cellular Biology ◽

10.1128/mcb.9.9.3973-3981.1989 ◽

1989 ◽

Vol 9 (9) ◽

pp. 3973-3981

Author(s):

G V Borzillo ◽

C J Sherr

Keyword(s):

Bone Marrow ◽

B Cells ◽

Chain Gene ◽

Mouse Bone Marrow ◽

Feeder Layers ◽

Bone Marrow Cultures ◽

B Lymphoid ◽

Marrow Cultures

Murine long-term bone marrow cultures that support B-lymphoid-cell development were infected with a helper-free retrovirus containing the v-fms oncogene. Infection of B-lymphoid cultures resulted in the rapid clonal outgrowth of early pre-B cells, which grew to high cell densities on stromal cell feeder layers, expressed v-fms-coded glycoproteins, and underwent immunoglobulin heavy-chain gene rearrangements. Late-passage cultures gave rise to factor-independent variants that proliferated in the absence of feeder layers, developed resistance to hydrocortisone, and became tumorigenic in syngeneic mice. The v-fms oncogene therefore recapitulates known effects of the v-abl and bcr-abl oncogenes on B-lineage cells. The ability of v-fms to induce transformation of early pre-B cells in vitro underscores the capacity of oncogenic mutants of the colony-stimulating factor-1 receptor to function outside the mononuclear phagocyte lineage.

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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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Diethyldithiocarbamate induction of cytokine release in human long-term bone marrow cultures

Blood ◽

10.1182/blood.v80.5.1172.1172 ◽

1992 ◽

Vol 80 (5) ◽

pp. 1172-1177 ◽

Cited By ~ 14

Author(s):

CJ East ◽

CN Abboud ◽

RF Borch

Keyword(s):

Bone Marrow ◽

Conditioned Medium ◽

Chemotherapeutic Agents ◽

Tnf Alpha ◽

Complete Medium ◽

Maximum Increase ◽

Gm Csf ◽

Bone Marrow Cultures ◽

Marrow Cultures

Abstract Diethyldithiocarbamate (DDTC) is a biochemical modulating agent that protects murine bone marrow progenitor cells from the cytotoxicity of a variety of cancer chemotherapeutic agents. However, the mechanism of this protection is not well understood. Long-term human bone marrow cultures (LTBMC) were established and at day 17 treated with 30 mumol/L DDTC for 1 hour, after which DDTC was removed and replaced with complete medium. Conditioned medium was then collected 6, 12, 24, and 48 hours later and analyzed for the presence of cytokines. A time- dependent increase in granulocyte-macrophage colony-stimulating factor (GM-CSF) (12-fold), granulocyte-CSF (G-CSF) (66-fold), interleukin (IL)- 6, (three-fold), IL-1 beta (161-fold), and tumor necrosis factor (TNF)- alpha (25-fold) was observed. The maximum increase for the factors other than TNF-alpha was at 24 to 48 hours posttreatment. However, TNF- alpha peaked as early as 6 hours post-DDTC. When conditioned medium from these cultures was tested in a granulocyte-macrophage progenitor cell (GM-CFC) assay, an increase in colony formation was observed that correlated with the increased levels of cytokines in the medium. The specificity of this effect was confirmed by the fact that the closely related congener bis(hydroxyethyl)dithiocarbamate was devoid of colony- stimulating activity. The addition of antibodies for TNF-alpha and/or IL-1 alpha following DDTC treatment did not inhibit the release of GM- CSF, G-CSF, or IL-6 from the LTBMC. These results suggest that DDTC accelerates bone marrow recovery following myelotoxic drug treatment via increased production of cytokines that are known to be essential for hematopoiesis.

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