Monocyte-derived macrophages matured under prolonged hypoxia transcriptionally up-regulate HIF-1α mRNA

Karl J Staples, Fattah Sotoodehnejadnematalahi, Helen Pearson, Marion Frankenberger, Lorenza Francescut, Loems Ziegler-Heitbrock, Bernard Burke

Research output: Contribution to journalArticle

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Abstract

This study tested the hypothesis that prolonged severe hypoxia during monocyte to macrophage differentiation results in macrophages with a pattern of gene expression and phenotype distinct from those maturing in normal oxygen levels. Macrophages accumulate in hypoxic and anoxic areas within pathological sites such as tumours, wounds, and arthritic joints, and have been proposed as vehicles for gene therapy delivery to such tissues. Several non-pathological tissues are also hypoxic. We therefore argue that differentiation from monocyte to macrophage in hypoxic conditions is a common occurrence. However, the effect of long term severe hypoxia on monocyte to macrophage differentiation has not been studied. Here, using primary human peripheral blood monocytes, we show that maturation for 5 days in 0.2% oxygen results in decreased phagocytosis, and decreased CD40 and CD206 expression. Chronic hypoxia induced much higher mRNA levels of the pro-angiogenic cytokine, VEGF, in adherence-purified macrophages (27-fold), CD14-magnetic bead purified monocytes (90-fold), and PBMC (104-fold) compared to acute (24h) hypoxia (11, 17 and 9-fold, respectively). This suggests that macrophages may play an even greater role in angiogenesis than previously appreciated. Furthermore, chronic hypoxia resulted in up-regulation of HIF-1α mRNA, in all monocyte-derived macrophage types studied. Actinomycin D experiments indicate that the increases in HIF-1α mRNA were not due to increased mRNA stability. To our knowledge this is the first study demonstrating up-regulation of HIF-1α mRNA by hypoxia in macrophages. Taken together, the data support the hypothesis that hypoxia affects monocyte to macrophage maturation, resulting in a distinct gene expression pattern and phenotype.

Original languageEnglish
Pages (from-to)832-839
Number of pages8
JournalImmunobiology
Volume216
Issue number7
DOIs
Publication statusPublished - Jul 2011

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Up-Regulation
Macrophages
Messenger RNA
Monocytes
Hypoxia
Oxygen
Phenotype
Gene Expression
RNA Stability
Dactinomycin
Phagocytosis
Genetic Therapy
Vascular Endothelial Growth Factor A
Arthritis
Joints
Cytokines
Wounds and Injuries
Neoplasms

Keywords

  • Antigens, CD/genetics
  • Cell Differentiation/drug effects
  • Cells, Cultured
  • Humans
  • Hypoxia/genetics
  • Hypoxia-Inducible Factor 1, alpha Subunit/genetics
  • Macrophages/drug effects
  • Oxygen/pharmacology
  • Phagocytosis/drug effects
  • RNA, Messenger/analysis
  • Up-Regulation
  • Vascular Endothelial Growth Factor A/genetics

Cite this

Staples, K. J., Sotoodehnejadnematalahi, F., Pearson, H., Frankenberger, M., Francescut, L., Ziegler-Heitbrock, L., & Burke, B. (2011). Monocyte-derived macrophages matured under prolonged hypoxia transcriptionally up-regulate HIF-1α mRNA. Immunobiology, 216(7), 832-839. https://doi.org/10.1016/j.imbio.2010.12.005

Monocyte-derived macrophages matured under prolonged hypoxia transcriptionally up-regulate HIF-1α mRNA. / Staples, Karl J; Sotoodehnejadnematalahi, Fattah; Pearson, Helen; Frankenberger, Marion; Francescut, Lorenza; Ziegler-Heitbrock, Loems; Burke, Bernard.

In: Immunobiology, Vol. 216, No. 7, 07.2011, p. 832-839.

Research output: Contribution to journalArticle

Staples, KJ, Sotoodehnejadnematalahi, F, Pearson, H, Frankenberger, M, Francescut, L, Ziegler-Heitbrock, L & Burke, B 2011, 'Monocyte-derived macrophages matured under prolonged hypoxia transcriptionally up-regulate HIF-1α mRNA' Immunobiology, vol. 216, no. 7, pp. 832-839. https://doi.org/10.1016/j.imbio.2010.12.005
Staples KJ, Sotoodehnejadnematalahi F, Pearson H, Frankenberger M, Francescut L, Ziegler-Heitbrock L et al. Monocyte-derived macrophages matured under prolonged hypoxia transcriptionally up-regulate HIF-1α mRNA. Immunobiology. 2011 Jul;216(7):832-839. https://doi.org/10.1016/j.imbio.2010.12.005
Staples, Karl J ; Sotoodehnejadnematalahi, Fattah ; Pearson, Helen ; Frankenberger, Marion ; Francescut, Lorenza ; Ziegler-Heitbrock, Loems ; Burke, Bernard. / Monocyte-derived macrophages matured under prolonged hypoxia transcriptionally up-regulate HIF-1α mRNA. In: Immunobiology. 2011 ; Vol. 216, No. 7. pp. 832-839.
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AU - Pearson, Helen

AU - Frankenberger, Marion

AU - Francescut, Lorenza

AU - Ziegler-Heitbrock, Loems

AU - Burke, Bernard

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N2 - This study tested the hypothesis that prolonged severe hypoxia during monocyte to macrophage differentiation results in macrophages with a pattern of gene expression and phenotype distinct from those maturing in normal oxygen levels. Macrophages accumulate in hypoxic and anoxic areas within pathological sites such as tumours, wounds, and arthritic joints, and have been proposed as vehicles for gene therapy delivery to such tissues. Several non-pathological tissues are also hypoxic. We therefore argue that differentiation from monocyte to macrophage in hypoxic conditions is a common occurrence. However, the effect of long term severe hypoxia on monocyte to macrophage differentiation has not been studied. Here, using primary human peripheral blood monocytes, we show that maturation for 5 days in 0.2% oxygen results in decreased phagocytosis, and decreased CD40 and CD206 expression. Chronic hypoxia induced much higher mRNA levels of the pro-angiogenic cytokine, VEGF, in adherence-purified macrophages (27-fold), CD14-magnetic bead purified monocytes (90-fold), and PBMC (104-fold) compared to acute (24h) hypoxia (11, 17 and 9-fold, respectively). This suggests that macrophages may play an even greater role in angiogenesis than previously appreciated. Furthermore, chronic hypoxia resulted in up-regulation of HIF-1α mRNA, in all monocyte-derived macrophage types studied. Actinomycin D experiments indicate that the increases in HIF-1α mRNA were not due to increased mRNA stability. To our knowledge this is the first study demonstrating up-regulation of HIF-1α mRNA by hypoxia in macrophages. Taken together, the data support the hypothesis that hypoxia affects monocyte to macrophage maturation, resulting in a distinct gene expression pattern and phenotype.

AB - This study tested the hypothesis that prolonged severe hypoxia during monocyte to macrophage differentiation results in macrophages with a pattern of gene expression and phenotype distinct from those maturing in normal oxygen levels. Macrophages accumulate in hypoxic and anoxic areas within pathological sites such as tumours, wounds, and arthritic joints, and have been proposed as vehicles for gene therapy delivery to such tissues. Several non-pathological tissues are also hypoxic. We therefore argue that differentiation from monocyte to macrophage in hypoxic conditions is a common occurrence. However, the effect of long term severe hypoxia on monocyte to macrophage differentiation has not been studied. Here, using primary human peripheral blood monocytes, we show that maturation for 5 days in 0.2% oxygen results in decreased phagocytosis, and decreased CD40 and CD206 expression. Chronic hypoxia induced much higher mRNA levels of the pro-angiogenic cytokine, VEGF, in adherence-purified macrophages (27-fold), CD14-magnetic bead purified monocytes (90-fold), and PBMC (104-fold) compared to acute (24h) hypoxia (11, 17 and 9-fold, respectively). This suggests that macrophages may play an even greater role in angiogenesis than previously appreciated. Furthermore, chronic hypoxia resulted in up-regulation of HIF-1α mRNA, in all monocyte-derived macrophage types studied. Actinomycin D experiments indicate that the increases in HIF-1α mRNA were not due to increased mRNA stability. To our knowledge this is the first study demonstrating up-regulation of HIF-1α mRNA by hypoxia in macrophages. Taken together, the data support the hypothesis that hypoxia affects monocyte to macrophage maturation, resulting in a distinct gene expression pattern and phenotype.

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KW - Phagocytosis/drug effects

KW - RNA, Messenger/analysis

KW - Up-Regulation

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JO - Immunobiology

JF - Immunobiology

SN - 0171-2985

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