Increased TNF expression in CD43++ murine blood monocytes

Bernard Burke, Rasheedah Ahmad, Karl J Staples, Roger Snowden, Aras Kadioglu, Marion Frankenberger, David A Hume, Loems Ziegler-Heitbrock

Research output: Contribution to journalArticle

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Abstract

Monocyte heterogeneity has been studied extensively in man but only recently tools have been developed to study blood monocyte populations in the mouse. We have used the MacGreen mouse model, which expresses the green fluorescent protein under the control of the promoter of the murine M-CSF receptor (CSF1 receptor, c-fms). Since both monocytes and granulocytes show GFP expression in this model the latter cells were excluded by staining with the Ly6G granulocyte marker. GFP+ Ly6G- blood monocytes were found to account for an average of 246+/-121cells/microl in these mice. These monocytes can be subdivided into CD43+ GR-1+ cells and CD43++ GR-1(-) cells, with the latter cells accounting for 140+/-77cells/mul, i.e. about 60% of all blood monocytes. After intraperitoneal injection of lipopolysaccharide (LPS) both blood monocyte subpopulations were depleted. The same was true after intranasal infection with Streptococcus pneumoniae but here the CD43++ subpopulation was preferentially reduced to 4cells/mul. For the study of TNF expression cells were stimulated in vitro with LPS from Salmonella abortus equi in the presence of Brefeldin A followed by intracellular staining and multicolor flow cytometry. Over a dose range of 10-100ng LPS/ml, TNF protein production was significantly higher in the CD43++ monocyte subset. At 1000ng LPS/ml 90% of all CD43++ monocytes stained positive for TNF and in terms of fluorescence intensity TNF was 5-fold higher compared to the CD43+ monocytes. These data indicate that the murine CD43++ monocyte subset exhibits features of pro-inflammatory monocytes and is functionally homologeous to the human CD14+CD16+ monocytes.

Original languageEnglish
Pages (from-to)142-147
Number of pages6
JournalImmunology Letters
Volume118
Issue number2
DOIs
Publication statusPublished - 30 Jun 2008
Externally publishedYes

Fingerprint

Monocytes
Lipopolysaccharides
Granulocytes
Macrophage Colony-Stimulating Factor Receptors
Staining and Labeling
Brefeldin A
Pneumococcal Infections
Green Fluorescent Proteins
Intraperitoneal Injections
Salmonella
Flow Cytometry
Fluorescence

Keywords

  • Animals
  • Leukosialin/genetics
  • Lipopolysaccharides/pharmacology
  • Mice
  • Mice, Inbred C57BL
  • Monocytes/drug effects
  • Streptococcus pneumoniae/immunology
  • Tumor Necrosis Factor-alpha/genetics
  • Up-Regulation

Cite this

Burke, B., Ahmad, R., Staples, K. J., Snowden, R., Kadioglu, A., Frankenberger, M., ... Ziegler-Heitbrock, L. (2008). Increased TNF expression in CD43++ murine blood monocytes. Immunology Letters, 118(2), 142-147. https://doi.org/10.1016/j.imlet.2008.03.012

Increased TNF expression in CD43++ murine blood monocytes. / Burke, Bernard; Ahmad, Rasheedah; Staples, Karl J; Snowden, Roger; Kadioglu, Aras; Frankenberger, Marion; Hume, David A; Ziegler-Heitbrock, Loems.

In: Immunology Letters, Vol. 118, No. 2, 30.06.2008, p. 142-147.

Research output: Contribution to journalArticle

Burke, B, Ahmad, R, Staples, KJ, Snowden, R, Kadioglu, A, Frankenberger, M, Hume, DA & Ziegler-Heitbrock, L 2008, 'Increased TNF expression in CD43++ murine blood monocytes' Immunology Letters, vol. 118, no. 2, pp. 142-147. https://doi.org/10.1016/j.imlet.2008.03.012
Burke B, Ahmad R, Staples KJ, Snowden R, Kadioglu A, Frankenberger M et al. Increased TNF expression in CD43++ murine blood monocytes. Immunology Letters. 2008 Jun 30;118(2):142-147. https://doi.org/10.1016/j.imlet.2008.03.012
Burke, Bernard ; Ahmad, Rasheedah ; Staples, Karl J ; Snowden, Roger ; Kadioglu, Aras ; Frankenberger, Marion ; Hume, David A ; Ziegler-Heitbrock, Loems. / Increased TNF expression in CD43++ murine blood monocytes. In: Immunology Letters. 2008 ; Vol. 118, No. 2. pp. 142-147.
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abstract = "Monocyte heterogeneity has been studied extensively in man but only recently tools have been developed to study blood monocyte populations in the mouse. We have used the MacGreen mouse model, which expresses the green fluorescent protein under the control of the promoter of the murine M-CSF receptor (CSF1 receptor, c-fms). Since both monocytes and granulocytes show GFP expression in this model the latter cells were excluded by staining with the Ly6G granulocyte marker. GFP+ Ly6G- blood monocytes were found to account for an average of 246+/-121cells/microl in these mice. These monocytes can be subdivided into CD43+ GR-1+ cells and CD43++ GR-1(-) cells, with the latter cells accounting for 140+/-77cells/mul, i.e. about 60{\%} of all blood monocytes. After intraperitoneal injection of lipopolysaccharide (LPS) both blood monocyte subpopulations were depleted. The same was true after intranasal infection with Streptococcus pneumoniae but here the CD43++ subpopulation was preferentially reduced to 4cells/mul. For the study of TNF expression cells were stimulated in vitro with LPS from Salmonella abortus equi in the presence of Brefeldin A followed by intracellular staining and multicolor flow cytometry. Over a dose range of 10-100ng LPS/ml, TNF protein production was significantly higher in the CD43++ monocyte subset. At 1000ng LPS/ml 90{\%} of all CD43++ monocytes stained positive for TNF and in terms of fluorescence intensity TNF was 5-fold higher compared to the CD43+ monocytes. These data indicate that the murine CD43++ monocyte subset exhibits features of pro-inflammatory monocytes and is functionally homologeous to the human CD14+CD16+ monocytes.",
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AU - Burke, Bernard

AU - Ahmad, Rasheedah

AU - Staples, Karl J

AU - Snowden, Roger

AU - Kadioglu, Aras

AU - Frankenberger, Marion

AU - Hume, David A

AU - Ziegler-Heitbrock, Loems

PY - 2008/6/30

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N2 - Monocyte heterogeneity has been studied extensively in man but only recently tools have been developed to study blood monocyte populations in the mouse. We have used the MacGreen mouse model, which expresses the green fluorescent protein under the control of the promoter of the murine M-CSF receptor (CSF1 receptor, c-fms). Since both monocytes and granulocytes show GFP expression in this model the latter cells were excluded by staining with the Ly6G granulocyte marker. GFP+ Ly6G- blood monocytes were found to account for an average of 246+/-121cells/microl in these mice. These monocytes can be subdivided into CD43+ GR-1+ cells and CD43++ GR-1(-) cells, with the latter cells accounting for 140+/-77cells/mul, i.e. about 60% of all blood monocytes. After intraperitoneal injection of lipopolysaccharide (LPS) both blood monocyte subpopulations were depleted. The same was true after intranasal infection with Streptococcus pneumoniae but here the CD43++ subpopulation was preferentially reduced to 4cells/mul. For the study of TNF expression cells were stimulated in vitro with LPS from Salmonella abortus equi in the presence of Brefeldin A followed by intracellular staining and multicolor flow cytometry. Over a dose range of 10-100ng LPS/ml, TNF protein production was significantly higher in the CD43++ monocyte subset. At 1000ng LPS/ml 90% of all CD43++ monocytes stained positive for TNF and in terms of fluorescence intensity TNF was 5-fold higher compared to the CD43+ monocytes. These data indicate that the murine CD43++ monocyte subset exhibits features of pro-inflammatory monocytes and is functionally homologeous to the human CD14+CD16+ monocytes.

AB - Monocyte heterogeneity has been studied extensively in man but only recently tools have been developed to study blood monocyte populations in the mouse. We have used the MacGreen mouse model, which expresses the green fluorescent protein under the control of the promoter of the murine M-CSF receptor (CSF1 receptor, c-fms). Since both monocytes and granulocytes show GFP expression in this model the latter cells were excluded by staining with the Ly6G granulocyte marker. GFP+ Ly6G- blood monocytes were found to account for an average of 246+/-121cells/microl in these mice. These monocytes can be subdivided into CD43+ GR-1+ cells and CD43++ GR-1(-) cells, with the latter cells accounting for 140+/-77cells/mul, i.e. about 60% of all blood monocytes. After intraperitoneal injection of lipopolysaccharide (LPS) both blood monocyte subpopulations were depleted. The same was true after intranasal infection with Streptococcus pneumoniae but here the CD43++ subpopulation was preferentially reduced to 4cells/mul. For the study of TNF expression cells were stimulated in vitro with LPS from Salmonella abortus equi in the presence of Brefeldin A followed by intracellular staining and multicolor flow cytometry. Over a dose range of 10-100ng LPS/ml, TNF protein production was significantly higher in the CD43++ monocyte subset. At 1000ng LPS/ml 90% of all CD43++ monocytes stained positive for TNF and in terms of fluorescence intensity TNF was 5-fold higher compared to the CD43+ monocytes. These data indicate that the murine CD43++ monocyte subset exhibits features of pro-inflammatory monocytes and is functionally homologeous to the human CD14+CD16+ monocytes.

KW - Animals

KW - Leukosialin/genetics

KW - Lipopolysaccharides/pharmacology

KW - Mice

KW - Mice, Inbred C57BL

KW - Monocytes/drug effects

KW - Streptococcus pneumoniae/immunology

KW - Tumor Necrosis Factor-alpha/genetics

KW - Up-Regulation

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DO - 10.1016/j.imlet.2008.03.012

M3 - Article

VL - 118

SP - 142

EP - 147

JO - Immunology Letters

JF - Immunology Letters

SN - 0165-2478

IS - 2

ER -