Purification of human blood monocytes by hypotonic density gradient centrifugation in Percoll

Feige, Ulrich; Overwien, Bruno; Sorg, Clemens

doi:10.1016/0022-1759(82)90315-5

Cited by 70 publications

(27 citation statements)

References 22 publications

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“…Peripheral blood granulocytes and monocytes were isolated from buffy coats using Ficoll-Paque gradient centrifugation (26). Erythrocytes were removed from sedimented granulocytes through hypotonic lysis.…”

Section: Isolation Of Human Pblsmentioning

confidence: 99%

An Annexin 1 N-Terminal Peptide Activates Leukocytes by Triggering Different Members of the Formyl Peptide Receptor Family

Ernst¹,

Lange²,

Wilbers³

et al. 2004

The Journal of Immunology

146

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The human N-formyl peptide receptor (FPR) is a key modulator of chemotaxis directing granulocytes toward sites of bacterial infections. FPR is the founding member of a subfamily of G protein-coupled receptors thought to function in inflammatory processes. The other two members, FPR-like (FPRL)1 and FPRL2, have a greatly reduced affinity for bacterial peptides or do not bind them at all, with FPRL2 being considered an orphan receptor so far. In this study we show that a peptide derived from the N-terminal domain of the anti-inflammatory protein annexin 1 (lipocortin 1) can activate all three FPR family members at similar concentrations. The annexin 1 peptide initiates chemotactic responses in human monocytes that express all three FPR family members and also desensitizes the cells toward subsequent stimulation with bacterial peptide agonists. Experiments using HEK 293 cells stably expressing a single FPR family member reveal that all three receptors can be activated and desensitized by the N-terminal annexin 1 peptide. These observations identify the annexin 1 peptide as the first endogenous ligand of FPRL2 and indicate that annexin 1 participates in regulating leukocyte emigration into inflamed tissue by activating and desensitizing different receptors of the FPR family.

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Section: Isolation Of Human Pblsmentioning

confidence: 99%

An Annexin 1 N-Terminal Peptide Activates Leukocytes by Triggering Different Members of the Formyl Peptide Receptor Family

Ernst¹,

Lange²,

Wilbers³

et al. 2004

The Journal of Immunology

146

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“…Human monocytes were isolated from human buffy coats by Ficoll-Paque (Pharmacia, Freiburg, Germany) and subsequent Percoll (Pharmacia) density gradient centrifugation (monocyte purity > 90 %) [20]. Monocytes (2 x lo6 cells/ml) were cultured in hydrophobic Teflon bags (Heraeus, Hanau, Germany) in McCoy's 5a medium supplemented with 20 % FCS at 37 "C in 5 YO C 0 2 in air for 18 h.…”

Section: Isolation Of Blood Mononuclear Cellsmentioning

confidence: 99%

Identification of a novel surface molecule, RM3/1, that contributes to the adhesion of glucocorticoid‐induced human monocytes to endothelial cells

1996

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“…Uppsala, Swe den) followed by density gradient centrifugation in Percoll [Boyum. 1968: Feige et al. 1982Zwadlo et al, 1986].…”

Section: Cellsmentioning

confidence: 99%

Characterization and Expression Kinetics of an Endothelial Cell Activation Antigen Present in vivo Only in Acute Inflammatory Tissues

Goerdt¹,

Zwadlo²,

Schlegel³

et al. 1987

Pathobiology

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Endothelial cell activation by endotoxin (LPS), tumor necrosis factor (TNF), Interleukin-1-α, β (IL-1-α β) and phorbolesters (TPA) results in increased monocyte adhesion. Examination of kinetics of monocyte adhesion shows that the onset of adherence enhancement (AE) is similar in all five agents (about 300% AE at 6 h), while its decrease is delayed in LPS/TNF versus IL-1-α, β /TPA-induced activation (LPS versus IL-1- β:260% versus 60% at 18 h). Monoclonal antibody (4D10), raised against 24 h LPS-stimulated endothelial cells detects an endothelial cell-specific activation antigen at M_r 81,000 that is induced by LPS, TNF, IL-1-α β and TPA (within 6 h about 100% positive cells). Decrease in antigen-positive cells is delayed in LPS/TNF versus IL-1-α, β /TPA-induced antigen expression (LPS vs. IL-1-β: 60% vs. 5% at 24 h). In situ the antigen is not expressed in normal and chronic inflammatory tissues. Acute inflammatory tissues, including contact and atopic dermatitis, psoriasis and periodontitis, however, show endothelial cells staining strongly positive. In contact eczemas at different times after elicitation (0, 6, 24, 72, 96 h), expression of the antigen is first seen after 24 h and is still strong at 96 h. These data indicate that LPS/TNF conduct an endothelial cell activation program in vitro, showing the same prolonged kinetics that is found for endothelial cell activation in the acute inflammatory process in vivo.

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Purification of human blood monocytes by hypotonic density gradient centrifugation in Percoll

Cited by 70 publications

References 22 publications

An Annexin 1 N-Terminal Peptide Activates Leukocytes by Triggering Different Members of the Formyl Peptide Receptor Family

An Annexin 1 N-Terminal Peptide Activates Leukocytes by Triggering Different Members of the Formyl Peptide Receptor Family

Identification of a novel surface molecule, RM3/1, that contributes to the adhesion of glucocorticoid‐induced human monocytes to endothelial cells

Characterization and Expression Kinetics of an Endothelial Cell Activation Antigen Present in vivo Only in Acute Inflammatory Tissues

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