A simple method for the purification of human peripheral blood monocytes

Denholm, Elizabeth M.; Wolber, Frances M.

doi:10.1016/0022-1759(91)90092-t

Cited by 76 publications

(44 citation statements)

References 8 publications

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“…Monocytes were first enriched by centrifugation over a Percoll density gradient as described (38) and purified by cell-sorting procedure as described (28) (Epics Elite ESP; Coulter Electronics Canada, Burlington, Ontario, Canada). This procedure yielded Ͼ98% pure monocyte suspensions as determined by CD14 staining.…”

Section: Methodsmentioning

confidence: 99%

Activation of Monocyte Cyclooxygenase-2 Gene Expression by Human Herpesvirus 6

Janelle¹,

Gravel²,

Gosselin³

et al. 2002

Journal of Biological Chemistry

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Prostaglandin E 2 (PGE 2 ) is an arachidonic acid metabolite mainly produced by activated monocytes/macrophages (Mo/M) that display broad immunomodulatory activities. Several viruses capable of infecting Mo/M modulate PGE 2 synthesis in a way that favors the infection processes and the spread of virions. In the present work, we studied the effect of human herpesvirus 6 (HHV-6) infection of Mo/M on PGE 2 synthesis. Our results indicate that HHV-6 induces COX-2 gene expression and PGE 2 synthesis within a few hours of infection. We mapped the different promoter elements associated with COX-2 gene activation by HHV-6 to two cis-acting elements: a cyclic AMP-responsive element and an activator protein-1 element. HHV-6 immediate-early protein 2 was identified as a modulator of COX-2 gene expression in Mo/M. Finally, addition of PGE 2 to HHV-6-infected peripheral blood mononuclear cells cultures was found to increase significantly viral replication. Overall, these results further contribute to the immunomodulatory properties of HHV-6 and highlight a potential role for eicosanoids in the replication process of this virus.

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Section: Methodsmentioning

confidence: 99%

Activation of Monocyte Cyclooxygenase-2 Gene Expression by Human Herpesvirus 6

Janelle¹,

Gravel²,

Gosselin³

et al. 2002

Journal of Biological Chemistry

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“…Monocytes destined for use in the assessment of chemotaxis were obtained by centrifugation on a Percoll gradient [16]. Following the washing steps to remove the platelets (see above), the mixed mononuclear ceils were resuspended in 4 ml of PBS and mixed with 8 ml Percoll:Hank's balanced salt solution (6:1, pH 7.0) and spun for 30 min at 400xg at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Cytotoxic and chemotactic potencies of several aldehydic components of oxidised low density lipoprotein for human monocyte‐macrophages

et al. 1996

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“…To prepare PBL, PBMC were added to a continuous Percoll (Pharmacia, Piscataway, NJ) density gradient and the highdensity fraction was collected (9). T cells were prepared by immediate rosetting with 2-aminoethylisothiouronium bromide-treated SRBC (10).…”

Section: Lymphocyte Isolationmentioning

confidence: 99%

Generation Ex Vivo of TGF-β-Producing Regulatory T Cells from CD4+CD25− Precursors

Zheng

Gray

Ohtsuka

et al. 2002

The Journal of Immunology

411

316

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Previously we reported that TGF-β has an important role in the generation and expansion of human “professional” CD4+CD25+ regulatory T cells in the periphery that have a cytokine-independent mechanism of action. In this study we used low-dose staphylococcal enterotoxin to induce T cell-dependent Ab production. We report that TGF-β induces activated CD4+CD25− T cells to become Th3 suppressor cells. While stimulating CD4+ cells with TGF-β modestly increased expression of CD25 and intracellular CTLA-4 in primary cultures, upon secondary stimulation without TGF-β the total number and those expressing these markers dramatically increased. This expansion was due to both increased proliferation and protection of these cells from activation-induced apoptosis. Moreover, adding as few as 1% of these TGF-β-primed CD4+ T cells to fresh CD4+ cells and B cells markedly suppressed IgG production. The inhibitory effect was mediated by TGF-β and was also partially contact dependent. Increased TGF-β production was associated with a decreased production of IFN-γ and IL-10. Depletion studies revealed that the precursors of these TGF-β-producing CD4+ suppressor cells were CD25 negative. These studies provide evidence that CD4+CD25+ regulatory cells in human blood consist of at least two subsets that have TGF-β-dependent and independent mechanisms of action. TGF-β has an essential role in the generation of both of these T suppressor cell subsets from peripheral T cells. The ability to induce CD4+ and CD8+ cells to become regulatory cells ex vivo has the potential to be useful in the treatment of autoimmune diseases and to prevent transplant rejection.

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A simple method for the purification of human peripheral blood monocytes

Cited by 76 publications

References 8 publications

Activation of Monocyte Cyclooxygenase-2 Gene Expression by Human Herpesvirus 6

Activation of Monocyte Cyclooxygenase-2 Gene Expression by Human Herpesvirus 6

Cytotoxic and chemotactic potencies of several aldehydic components of oxidised low density lipoprotein for human monocyte‐macrophages

Generation Ex Vivo of TGF-β-Producing Regulatory T Cells from CD4+CD25− Precursors

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