The Facultative Intracellular PathogenCandida glabrataSubverts Macrophage Cytokine Production and Phagolysosome Maturation

In addition to its well-established effect on T cells, cyclosporin A (CsA) also inhibits inflammatory cytokine production by macrophages. However, little is known about the mechanism of action of CsA on macrophage cytokine production. We measured the effect of CsA on basal and phorbol-myristate-acetate (PMA)-stimulated production of interleukin-6 using the human monocyte cell line U937 differentiated with dimethylsulfoxide (DMSO). Interleukin-6 levels were measured in supernatant and cell lysates using specific enzyme-linked immunosorbent assays. We found that CsA decreases not only IL-6 release but also cytokine synthesis. The concentration of CsA used did not affect either cell viability or proliferation. Three possibilities may be advanced to explain the CsA-due decrease in IL-6 production by macrophages: (a) inhibition of the synthesis of an early common regulatory protein, (b) inhibition of cytokine gene transcription, or (c) modulation of post-transcriptional events. The first possibility was tested by measuring the effect of cycloheximide on the experimental system during the first 3 hours of culture. Although cycloheximide decreased total cytokine synthesis, the pattern of cytokine modulation by CsA persisted. These data suggest that CsA-mediated macrophage cytokine inhibition is not mediated by an early common regulatory protein. To further explore the inhibition mechanism, we measured IL-6 mRNA levels by Northern blot. IL-6 mRNA levels were unaffected by CsA both in resting and PMA-stimulated cells. We conclude that in human macrophages CsA diminishes IL-6 production at post-transcriptional level.

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Inhibition of primary murine macrophage cytokine production in vitro following treatment with the K-opioid agonist U50, 488H

Journal of Neuroimmunology ◽

10.1016/0165-5728(95)00159-x ◽

1996 ◽

Vol 64 (1) ◽

pp. 83-90 ◽

Cited By ~ 66

Author(s):

Candido Alicea ◽

Stanley Belkowski ◽

Toby K. Eisenstein ◽

Martin W. Adler ◽

Thomas J. Rogers

Keyword(s):

Cytokine Production ◽

Murine Macrophage ◽

Opioid Agonist ◽

Macrophage Cytokine

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Systemic Inflammation Induced by microRNAs: Endometriosis-Derived Alterations in Circulating microRNA 125b-5p and Let-7b-5p Regulate Macrophage Cytokine Production

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2017-01199 ◽

2017 ◽

Vol 103 (1) ◽

pp. 64-74 ◽

Cited By ~ 23

Author(s):

Sepide E Nematian ◽

Ramanaiah Mamillapalli ◽

Trisha S Kadakia ◽

Masoumeh Majidi Zolbin ◽

Sarah Moustafa ◽

...

Keyword(s):

Messenger Rna ◽

Cytokine Production ◽

University Hospital ◽

Regulatory Function ◽

Enzyme Linked Immunosorbent Assay ◽

Circulating Microrna ◽

Protein Levels ◽

Tnf Α ◽

Factor Α ◽

Macrophage Cytokine

Abstract Context Endometriosis is characterized by aberrant inflammation. We previously reported increased levels of microRNA (miRNA) 125b-5p and decreased levels of miRNA Let-7b-5p in serum of patients with endometriosis. Objective Determine the regulatory function of miRNAs 125b-5p and Let-7b-5p on production of proinflammatory cytokines in endometriosis. Design Case-control study. Setting University hospital. Patients Women with (20) and without (26) endometriosis; human U937 macrophage cell line. Intervention Sera were collected from surgically diagnosed patients and differentiated U937 cells that were transfected with miRNAs 125b-5p and Let-7b-5p mimics and inhibitor. Main Outcome Measures Enzyme-linked immunosorbent assay for tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-8, and IL-1β levels and quantitative real-time polymerase chain reaction for expression of miRNAs 125b-5p and Let-7b-5p in sera of patients with and without endometriosis. Transfected macrophages were evaluated for expression of inflammatory cytokines, intracellular production, and secretion of these cytokines. Results We noted substantial elevation of TNF-α, IL-1β, and IL-6, marked upregulation of miRNA 125b, and considerable downregulation of Let-7b in sera of patients with endometriosis vs control. There was a positive correlation between miRNA 125b levels and TNF-α, IL-1β, and IL-6 and a negative correlation between miRNA Let-7b levels and TNF-α in sera of patients with endometriosis. Transfection experiments showed a noteworthy upregulation of TNF-α, IL-1β, IL-6, and IL-8 in macrophages transfected with miRNA 125b mimic or Let-7b inhibitor. The secreted cytokine protein levels and intracellular imaging studies closely correlate with the messenger RNA changes. Conclusions Endometriosis-derived miRNAs regulate macrophage cytokine production that contributes to inflammation associated with this condition.

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