Skip to main content
Dryad logo

DNA microarray of long-lived neutrophils versus control

Citation

Poubelle, Patrice (2021), DNA microarray of long-lived neutrophils versus control, Dryad, Dataset, https://doi.org/10.5061/dryad.sf7m0cg56

Abstract

The concept of plasticity of neutrophils is highlighted by studies showing their ability to transdifferentiate into antigen-presenting cells. In this regard, transdifferentiated neutrophils were found at inflammatory sites of autoimmune arthritis (AIA). Exposure of neutrophils to inflammatory stimuli prolongs their survival, thereby favoring the acquisition of pathophysiologically relevant phenotypes and functions. By using microarrays, qRT-PCR, and ELISAs, we showed that long-lived (LL) neutrophils obtained after 48 h of culture in the presence of GM-CSF, TNF, and IL-4 differentially expressed genes related to apoptosis, MHC class II, immune response, and inflammation. The expression of anti-inflammatory genes mainly of peptidase inhibitor families is upregulated in LL neutrophils. Among these, the PI3 gene encoding elafin was the most highly expressed. The de novo production of elafin by LL neutrophils depended on a synergism between GM-CSF and TNF via the activation and cooperativity of CCAAT/enhancer-binding protein ß and NF-κB pathways, respectively. Elafin concentrations were higher in synovial fluids (SF) of patients with AIA than in SF of osteoarthritis. SF neutrophils produced more elafin than blood counterparts. These results are discussed with respect to implications of neutrophils in chronic inflammation and the potential influence of elafin in AIA.

Methods

DNA microarray analyses were carried out with Affymetrix Human Transcriptome Array 2.0 (HTA 2.0) according to the Affymetrix standard protocol.

To validate the results of our microarray data, the expression of selected genes in long-lived neutrophils were analyzed by qRT-PCR according to MIAME guidelines

Usage Notes

This analysis allowed generating a gene list with the upregulated and downregulated genes in LL neutrophils with a fold change of at least 2, with a p value < 0,01 and an FDR value < 0,1.

Funding

Canadian Institutes of Health Research