Transcriptome analysis of porcine PBMCs reveals lipopolysaccharide-induced immunomodulatory responses and crosstalk of immune and glucocorticoid receptor signaling

Abstract

The current level of knowledge on transcriptome responses triggered by endotoxins and glucocorticoids in immune cells in pigs is limited. Therefore, in the present study, we treated porcine peripheral blood mononuclear cells (PBMCs) with lipopolysaccharide (LPS) and dexamethasone (DEX) separately or combined for 2 hours. The resultant transcriptional responses were examined by mRNA sequencing. We found that the LPS treatment triggered pronounced inflammatory responses as evidenced by upregulation of pro-inflammatory cytokines, chemokines, and related signaling pathways like NF-κB. Concurrently, a series of downregulated pro-inflammatory and upregulated anti-inflammatory molecules were identified. These are involved in the inhibition of TLR, NF-κB, and MAPK cascades and activation of signaling mediated by Tregs and STAT3, respectively. These findings suggested that LPS initiated also an anti-inflammatory process to prevent an overwhelming inflammatory response. The transcriptome responses further revealed substantial crosstalk of immune responses and glucocorticoid receptor (GR) signaling. This was apparent in four aspects: constitutive inhibition of T cell signaling by DEX through a subset of genes showing no response to LPS; inhibition of LPS-induced inflammatory genes by DEX; attenuation of DEX action by LPS paralleled by the regulation of genes implicated in cytokine and calcium signaling; and DEX-induced changes in genes associated with the activation of proinflammatory TLR, NF-κB, iNOS, and IL-1 signaling. Consequently, our study provides novel insights into inflammatory and GR signaling in pigs, as well as an understanding of the application of glucocorticoid drugs for the treatment of inflammatory disorders.

miR-642a-5p increases glucocorticoid sensitivity by suppressing the TLR4 signalling pathway in THP-1 cells

“…In rheumatic diseases, TLR4 gene polymorphisms are closely related to the GC response [ 41 ]. Many studies have shown that NF-κB activation through various pathways can inhibit the production of anti-inflammatory factors by preventing GR from binding specific DNA regions of the GRE, the first step in the GC response, leading to GC resistance [ [42] , [43] , [44] ]. Our study also found miR-642a-5p mimic enhanced the inhibitory effect of DEX on TLR4, promoted GR translocation into the nucleus, and decreased the levels of LPS-induced inflammatory factors.…”

Glucocorticoid receptor hypersensitivity enhances inflammatory signaling and inhibits cell cycle progression in porcine PBMCs

“…The mRNA sequencing data presented and analyzed in the current report were first published and all the sampling and treatment procedures were done in our previous study ( 20 ). Briefly, PBMCs were obtained from 24 purebred German Landrace pigs (≈170 days old) with equal numbers of males and females and balanced for the three GR genotypes (Ala/Ala, Ala/Val, and Val/Val).…”

“…PBMCs from each animal were distributed into four treatment groups and treated for 2 h at 37°C in a 5% CO 2 atmosphere with: vehicle (CON group); DEX (5 nM); LPS (10 μg/mL); and LPS + DEX, respectively. The mRNA sequencing was performed using the Illumina HiSeq 2500 platform at the FBN sequencing facility as described previously ( 20 ). The resultant sequencing data were first submitted to the ArrayExpress repository ( ) with accession number E-MTAB-9808, as outlined in our earlier study ( 20 ).…”

“…The mRNA sequencing was performed using the Illumina HiSeq 2500 platform at the FBN sequencing facility as described previously ( 20 ). The resultant sequencing data were first submitted to the ArrayExpress repository ( ) with accession number E-MTAB-9808, as outlined in our earlier study ( 20 ).…”

Glucocorticoid-induced Changes in the Transcriptional Activity of Genes of the Innate and Adaptive Immune System in the Blood of Patients with Acute Urticaria

“…The GCs receptors (GRs) directly bind with nuclear factor kappa B (NF-κB) and AP-1 family proteins to inhibit their transcriptional activity [28]. The outcome is the following: GCs suppress T-cell expression of costimulatory molecules (such as CD28), cytokines (IL-2, IL-4, IL-5, IL-6, IL-8, TNF, and IFNγ), and chemokines (CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, and CCL13) [29]. Furthermore, GCs upregulate coinhibitory molecules such as PD1, CTLA4, LAG3, and TIM3 [30].…”