Abstract
Bacillus subtilis is a common group of probiotics that have been widely used in the feed industry as they can increase host resistance to pathogens and balance the immune response. However, the regulatory mechanism of Bacillus subtilis on the host immune system remains unclear in teleosts. In this study, we isolated and enriched dendritic cells from white blood cells (WBCs), and then stimulated them with Bacillus subtilis. Morphological features, specific biological functions, and authorized functional molecular markers were used in the identification of dendritic cells. Subsequently, we collected stimulated cells at 0, 4, and 18 h, and then constructed and sequenced the transcriptomic libraries. A transcriptome analysis showed that 2557 genes were up-regulated and 1708 were down-regulated at 4 h compared with the control group (|Fold Change| ≥ 4), and 1131 genes were up-regulated and 1769 were down-regulated between the cells collected at 18 h and 4 h (|Fold Change| ≥ 4). Gene Ontology (GO) annotations suggested many differentially expressed genes (DEGs) (p < 0.05 and |Fold Change| ≥ 4) were involved in immune-related biological functions including immune system progress, cytokine receptor binding, and cytokine binding. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the cytokine–cytokine receptor interaction pathways were significantly enriched at both time points (p < 0.05), which may play a key role in the response to stimulation. Furthermore, mRNA expression level examination of several pro-inflammatory cytokines and anti-inflammatory cytokines genes by quantitative real-time polymerase chain reaction (qRT-PCR) indicated that their expressions can be significantly increased in Bacillus subtili, which suggest that Bacillus subtilis can balance immune response and tolerance. This study provides dendritic cell (DC)-specific transcriptome data in grass carp by Bacillus subtilis stimulation, allowing us to illustrate the molecular mechanism of the DC-mediated immune response triggered by probiotics in grass carp.
Highlights
Probiotics are a kind of “live microorganism” that have a health benefit on their host [1]
The results of the flow forward/side scatter profile analysis indicate that this method can effectively enrich grass carp dendritic cells (Figure S1)
The results suggested that bacterial teichoic acids act as the key factor for inducing IL-10 production, which is mediated by TLR2-dependent ERK activation [52,53]
Summary
Probiotics are a kind of “live microorganism” that have a health benefit on their host [1]. Many studies have demonstrated that they can modulate the host homeostatic immune level and alleviate the inflammatory response caused by specific pathogens [2,3,4]. Dendritic cells (DCs) are the most efficient antigen-presenting cells (APCs) and play a central role in regulating the immune response and tolerance [7]. Many researchers have reported the effect of probiotics on the immune function of dendritic cells. The results suggested that DCs from UC patients have a different expression model in various immune-related molecules when compared with a control group. Due to the lack of specific molecule markers of DCs in teleosts, no studies have reported the modulation effects of probiotics on DCs in fish
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