Supplementary Materialsijms-20-06207-s001. might control immune activity in macrophages. This study provides potential insight into the functions of lncRNA in macrophages regulated by IL-27. Value= 3). Data were normalized to the house keeping gene GAPDH. The X-axis is usually LHRI_LNC-1, 2, 3, 4, 5, 6, 7, 8; and the Y-axis is usually relative gene expression. 2.7. Biological Function Assay We have previously found Rabbit Polyclonal to NCR3 that IL-27 inhibits HIV in macrophages and T cells [25,41,42] and as shown in this study, IL-27 modulates lncRNA profiles. To assess the role of several novel lncRNAs as a pilot experiment, we randomly selected four lncRNAs, LHRI_LNC-2, 3, 5, and 6, and made an expression vector. HEK293T cells were transfected with different concentrations (10, 100, and 1000 ng) of each plasmid, followed by HIVluc contamination. HIV contamination was monitored by luciferase activity as described in the materials and methods. Our results indicated that this selected lncRNAs had no significant impact on HIV contamination (Physique S5). 3. Discussion In the present study, we analyzed the mRNA (coding gene) and lncRNA expression profiles and functional networks of IL-27 treated macrophages. We identified 434 protein-coding genes and 146 lncRNAs (including 88 unknown ones) with differential expression in both the ABI and IMAC experiments. Both the KEGG pathway and GO-term Cephapirin Sodium enrichment analysis indicated that some cellular functional pathways contribute to IL-27 functions such as immune response, inflammatory response, antigen processing and presentation, and cell cycle. Further co-expression network analysis revealed that some lncRNAs may be involved in the biological functions through their target genes. Therefore, our results provide a comprehensive profile of the lncRNA and mRNA expression in the macrophage regulated by IL-27 and lay a foundation for the understanding of the IL-27 immunoregulatory mechanisms. Macrophages are one of the antigen-presenting cells in the immune system. They modulate many pathophysiological pathways and are activated by pathogens. The role of lncRNAs in macrophage activation has been revealed by several studies [14,43]. Scacalossi et al. recently reviewed the Cephapirin Sodium role of lncRNA in macrophage function such as in macrophage polarization, and innate and adaptive immune functions . Gupta et al. analyzed lncRNAs in bovine macrophages and suggested that lncRNAs could respond to para-tuberculosis contamination . Our results showed that lncRNAs regulated by IL-27 in human macrophages were mainly involved in immune response and cell cycle pathways. Vadiveloo reported that cell cycle proteins play a role in macrophage activation . We previously reported that IL-27 inhibits HIV in macrophages . Other experts have also reported that IL-27 is able to suppress HBV, HCV, HIV-1, HIV-2, HSV, and influenza A [30,31,32,33,34] in various cell types. In this study, KEGG analysis indicated that this differential profile of the expressed gene was enriched in immune diseases such as staphylococcus aureus contamination, autoimmune thyroid disease, inflammatory bowel disease (IBD), and graft-versus-host disease. It has been reported that IL-27 is usually elevated in patients with autoimmune/inflammatory diseases such as Crohns disease, rheumatoid arthritis, multiple sclerosis, psoriasis, and aplastic anemia . Mutations in IL-27 could be a causal effect for some autoimmune diseases such as IBD, chronic obstructive pulmonary disease, and asthma . Cephapirin Sodium The Gene Ontology (GO) knowledgebase analysis exhibited that differentially expressed genes were enriched in immune related GO terms such as interferon-gamma (IFN-)-mediated signaling pathway, T cell co-stimulation, antigen processing and presentation, and the major histocompatibility complex (MHC) class II protein complex. Rajaiah et.