Project description:To explore the regulatory network of noncoding RNAs after M.tb infection and the role of Rv1759c in the infection process, we collected samples of H37Rv- and H37Rv△1759c-infected macrophages and explored the full transcriptome expression profile. We constructed DE-lncRNA/DE-circRNA-DE-miRNA-DE-mRNA regulatory networks during H37Rv and H37Rv△1759c infection. In addition, we first discovered the close relationship between Rv1759c and chemokines during M.tb infection by comparing the transcription profiles of H37Rv and H37Rv△1759c and bioinformatics analysis. Here, our study comprehensively characterizes the ncRNA and mRNA profiles in macrophages infected with H37Rv and H37Rv△1759c, which provides support and new directions for in-depth exploration of ncRNA and PE/PPE family functions during the infection process.
Project description:The peritoneal macrophages were infected with Mtb H37Rv for 4 hours, and the miRNA expression profile were analyzed with deep sequencing.
Project description:In this study, RNA sequencing (Transcriptome sequencing) was employed to investigate the global transcriptome changes in the macrophages during the different strains of M.tb infection. THP-1 cells derived macrophages were exposed to the virulent M.tb strain H37Rv (Rv) or the avirulent M.tb strain H37Ra (Ra), and the M.tb BCG vaccine strain was used as a control. The cDNA libraries were prepared from M.tb infected macrophages and then sequenced.
Project description:Differentially regulated miRNA candidates in H37Rv infected THP-1 cells were analysed with respect to uninfected THP-1 reference samples. THP-1 cells are monocytes differentiated to macrophages after treatment with PMA for 48 hrs. Total RNA was isolated from infected THP-1 cells after 24 hrs of infection, cDNA was synthesized for TLDA real time PCR reaction using TaqMan MicroRNA Reverse Transcription kit and Megaplex Human Pool A and Pool B stem loop RT primers (version 3.0) as per manufacturer’s protocol. Further real time reaction was performed on QuantStudio 12K Flex Real-Time PCR System (Applied Biosystems) by using cDNA (without pre-amplification) on TLDA card A and card B (version 3.0).
Project description:Human monocyte-derived macrophages were infected with Mycobacterium tuberculosis (H37Rv strain) at a multiplicity of infection 1:1. After 2h of incubation, cells were washed to remove extracellular bacteria and were incubated in fresh medium for a further 18h in the presence or not of 100microM ATP. Total RNA was then extracted and gene expression was determined using HuGene 1.0 ST microarrays (Affymetrix, Inc.,Santa Clara, CA).
Project description:THP-1 cells were differentiated into macrophages with 0,1 µM PMA overnight then infected for 2 days with either H37Rv, M. marinum or heat killed H37Ra. Then RNA was collected in order to quantify the inflammatory response transcription as compared with the relative uninfected controls.
Project description:As macrophages are the primary site of Mtb infection and are sites of vitamin D signaling, we have used these cells to understand the molecular mechanisms underlying modulation of the immune response by the hormonal form of vitamin D, 1,25-dihydroxyvitamin D (1,25D). Microarrays were used to measure the changes in gene expression induced by 1,25D treatment of H37Rv-infected THP-1 cells