Comparative analysis using different multiplicity of infection ratios from Chlamydia-infected epithelial cells
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ABSTRACT: Here we examined gene expression from an in-vitro dual-RNA-seq experiment using Chlamydia trachomatis-infected HEp-2 epithelial cells. The experimental design consisted of three different multiplicity of infection ratio's (0.1, 1 and 10) across two different timeframes (1 and 24hrs), in addition to applying two different depletion methods (rRNA and rRNA + PolyA depletion). The aim was to examine and outline which MOI and depletion methods are most suited for bacterial-based infections.
Project description:Here we examined host cell gene expression in response to Chlamydia trachomatis infection by applying scRNA-Seq to in vitro C. trachomatis-infected HEp-2 epithelial cells and time-matched uninfected cells over the early chlamydial developmental cycle (3, 6 and 12 hours). We collected 264 single cells across both conditions all time points. The aims of the experiment were examining host cell responses to infection at single cell resolution, and identifying early host cell signatures of infection.
Project description:In this project we examined the in-vitro effect of female sex hormones (estradiol and progesterone at average physiological concentrations) during a infection mediated by Chlamydia trachomatis serovar D, on the gene expression of human endometrial cell line ECC-1 The effects of the female sex hormones progesterone and oestradiol while infected by Chlamydia trachomatis were examined at two timepoints.
Project description:Chlamydia trachomatis are the etiological agents of a range of diseases and are epidemiologically associated with cervical and ovarian cancers. The interplay between host and chlamydia is highly complex, and to obtain panoramic view of the functional interplay, we performed combinatorial global phosphoproteomic and transcriptomic analyses of C. trachomatis-induced signaling. We identified numerous previously unknown C. trachomatis phosphoproteins and C. trachomatis-regulated host phosphoproteins that are substrates of kinases involved in various cellular processes. Interestingly, several host transcription factors (TFs) that are phosphorylated in C. trachomatis infections, including ETS2 repressor factor (ERF), proto-oncogenic transcription factor ETS1 are targets of ERK MAPK signaling. While these TFs were found to be essential for Chlamydia development, we demonstrated their involvement in inducing epithelial-to-mesenchymal transition in C. trachomatis infected cells by transcriptional regulation of genes involved in cellular motility and invasion. Our data reveals substantially unexplored complexity of C. trachomatis-induced signaling and provides broader insights into pro-carcinogenic potential of C. trachomatis.
Project description:Neutrophil granulocytes are the major cells involved in the Chlamydia trachomatis (C.trachomatis)-mediated inflammation and histopathology. A key gene in human intracellular antichlamydial defense is the tryptophan degrading enzyme indoleamine 2,3-dioxygenase (IDO), which limits the growth of the tryptophan auxotroph Chlamydia. Despite its importance, the role of IDO in the intracellular defense against Chlamydia in neutrophils has not yet been characterized. Affymetrix microarrays were used to obtain global gene expression data for monitoring the effect of C. trachomatis serovar D infection on the transcriptome of human neutrophil granulocytes.
Project description:Chlamydia trachomatis D serovar was grown in axenic culture with G6P or G6P with glutamine. The data reveal the early transcriptonal regulation in the bacteria.
Project description:Chlamydia trachomatis is an obligate intracellular pathogen that causes trachoma and sextually transmitted disease in human. During early stage of infection, Chlamydia secreted bacterial effector proteins into host cell cytoplasm to help its entry and estabilishment of early replicated niche. We identified a Chlamydia mutant that lack an early Effector. To address the function of this effector, we infected A2EN cells with this mutant (G1V) and its complemented counterpart (G1TEPP) to see what host gene transcriptions are affected by this effector. A2EN cells were mock infected, or infected with a Chlamydia mutant or its complemented counterpart for 4 hour post infection.