Project description:The emergence of multidrug resistant (MDR) Mycobacterium tuberculosis (Mtb) strains, resistant to the frontline anti-tubercular drugs rifampicin and isoniazid, forces treatment with less effective and toxic second-line drugs and stands to derail TB control efforts. However, the immune response to MDR Mtb infection remains poorly understood. Here, we determined the RNA transcriptional profile of in vitro generated macrophages to infection with either drug susceptible Mtb HN878 or MDR Mtb W_7642 infection.
Project description:Non-typhoidal Salmonella (NTS) are among of the most important food-borne pathogens. Recently, a highly invasive multi-drug resistant S. Typhimurium of a distinct multilocus sequence type (MLST), ST313, has emerged across sub-Saharan Africa as a major cause of lethal bacteraemia in children and immunosuppressed adults. Encounters between dendritic cells (DCs) and invading bacteria determine the course of infection but whether or how ST313 might usurp DC mediated defence has not been reported. Here we utilised fluorescently labelled invasive and non-invasive strains of Salmonella combined with single-cell RNA sequencing to study the transcriptomes of individual infected and bystander DCs. The transcriptomes displayed a repertoire of cell instrinsic and extrinsic innate response states that differed between invasive and non-invasive strains. Gene expression heterogeneity was increased in DCs challenged with invasive Salmonella. DCs exposed but not harbouring invasive Salmonella exhibited a hyper-activated profile that likely facilitates trafficking of infected cells and dissemination of internalised intact bacteria. In contrast, invasive Salmonella containing DCs demonstrate reprogramming of trafficking genes required to avoid autophagic destruction. Furthermore, these cells displayed differential expression of tolerogenic IL10 and MARCH1 enabling CD83 mediated adaptive immune evasion. Altogether our data illustrate pathogen cell-to cell variability directed by a Salmonella invasive strain highlighting potential mechanisms of host adaption with implications for dissemination in vivo.
Project description:Non-typhoidal Salmonella (NTS) are among of the most important food-borne pathogens. Recently, a highly invasive multi-drug resistant S. Typhimurium of a distinct multilocus sequence type (MLST), ST313, has emerged across sub-Saharan Africa as a major cause of lethal bacteraemia in children and immunosuppressed adults. Encounters between dendritic cells (DCs) and invading bacteria determine the course of infection but whether or how ST313 might usurp DC mediated defence has not been reported. Here we utilised fluorescently labelled invasive and non-invasive strains of Salmonella combined with single-cell RNA sequencing to study the transcriptomes of individual infected and bystander DCs. The transcriptomes displayed a repertoire of cell instrinsic and extrinsic innate response states that differed between invasive and non-invasive strains. Gene expression heterogeneity was increased in DCs challenged with invasive Salmonella. DCs exposed but not harbouring invasive Salmonella exhibited a hyper-activated profile that likely facilitates trafficking of infected cells and dissemination of internalised intact bacteria. In contrast, invasive Salmonella containing DCs demonstrate reprogramming of trafficking genes required to avoid autophagic destruction. Furthermore, these cells displayed differential expression of tolerogenic IL10 and MARCH1 enabling CD83 mediated adaptive immune evasion. Altogether our data illustrate pathogen cell-to cell variability directed by a Salmonella invasive strain highlighting potential mechanisms of host adaption with implications for dissemination in vivo.
Project description:To gain insight into the alterations of gene expression profile in the course of non-mutationally acquired resistance, we performed RNA-seq comparing MDR persister cells to MDR cancer cells.
Project description:Septoria leaf blotch is a worldwide threat for wheat and mainly controlled by the application of synthetic fungicides. The fungal pathogen responsible for this disease, Zymoseptoria tritici, was shown as highly adaptable to its host plant, but also to fungicide challenge. Over the past decades it developed resistance to most fungicides due to target site modifications. Recently isolated strains showed cross-resistance to diverse fungicides and to unrelated drugs, suggesting a resistance mechanism that seems rarer in phytopathogenic fungi, known as multidrug resistance (MDR) in other organisms. In this study we show for two Z. tritici MDR strains, MDR6 and MDR7, enhanced prochloraz efflux sensitive to the modulators amitryptiline and chlorpromazine. Efflux was also inhibited by verapamil in the MDR7strain. Transcriptomics revealed several overexpressed transporter genes in both MDR strains, out of which the expression of the MgMFS1 transporter gene was the strongest and constitutively high in tested MDR field strains. Its inactivation in the MDR6 strain abolished resistance to fungicides with different modes of action revealing its involvement in the MDR phenomenon in Z. tritici.
Project description:Up-regulation of the neuropeptide NTS in a subgroup of lung cancers has been linked to poor prognosis. However, the regulatory pathway centered on NTS in lung cancer remains unclear. Here we identified the NTS specific enhancer in lung adenocarcinoma cells. The AF4/FMR2 (AFF) family protein AFF1 occupies the NTS enhancer and inhibits NTS transcription. Clustering analysis of lung adenocarcinoma gene expression data demonstrated that NTS is highly positively correlated with the expression of the oncogenic factor CPS1. Detailed analyses demonstrated that NTS antagonizes the IL6 pathway in regulating CPS1. Thus, our analyses revealed a novel NTS centered regulatory axis, consisting of AFF1 as a master transcription suppressor and IL6 as an antagonist in lung adenocarcinoma cells.
Project description:We examined the microRNA profiles of THP-1 macrophages upon the MTB infection of (1) Beijing/W and non-Beijing/W clinical strains, and (2) susceptible and multidrug-resistant (MDR-) MTB strains.
Project description:Septoria leaf blotch is a worldwide threat for wheat and mainly controlled by the application of synthetic fungicides. The fungal pathogen responsible for this disease, Zymoseptoria tritici, was shown as highly adaptable to its host plant, but also to fungicide challenge. Over the past decades it developed resistance to most fungicides due to target site modifications. Recently isolated strains showed cross-resistance to diverse fungicides and to unrelated drugs, suggesting a resistance mechanism that seems rarer in phytopathogenic fungi, known as multidrug resistance (MDR) in other organisms. In this study we show for two Z. tritici MDR strains, MDR6 and MDR7, enhanced prochloraz efflux sensitive to the modulators amitryptiline and chlorpromazine. Efflux was also inhibited by verapamil in the MDR7strain. Transcriptomics revealed several overexpressed transporter genes in both MDR strains, out of which the expression of the MgMFS1 transporter gene was the strongest and constitutively high in tested MDR field strains. Its inactivation in the MDR6 strain abolished resistance to fungicides with different modes of action revealing its involvement in the MDR phenomenon in Z. tritici. A total of four strains were compared, two sensitive (IPO323, S6) and two MDR strains (09-ASA-3apz; 09-CB01) with three replicates each. All strains were grown in liquid YPD medium to exponential growth.
Project description:In this study, we have defined the NsrR regulon in Salmonella enterica sv. Typhimurium 14028s using a transcriptional microarray. Wild-type and nsrR mutant S. Typhimurium were grown aerobically to early log-phase (OD600~0.5) at 37C in LB medium. Total RNA was isolated from three independent cultures of both strains and interrogated on a PCR product array representing almost all ORFs.