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Schieck, E.

Publications and source records attributed to Schieck, E..

2 recordsLinked to original sources

Host transcriptional programs underlying lesion development in contagious bovine pleuropneumonia

Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides (Mmm), remains a major burden to cattle health and the agricultural industry. Mmm is an atypical bacterial pathogen that appears to lack classical virulence factors that cause direct tissue injury (i.e. toxins), and little is known about the mechanisms driving its pathogenicity. The host immune response is believed to be implicated in CBPP pathology, though the molecular mechanisms underlying lesion initiation, progression and chronicity are poorly defined. Classical pathology describes a continuum of lung lesions starting from early inflammation to more mature necrotic lesions and formation of fibrotic sequestra. However, the host transcriptional response driving this potentially immunopathological progression during Mmm infection has never been resolved in vivo. Here, we performed lesion-stage-resolved transcriptomic profiling of pathological lung tissue collected from experimentally infected animals and compared to healthy lung tissue collected from unchallenged controls. Differential gene expression and functional enrichment analyses were used to identify biological pathways relevant to Mmm infection and pathological lesion formation. Early infection was dominated by interferon-stimulated genes and cytokine-responsive pathways, creating a primarily antiviral-like response environment despite the bacterial etiology. Red hepatization showed strong induction of neutrophil chemoattractants, epithelial remodeling markers, and early matrix-remodeling enzymes. Consolidation, spanning red and grey stages, was enriched for innate immune activation, leukocyte adhesion, extracellular matrix organization, and persistent interferon signaling. Grey hepatization reflected late-stage consolidation with heightened neutrophil effector activity, oxidative and proteolytic injury, and macrophage and fibroblast-linked collagen processing. Necrosis/Sequestra lesions showed reduced inflammatory signaling, robust extracellular matrix organization, adhesion, and morphogenetic pathways consistent with encapsulation and sequestrum formation. Our data indicate that the dynamic continuum of CBPP lung pathology is initiated by interferon-primed myeloid recruitment and amplified by neutrophil-driven injury and macrophage- and fibroblast-mediated matrix remodeling. These data further substantiate the role of dysregulated immunity in the development of disease during Mmm infection.

immunology↗

Removal of a subset of non-essential genes fully attenuates a highly virulent Mycoplasma strain

Mycoplasmas are the smallest free-living organisms and cause a number of economically important diseases affecting humans, animals, insects and plants. Here, we demonstrate that highly virulent Mycoplasma mycoides subspecies capri (Mmc) can be fully attenuated via targeted deletion of non-essential genes encoding, among others, potential virulence traits. Five genomic regions, representing approximately ten percent of the original Mmc genome, were successively deleted using Saccharomyces cerevisiae as an engineering platform. Specifically, a total of 68 genes out of the 432 genes verified to be individually nonessential in the JCVI-Syn3.0 minimal cell, were excised from the genome. In vitro characterization showed that this mutant was similar to its parental strain in terms of its doubling time, even though ten percent of the genome content were removed. A novel in vivo challenge model in goats revealed that the wild-type parental strain caused marked necrotizing inflammation at the site of inoculation, septicemia and all animals reaching endpoint criteria within seven days after experimental infection. This is in contrast to the mutant strain, which caused no clinical signs nor pathomorphological lesions. These results highlight, for the first time, the rational design, construction and complete attenuation of a Mycoplasma strain via synthetic genomics tools. Trait addition using the yeast-based genome engineering platform and subsequent in vitro or in vivo trials employing the Mycoplasma chassis will allow us to dissect the role of individual candidate Mycoplasma virulence factors and lead the way for the development of an attenuated designer vaccine.\n\nIMPORTANCEMembers of the Mycoplasma mycoides cluster cause important animal plaques in Africa and Asia, which impact animal welfare, provision of food and the life of thousands of small-scale farmers. We applied synthetic biology tools to Mycoplasma mycoides in order to design and create a fully attenuated Mycoplasma strain that was subsequently confirmed in vivo using a novel caprine infection model. This is the first time that a Mycoplasma mutant developed by applying synthetic biology tools has been tested in vivo to pin point candidate virulence traits. The mutant strain is similar to \"apathogenic E. coli K12\" strains that boosted the research on host-pathogen interactions for the genus Escherichia and other bacterial genera.

microbiology↗