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The Clostridium difficile Exosporium Cysteine (CdeC)-Rich Protein Is Required for Exosporium Morphogenesis and Coat Assembly
Clostridium difficile is an important nosocomial pathogen that has become a major cause of antibiotic-associated diarrhea.
There is a general consensus that C. difficile spores play an important role in C. difficile pathogenesis, contributing to infection,
persistence, and transmission. Evidence has demonstrated that C. difficile spores have an outermost layer, termed the exosporium,
that plays some role in adherence to intestinal epithelial cells. Recently, the protein encoded by CD1067 was shown to be
present in trypsin-exosporium extracts of C. difficile 630 spores. In this study, we renamed the CD1067 protein Clostridium difficile
exosporium cysteine-rich protein (CdeC) and characterized its role in the structure and properties of C. difficile spores.
CdeC is expressed under sporulation conditions and localizes to the C. difficile spore. Through the construction of an ÎcdeC
isogenic knockout mutant derivative of C. difficile strain R20291, we demonstrated that (i) the distinctive nap layer is largely
missing in ÎcdeC spores; (ii) CdeC is localized in the exosporium-like layer and is accessible to IgGs; (iii) Î cdeC spores were
more sensitive to lysozyme, ethanol, and heat treatment than wild-type spores; and (iv) despite the almost complete absence of
the exosporium layer, Î cdeC spores adhered at higher levels than wild-type spores to intestinal epithelium cell lines (i.e., HT-29
and Caco-2 cells). Collectively, these results indicate that CdeC is essential for exosporium morphogenesis and the correct assembly
of the spore coat of C. difficile.Keywords: Spore surface,
Subtilis,
Infection,
Germination,
Culture,
Gene,
Caco-2,
Bacterial spores,
Identification,
Bacillus anthracis exosporium
Cells from discarded dressings differentiate chronic from acute wounds in patients with Epidermolysis Bullosa
Impaired wound healing complicates a wide range of diseases and represents a major cost to healthcare systems. Here we describe the use of discarded wound dressings as a novel, cost effective, accessible, and non-invasive method of isolating viable human cells present at the site of skin wounds. By analyzing 133 discarded wound dressings from 51 patients with the inherited skin-blistering disease epidermolysis bullosa (EB), we show that large numbers of cells, often in excess of 100 million per day, continually infiltrate wound dressings. We show, that the method is able to differentiate chronic from acute wounds, identifying significant increases in granulocytes in chronic wounds, and we show that patients with the junctional form of EB have significantly more cells infiltrating their wounds compared with patients with recessive dystrophic EB. Finally, we identify subsets of granulocytes and T lymphocytes present in all wounds paving the way for single cell profiling of innate and adaptive immune cells with relevance to wound pathologies. In summary, our study delineates findings in EB that have potential relevance for all chronic wounds, and presents a method of cellular isolation that has wide reaching clinical application