Surface-Layer Protein A (SlpA) is a major contributor to host-cell adherence of clostridium difficile

Michelle M. Merrigan, Anilrudh Venugopal, Jennifer L. Roxas, Farhan Anwar, Michael J. Mallozzi, Bryan A.P. Roxas, Dale N. Gerding, V. K. Viswanathan, Gayatri Vedantam

Research output: Contribution to journalArticle

54 Scopus citations

Abstract

Clostridium difficile is a leading cause of antibiotic-associated diarrhea, and a significant etiologic agent of healthcare-associated infections. The mechanisms of attachment and host colonization of C. difficile are not well defined. We hypothesize that non-toxin bacterial factors, especially those facilitating the interaction of C. difficile with the host gut, contribute to the initiation of C. difficile infection. In this work, we optimized a completely anaerobic, quantitative, epithelial-cell adherence assay for vegetative C. difficile cells, determined adherence proficiency under multiple conditions, and investigated C. difficile surface protein variation via immunological and DNA sequencing approaches focused on Surface-Layer Protein A (SlpA). In total, thirty-six epidemic-associated and non-epidemic associated C. difficile clinical isolates were tested in this study, and displayed intra- and inter-clade differences in attachment that were unrelated to toxin production. SlpA was a major contributor to bacterial adherence, and individual subunits of the protein (varying in sequence between strains) mediated host-cell attachment to different extents. Pre-treatment of host cells with crude or purified SlpA subunits, or incubation of vegetative bacteria with anti-SlpA antisera significantly reduced C. difficile attachment. SlpA-mediated adherence-interference correlated with the attachment efficiency of the strain from which the protein was derived, with maximal blockage observed when SlpA was derived from highly adherent strains. In addition, SlpA-containing preparations from a non-toxigenic strain effectively blocked adherence of a phylogenetically distant, epidemic-associated strain, and vice-versa. Taken together, these results suggest that SlpA plays a major role in C. difficile infection, and that it may represent an attractive target for interventions aimed at abrogating gut colonization by this pathogen.

Original languageEnglish (US)
Article numbere78404
JournalPloS one
Volume8
Issue number11
DOIs
StatePublished - Nov 12 2013

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    Merrigan, M. M., Venugopal, A., Roxas, J. L., Anwar, F., Mallozzi, M. J., Roxas, B. A. P., Gerding, D. N., Viswanathan, V. K., & Vedantam, G. (2013). Surface-Layer Protein A (SlpA) is a major contributor to host-cell adherence of clostridium difficile. PloS one, 8(11), [e78404]. https://doi.org/10.1371/journal.pone.0078404