The choice of anchoring protein influences interaction of recombinant Bacillus spores with the immune system

  • Aurelia Piekarska
  • Paulina Pełka
  • Grażyna Peszyńska-Sularz
  • Alessandro Negri
  • Krzysztof Hinc
  • Michał Obuchowski
  • Adam Iwanicki Intercollegiate Faculty of Biotechnology UG-MUG Department of Medical Biotechnology Medical University of Gdańsk
Keywords: Bacillus subtilis, spore display, APCs, FliD, Clostridium difficile, immune response


The technology of display of heterologous proteins on the surface of Bacillus subtilis spores enables use of these structures as carriers of antigens for mucosal vaccination. Currently there are no technical possibilities to predict, whether a designed fusion will be efficiently displayed on the spore surface and how such recombinant spores will interact with cells of the immune system. In this study we compared four variants of B. subtilis spores presenting a fragment of FliD protein of Clostridium difficile in fusion with CotB, CotC, CotG or CotZ spore coat proteins. We show that these spores promote their phagocytosis and activate both, J774 macrophages and JAWSII dendritic cells of murine cell lines. Moreover, we used these spores for mucosal immunization of mice. We conclude that observed effects vary with the type of displayed FliD-spore coat protein fusion and seem to be mostly independent on its abundance and localization in the spore coat structure.


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