Delivery of functional DNA and messenger RNA to mammalian phagocytic cells by recombinant yeast

Abstract

Among the different vaccination approaches, DNA/RNA vaccination represents a promising means in particular for the induction of effective cellular immune responses conferred by CD8-positive T lymphocytes. To achieve such immune responses, there is a need for novel delivery systems that allow the introduction of nucleic acids to the cytosol of immune cells. We show, for the first time, the delivery of functional DNA and messenger RNA (mRNA) to mammalian antigen-presenting cells, including murine macrophages and human dendritic cells, using the yeast Saccharomyces cerevisiae as the delivery vehicle. After transfer of the particular nucleic acid, subsequent antigen processing and presentation were demonstrated in a human system. Remarkably, release of DNA/mRNA does not require additional 'helper' proteins such as listeriolysin. In conclusion, the yeast-based system described here is superior to many bacterial and viral systems in terms of efficacy, safety and targeting suggesting 'mycofection' as a promising approach for the development of a novel type of live vaccines.

Intranasal mRNA nanoparticle vaccination induces prophylactic and therapeutic anti-tumor immunity

“…This is a cost, labor and resource intensive procedure. Because of these reasons, researchers have explored alternative cell-based approaches 10,11 , as well as direct in vivo injection of mRNA in naked 12,13 and nanoparticle formats [14][15][16][17][18][19] . However, due to rapid degradation of naked mRNA in vivo, direct injection of mRNA is effective only when it is injected directly into lymph nodes 12,13 .…”

Schizosaccharomyces pombe: A novel transport vehicle of functional DNA and mRNA into mammalian antigen-presenting cells

“…d118 ade6.210] [8] was grown at 30 • C in complex YEPD (2% peptone, 1% yeast extract, 2% glucose) or synthetic YE medium (5% yeast extract, 3% glucose, 0.75% uracil, 0.75% adenine). The IRES-eGFP fusion and the inducible yeast promoter ICL1 were previously described [5]. The murine DC-STAMP promoter was amplified using chromosomal DNA of IC21 cells (primers see Table 1) and routinely sequenced.…”

“…IRES-eGFP was cloned as XhoI/BglII fragment into pREP1-DC-STAMP, pREP1-ICL1, and pREP1 respectively. The yeast nmt1 terminator was replaced by the SV40 poly(A) signal through BglII/SacI restriction [5]. Yeast cells were transformed by the lithium acetate method [8] and transformants were selected on synthetic complete medium lacking leucin.…”

“…So far, mostly baker's yeast (S. cerevisiae) was used as delivery vehicle for proteinaceous antigens or, more recently, antigen-encoding DNA and mRNA to specifically activate cell-mediated immune responses [5]. DNA or mRNA vaccination has become the most prominent strategy to induce cell-mediated immunity including CD8 + T-cells [5][6][7], essential for the elimination of infected and/or tumor cells. Besides S. cerevisiae, we found that antigen-expressing nonconventional yeasts like Sz.…”

Surface-modified yeast cells: A novel eukaryotic carrier for oral application

“…In the last years, we and other groups described the successful usage of intact, recombinant yeast cells for the delivery of both proteins [17][18][19][20][21][22] and functional nucleic acids [23][24][25][26][27][28] …”