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Gewählte Publikation:

Yanez, RJR; Lamprecht, R; Granadillo, M; Torrens, I; Arcalis, E; Stoger, E; Rybicki, EP; Hitzeroth, II.
(2018): LALF(32-51)-E7, a HPV-16 therapeutic vaccine candidate, forms protein body-like structures when expressed in Nicotiana benthamiana leaves
PLANT BIOTECHNOL J. 2018; 16(2): 628-637. FullText FullText_BOKU

High-risk human papillomaviruses (HPVs) cause cervical cancer, and while there are good prophylactic vaccines on the market, these are ineffective against established infections, creating a clear need for therapeutic vaccines. The HPV E7 protein is one of the essential oncoproteins for the onset and maintenance of malignancy and is therefore an ideal therapeutic vaccine target. We fused the HPV-16 E7 protein to the Limulus polyphemus antilipopolysaccharide factor (LALF(32-51)), a small hydrophobic peptide that can penetrate cell membranes and that has immunomodulatory properties. LALF(32-51)-E7 was transiently expressed in Nicotiana benthamiana, and we previously determined that it accumulated better when targeted to chloroplasts compared to being localized in the cytoplasm. Subsequently, we aimed to prove whether LALF(32-51)-E7 was indeed associated with the chloroplasts by determining its subcellular localization. The LALF(32-51)-E7 gene was fused to one encoding enhanced GFP to generate a LG fusion protein, and localization was determined by confocal laser scanning microscopy and transmission electron microscopy (TEM). The fluorescence observed from chloroplast-targeted LG was distinctively different from that of the cytoplasmic LG. Small spherical structures resembling protein bodies (PBs) were seen that clearly localized with the chloroplasts. Larger but less abundant PB-like structures were also seen for the cytoplasmic LG. PB-like structure formation was confirmed for both LG and LALF(32-51)-E7 by TEM. LALF(32-51)-E7 was indeed targeted to the chloroplasts by the chloroplast transit peptide used in this study, and it formed aggregated PB-like structures. This study could open a new avenue for the use of LALF(32-51) as a PB-inducing peptide.
Autor/innen der BOKU Wien:
Arcalis Elsa
Stöger Eva
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Find related publications in this database (Keywords)
protein bodies
therapeutic vaccine

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