Protein delivery by subviral particles of human cytomegalovirus.
Direct protein delivery is an emerging technology in vaccine development and gene therapy. We could previously show that subviral dense bodies (DB) of human cytomegalovirus (HCMV), a beta-herpesvirus, transport viral proteins into target cells by membrane fusion. Thus these non-infectious particles provide a candidate delivery system for the prophylactic and therapeutic application of proteins. Here we provide proof of principle that DB can be modified genetically. A 55 kDa fusion protein consisting of the green fluorescent protein and the neomycin phosphotransferase could be packed in and delivered into cells by recombinant DB in a functional fashion. Furthermore, transfer of protein into fibroblasts and dendritic cells by DB was efficient, leading to exogenous loading of the MHC-class I antigen presentation pathway. Thus, DB may be a promising basis for the development of novel vaccine strategies and therapeutics based on recombinant polypeptides.
Pepperl-Klindworth
S
S
Institute for Virology, Johannes Gutenberg University of Mainz, Obere Zahlbacher Strasse 67, 55101 Mainz, Germany.
Frankenberg
N
N
Riegler
S
S
Plachter
B
B
D003587:Cytomegalovirus / Q000648:ultrastructure*; D003713:Dendritic Cells / Q000378:metabolism* / Q000821:virology; D005347:Fibroblasts / Q000378:metabolism / Q000821:virology; D019085:Fluorescent Antibody Technique, Direct; D015870:Gene Expression; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D049452:Green Fluorescent Proteins; D006801:Humans; D019868:Kanamycin Kinase / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D022142:Secretory Vesicles; D019444:Vaccines, DNA / Q000008:administration & dosage*
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D049452:Green Fluorescent Proteins; D019868:Kanamycin Kinase