Nanokosmokseen saavuttu ja uutta rokotekehittelyä mahdollistuu exosomitasolta

https://www.ncbi.nlm.nih.gov/pubmed/29274250

Biotechnol J. 2018 Apr;13(4):e1700443. doi: 10.1002/biot.201700443. Epub 2018 Mar 24.

An Exosome-Based Vaccine Platform Imparts Cytotoxic T Lymphocyte Immunity Against Viral Antigens.Anticoli S¹, Manfredi F¹, Chiozzini C¹, Arenaccio C¹, Olivetta E¹, Ferrantelli F¹, Capocefalo A², Falcone E², Ruggieri A², Federico M¹. Abstract

Exosomes are 50-150 nm sized nanovesicles released by all eukaryotic cells. The authors very recently described a method to engineer exosomes in vivo with the E7 protein of Human Papilloma Virus (HPV). This technique consists in the intramuscular injection of a DNA vector expressing HPV-E7 fused at the C-terminus of an exosome-anchoring protein, that is, Nef^mut , the authors previously characterized for its high levels of incorporation in exosomes. In this configuration, the ≈11 kDa E7 protein elicited a both strong and effective antigen-specific cytotoxic T lymphocyte (CTL) immunity. Attempting to establish whether this method could have general applicability, the authors expanded the immunogenicity studies toward an array of viral products of various origin and size including Ebola Virus VP24, VP40 and NP, Influenza Virus NP, Crimean-Congo Hemorrhagic Fever NP, West Nile Virus NS3, and Hepatitis C Virus NS3. All antigens appeared stable upon fusion with Nef^mut , and are uploaded in exosomes at levels comparable to Nef^mut . When injected in mice, DNA vectors expressing the diverse fusion products elicited a well detectable antigen-specific CD8⁺ T cell response associating with a cytotoxic activity potent enough to kill peptide-loaded and/or antigen-expressing syngeneic cells. These data definitely proven both effectiveness and flexibility of this innovative CTL vaccine platform. KEYWORDS:

CTL immunity; DNA immunization; Ebola virus; HIV-1 Nef; exosomes


(2) 
https://www.ncbi.nlm.nih.gov/pubmed/30167966 

Mol Biotechnol. 2018 Nov;60(11):773-782. doi: 10.1007/s12033-018-0114-3.

DNA Vectors Generating Engineered Exosomes Potential CTL Vaccine Candidates Against AIDS, Hepatitis B, and Tumors. Ferrantelli F¹, Manfredi F¹, Chiozzini C¹, Anticoli S¹, Olivetta E¹, Arenaccio C¹, Federico M². Abstract

Eukaryotic cells constitutively produce nanovesicles of 50-150 nm of diameter, referred to as exosomes, upon release of the contents of multivesicular bodies (MVBs). We recently characterized a novel, exosome-based way to induce cytotoxic T lymphocyte (CTL) immunization against full-length antigens. It is based on DNA vectors expressing products of fusion between the exosome-anchoring protein Nef mutant (Nef^mut) with the antigen of interest. The strong efficiency of Nef^mut to accumulate in MVBs results in the production of exosomes incorporating huge amounts of the desired antigen. When translated in animals, the injection of Nef^mut-based DNA vectors generates engineered exosomes whose internalization in antigen-presenting cells induces cross-priming and antigen-specific CTL immunity. Here, we describe the molecular strategies we followed to produce DNA vectors aimed at generating immunogenic exosomes potentially useful to elicit a CTL immune response against antigens expressed by the etiologic agents of major chronic viral infections, i.e., HIV-1, HBV, and the novel tumor-associated antigen HOXB7. Unique methods intended to counteract intrinsic RNA instability and nuclear localization of the antigens have been developed. The success we met with the production of these engineered exosomes opens the way towards pre-clinic experimentations devoted to the optimization of new vaccine candidates against major infectious and tumor pathologies. KEYWORDS:

Constitutive transport elements; Exosomes; HBV; HIV-1; HOXB7; Nef

PMID:

30167966

DOI:

10.1007/s12033-018-0114-3

[Indexed for MEDLINE]