•  
  •  

Home > Health Sciences > JHDRP > Vol. 12 (2018-2019) > Iss. 4

 

Keywords

Zika Virus; Vaccines; Exosomes; Antigen-presenting vesicles

Disciplines

Medicine and Health Sciences

Abstract

With the recent emergence of Zika virus (ZIKV) diseases, increasing global concern has driven the demand for a vaccine. One promising vaccine platform has presented itself in the form of exosomes: a subgroup of extracellular vesicles released by many human cell types that facilitate intercellular communication. The objective of this study is to engineer exosomes that incorporate ZIKV structural proteins into its phospholipid bilayer. Previous studies indicate that CD9 and CD63 proteins are highly enriched in exosomal membranes. From this, it was hypothesized that attaching ZIKV genes to CD9 or CD63 to produce a gene fusion may enable exosomes to act as antigen-presenting vesicles. These engineered exosomes may potentially stimulate T-cells to mount a strong immune response. The cDNA of the CD9, CD63, and the highly immunogenic ZIKV genes (envelope, precursor membrane, and NS1) were generated using RT-PCR. These products were used as a template for regular PCR, and cloned into pcDNA3.1/V5 vector. The chimeric gene fusion was assembled using the Gibson assembly kit, and transfected into human embryonic kidney epithelial (HEK293T) cells for expression. The exosomes were purified from the supernatant and subjected to immunoblotting and immunofluorescence assays to confirm the presence of ZIKV proteins.

The results of this study are pending at the time of this abstract submission. A future study will be conducted using an in vitro activation assay to determine if the engineered exosomes induce T-cell activation. The potential candidates will be used in an animal study for immunity against ZIKV infection.

Download


Share

COinS