JoVE Encyclopedia of Experiments
Immunology
0 views • 5:35 min • July 8th, 2025
Take a microplate with serial dilutions of a serum sample containing antibodies against the Zika virus.
Add Dengue reporter virus particles or RVPs, and incubate.
The engineered RVPs comprise an envelope with structural proteins and a nucleocapsid containing genomic RNA, encoding a fluorescent protein.
The structural homology of Dengue and Zika viruses' envelope proteins causes the antibodies to cross-react with the RVPs, forming a complex.
Add human leukemia cells impermissible to infection by RVPs.
The cells possess Fc receptors that recognize the RVP-bound antibodies, facilitating RVPs' entry inside endosomes — termed antibody-dependent enhancement or ADE.
Pellet the cells and discard the supernatant containing non-internalized RVPs. Add a growth media and incubate.
The pH-dependent fusion of RVP and endosomal membranes releases the nucleocapsid, which disassembles to release genomic RNA, enabling fluorescent protein expression.
Fix the cells and analyze them using flow cytometry.
A higher fluorescence at a suitable antibody concentration represents increased RVP internalization, indicating enhanced infection.
Begin by thawing the serum samples at room temperature. When thawed,
View the full transcript and gain access to thousands of scientific videos