Antibody-Dependent Enhancement of Zika Virus by Dengue Antibodies in Human Primary Macrophages

Begin with serial dilutions of human serum containing dengue virus antibodies.
Add the thawed Zika virus to the diluted serum and incubate.
The Zika and dengue envelope proteins share a similar structure. This allows dengue antibodies to bind to the envelope proteins of the Zika virus and form immune complexes.
Take a culture of human primary macrophages expressing Fcγ receptors.
Remove the culture medium and wash the cells with buffer.
Then, introduce the immune complexes and incubate. The Fcγ receptor recognizes the antibody-bound virus and internalizes it through endocytosis, a process termed antibody-dependent enhancement.
Following incubation, wash to eliminate unbound immune complexes.
Add fresh nutrient-rich medium and incubate.
Inside the cell, the Zika virus replicates its RNA and produces viral proteins, leading to progeny assembly within the cytoplasm.
The macrophages with enhanced Zika virus infection are now ready for downstream analysis.

Begin by seeding three times ten to the fourth cells in 100 microliters of cell culture medium per well in a sterile, flat bottom 96-well plate. When all the cells have been plated, place the plate in a 37 degree Celsius and 5% carbon dioxide i